Within the frame of this article, briefly but comprehensively, we present the existing knowledge, perspectives, and challenges for the utilization of Layered Double Hydroxides (LDHs) as adsorbents against a plethora of pollutants in aquatic matrixes. The use of LDHs as adsorbents was established by considering their significant physicochemical features, including their textural, structural, morphological, and chemical composition, as well as their method of synthesis, followed by their advantages and disadvantages as remediation media. The utilization of LDHs towards the adsorptive removal of dyes, metals, oxyanions, and emerging pollutants is critically reviewed, while all the reported kinds of interactions that gather the removal are collectively presented. Finally, future perspectives on the topic are discussed. It is expected that this discussion will encourage researchers in the area to seek new ideas for the design, development, and applications of novel LDHs-based nanomaterials as selective adsorbents, and hence to further explore the potential of their utilization also for analytic approaches to detect and monitor various pollutants. Freshwater quality has been threatened over the few last decades due to rapid urbanization. To meet the enhanced demand for products, the expansion of industrial and agricultural activities is taking place at an accelerated pace, which ends up releasing a large number of contaminants into water bodies ,2. In tuDyes: Various colored substances are widely utilized as dyes by the textile, food, paper, and pharmaceutical industries. Among these sectors, the textile industry releases 10 to 15% of the dyes produced in the environment. These elements, even in low concentrations, can pose a threat to the environment and living beings. The presence of dyes in water resources reduces the penetration of light through the water, affecting photosynthesis and dissolved oxygen levels, and harming the aquatic biota. In addition, some dyes can degrade to produce highly toxic and carcinogenic compounds ,4,5.Metals: Effluents from industries such as battery manufacturing, electroplating, mining, smelting, and many other industrial activities consist of heavy metals mixtures . The presence of these metals in water bodies is harmful to human health, as they are bioaccumulative and toxic. Among the negative effects of these elements in living beings, even at a low concentration are diarrhea, stomach pain, headaches, chronic bronchitis, and lung cancer ,6,7.Pharmaceuticals: Drugs are widely used in human medicines, livestock, aquaculture, beekeeping, and poultry to aid species growth. Currently, the release of these elements into the environment is not always absolutely regulated/controlled; hence, it is difficult to monitor their effects on living beings. The discharge of drugs into water resources occurs through the excretion of compounds not metabolized by living beings and industrial and hospital effluents. The continued presence of these pharmaceutical pollutants can cause oxidative stress and negative effects on reproduction , osmoregulation, and altered immune functions in aquatic biotics ,9.Emerging pollutants: In general, emerging pollutants are chemical or microbiological products that are not commonly monitored or regulated and can cause problems to the environment as well as to the health of living beings. Except for the abovementioned, substances that can end up acting as pollutants are usually found in personal care products, pharmaceuticals, industrial additives, pesticides, plasticizers, and solvents. Among these substances, exposure to emerging organic compounds such as phenol, benzene, toluene, and xylenes can cause serious health problems such as gastrointestinal disorders, lung and kidney damage, heart attacks, and cancer. Yet, the increased use of pesticides and personal care products has increased dramatically in recent years. Although these products appear in low concentrations in water bodies, they can negatively influence the environment due to their resistance to biodegradation and accumulation in tissues ,10.Among the several technologies applied for water treatment, adsorption using layered double hydroxides (LDHs) will be highlighted in this article. Adsorption is a unitary operation in which the separation process occurs due to contact between a fluid phase—in this case, liquid—containing one or more contaminants (adsorbate) to be adsorbed/removed, and a solid (adsorbent). Due to the imbalance of forces (attraction or repulsion) for adsorption, the contaminant is attracted to the solid surface by physical or chemical interactions. This mass transfer process occurs until the balance between the adsorbed contaminant and what remains in the liquid phase, the residual, is reached. This is termed equilibrium. Adsorption is recognized as an effective process for water treatment due to certain characteristics, such as high efficiency, feasibility, low cost, flexibility, simplicity, wide processing range, cost-effective applications for water treatment, easy operation and implementation, great availability, and the possibility of adsorbents’ regeneration. A very important aspect is that the adsorption-based processes do not result in the formation of hazardous substances/by-products ,11,12,13Around 70% of the adsorption operation cost is related to the adsorbents . TherefoCurrently, and within this context, layered double hydroxides (LDHs), also known as anionic clays, have drawn great attention in their application as adsorbents. Several researchers have proposed the use of LDHs as efficient adsorbents against a plethora of organic and inorganic contaminants ,16,17,18(1)Physical adsorption. LDHs can have a high specific surface area, and hence present high adsorption capacities due to the presence and availability of active adsorption sites. Furthermore, the specific surface area of LDHs can be increased through calcinating or modifying/depositing on supports with three-dimensional structures.(2)Ion exchange. Strongly negative molecules can be easily changed for the original anions in LDHs. In addition, positive ions can also be exchanged in the intermediate layer of LDHs, if pre-interleaved by some chelators.(3)Interleaving. This starts from a preparation process, such as co-precipitation. Furthermore, the capture of molecules via the intercalation process is faster and more complete than ion exchange.Considering the enhanced surface chemical heterogeneity and the nanostructured nature of LDHs, their elevated remediation efficiency against different substances is linked in the literature to plenty of physical and/or chemical interactions, with the most reported ones collected in For the design and utilization of a proper adsorbent, certain characteristics are essential, such as low production cost, thermal, mechanical, and chemical stabilities, desirable physicochemical characteristics , high efficiency and adsorption capacity, rapid kinetics, and regeneration/reuse potential . Some oflamellae 2]x+. Am−x/m.nH2O [2+ represents a divalent metal cation; M3+, the trivalent cation; Am−, an anion intercalated with charge m; X, the ratio between divalent (M2+) and trivalent cations (M3+), for which the value of the molar ratio can be between 2.0 and 4.0; and n, the number of soft water molecules. Many divalent  and trivalent  metal cations can be used for the preparation of LDHs. Except for bivalent and trivalent metal ions, a wide range of monovalent and tetravalent metal ions  may also be inserted in the octahedral sites [−, Cl−, Br−) in the interlamellar space [LDHs are two-dimensional (2D) nanostructures composed of stacked layers consisting of mixed hydroxides of di- and trivalent cations with hydrated anions in the spaces between the positively charged lamellae . LDHs arnosheets . In genex/m.nH2O , where Mal sites . In addiar space . Because(i)Direct methods: The preparation of LDHs occurs via direct precipitation from the addition of tri- and divalent cations, in a solution in alkaline pH with the main methods of coprecipitation, salt–oxide, sol–gel, induced hydrolysis, and hydrothermal synthesis.(ii)Indirect methods: involve replacing an interlamellar anion from a previously produced precursor LDH. Examples of this substitution method are ion exchange in solution, ion exchange in acidic medium, double phase replacement, and regeneration through the delaminate precursor ,36,37. TFor LDHs, the precisely controlled chemical composition of the lamellar layers and the interlayer composition provide a unique supramolecular nanometric structure with the ability to disperse active sites on an atomic scale, in addition to facilitating morphological manipulation ,34. ThusAs already mentioned, LDHs are very promising materials for the removal of a wide variety of pollutants via adsorption. There are many reports in the literature; consequently, we will address some of the most important ones that gather the highest level of research attention for specific pollutants, with emphasis on the utilized LDHs.Dyes: The presence of dyes in water and wastewater is highly undesirable, and LDHs have been reported as efficient remediation media of a large number of industrial dyes . Dyes ar3), through the coprecipitation method to remove methyl orange from aqueous solution. The prepared LDHs showed the intercalary-like structure with (00l) reflections and a nanoscale nature, as well as the presence of the desired anions in the interlayer space. The maximum capacities were 185.06 mg·g−1 for Mg-Al-Ds and 97.5 mg·g−1 for Mg-Al-CO3. They also observed the influence of the pH of the solution on the adsorption capacity of the dye in the case of Mg-Al-CO3. The adsorptive capacity of Mg-Al-Ds was not influenced by the pH of the solution, while Mg-Al-CO3 showed greater efficiency at pH in the range of 3–7. According to the authors, this behavior may be associated with two mechanisms: (1) anion exchange of carbonate anions for dye anions; (2) association of the positively charged surface groups of LDHs and dye anions.El-Abboubi et al.  evaluate3-LDH double hydroxides using the coprecipitation method for the removal of dye RB19 from an aqueous solution. The material presented a surface area of around 48 m2·g−1, high crystallinity and the presence of carbonate, hydroxyl, and metal ions groups within its structure. The proposed adsorption mechanisms involved in the process were electrostatic attraction, physisorption, and chemical bonding. The maximum capacity was found to be equal to 367.93 mg·g−1.Kostić et al.  synthesiAhmed et al.  prepared2+, Cu2+ and Zn2+) [Metals: The discharge of heavy metals into water bodies results in harmful effects to the environment and human health due to their toxicity and persistence. The presence of these elements in water systems has caused concern in recent years; hence, the interest in novel and efficient remediation media is still elevated ,16. In tnd Zn2+) . There ind Zn2+) ,48.2+ ions from an aqueous solution. The results showed that NC10% showed higher adsorptive capacity  compared to NC5%  and NIBP , demonstrating the favoring of adsorption in the presence of Ca/Fe double hydroxides.Dinari and Neamati  synthesi6+ from an aqueous solution. The material presented a surface area of 354 m2·g−1, and the SEM analysis showed an irregular block structure. In this study, it was proposed that predominantly electrostatic interaction between the metal ion and the adsorbent took place with a maximum capacity of 28.2 mg·g−1 (0.54 mmol·g−1). The results demonstrate potential application prospects in the removal of Cr6+ in wastewater.Guo et al.  synthesi2+, Pb2+, and Cu2+ from an aqueous solution. The characterization results showed characteristic peaks at 2θ = 13.24°, 22.88°, 35.09°, 39.19°, 47.23°, and 60.97°, while CO, COO− and CH surface functional groups were also detected in its structure. The maximum adsorption capacities of SA-LDH were 60 mg·g−1 (0.945 mmol·g−1) for Cu2+, 243.66 mg·g−1 (1.176 mmol·g−1) for Pb2+, and 95.55 mg·g−1 (0.850 mmol·g−1) for Cd2+. The results showed that the possible mechanisms involved in the adsorption process are: (1) bonding or complexation with SurOH or Sur-O- of SA-LDH; (2) precipitation of metal hydroxides or carbonates; (3) isomorphic substitution, and (4) chelation with COO− in the interlayers.Zhang et al.  synthesiOxyanions: High levels of oxyanions found in the environment are reported all over the world. Oxyanions  are considered dangerous to humans and wildlife, even at very low concentrations ,49. In t−1 for Zr-LDO.Motandi et al.  synthesi−1 and 33.21 mg·g−1 for arsenate and phosphate, respectively. For the multicomponent system, the maximum adsorption capacities were 16.22 mg·g−1 for arsenate and 20.26 mg·g−1 for phosphate. In the single-component system, a possible adsorption mechanism is the ion exchange between the nitrate interlayer and the arsenate or phosphate groupings. In the multicomponent system, coordinated bonds are also possibly responsible for the competition of arsenate or phosphate in the adsorptive process.Jung et al.  synthesi2·g−1; the presence of –OH, N=O, M–OH and M–O–M was detected in its structure. The maximum amount of nitrate adsorption was 10.56 mg·g−1. The adsorptive process was spontaneous and exothermic. The possible proposed adsorption mechanisms of nitrate removal using FeMgMn-LDH were an electrostatic attraction and ion exchange.Zhou et al.  prepared−1 or µL−1) are assumed to be emerging pollutants nowadays. These components are used in different industrial processes for the production of pharmaceuticals, personal care products (PCPs), beverages, foods, and others [Emerging pollutants: Plenty of synthetic substances that have recently been detected at low concentrations Santamaría et al.  synthesi2O3 as an adsorbent for the removal of diclofenac from an aqueous solution. The material presented the main diffraction peaks 003, 006, 012, 041, 111, 241, and 110, and a surface area of 102 m2·g−1. The Zn-Al-LDH.xBi2O3 presented a capacity of 574.71 mg·g−1 for the removal of diclofenac. The results indicated that the adsorptive process occurs in a monolayer and that the diffusion of diclofenac occurs mainly on the external surface.Kumari et al.  used a dOther kinds of pollutants: There are few reports of the use of LDHs for the adsorptive removal of bacteria and viruses, per- and poly-fluoroalkyl substances, and rare earth and radioactive substances; such as Cs, Sr, and Th . Therefore, exploring the use of LDHs in the removal of this type of contaminant is very promising and important for environmental health protection.Utilization in real-life applications: Industrial effluents contain various pollutants simultaneously, such as dyes, metals, pesticides, and antibiotics, which impose a high physicochemical complexity on the real system. Therefore, it is necessary to understand the interaction between the adsorbates and the adsorbent in the water decontamination process ,3. DespiConsidering the adsorption mode, batch experiments are well reported in the literature. On the contrary, although the design and efficiency are well approached in lab-scale batch studies, the fixed beds procedure has received little attention regarding exploration of the use of LDHs as adsorbents. The fixed bed columns are applied for continuous flow, and are effective for the treatment of large volumes of effluents, allowing application in industries, adsorption–desorption cycles, efficient fluid–particle contact and easy phase separation, which makes the treatment process cheaper and more sustainable ,55,56. IFixed beds are employed primarily for commercial water treatment scale-up. Discovering and proposing new operational modes to improve fluid–particle interaction and phase separation is a key issue. Therefore, a smaller treatment area, faster operation, larger water volumes treated, and cost reduction are important advantages. As a result, there may be benefits such as a smaller treatment area, faster operation, treatment of bigger quantities of water, and cheaper costs. Therein, the study of alternative contacting devices, such as fluidized bed, spouted bed, simulated moving bed, expanded bed, continuous stirred tanks, and multi-batch tanks, is much appreciated and warrants substantially further investigation .PZC), Fourier transform infrared spectroscopy (FT-IR), and X-ray diffraction (XRD) helps us to understand the adsorptive mechanism of the process. The most widely proposed interactions/mechanisms involved in the adsorption of organic compounds generally involve electrostatic interaction between the surface of the LDH (positively or negatively charged) with the cationic (positively charged) or anionic (negatively charged) compounds. The adsorptive mechanism associated with the adsorption of metal ions involves the ion exchange between the surface of the material and the metal ions. Other mechanisms are also involved in the adsorption of metal ions in LDHs, such as precipitation and complexation. Among the functional groups present in the structure of LDHs, the hydroxyl group favors the adsorption of both organic and inorganic compounds. Another important parameter in the adsorption process using LDHs is the pH of the solution. It is observed that, in general, the favorable pH for the adsorption of anionic compounds occurs at pH lower than 7 and for cationic compounds at pH above 7.The studies presented and discussed above demonstrate the prosperity of utilizing LDHs as remediation media, and hence, the growing interest of researchers against various pollutants regarding interpretation and proposals for the involved mechanisms and interactions. The studies revealed elevated adsorptive capacities of organic and inorganic pollutants, even compared to benchmark materials such as porous carbons ,16,45,48Layered double hydroxides (LDHs) represent one category of materials with an increasing trend of interest toward the removal of water pollutants. To uplift the direction of LDHs-based adsorbent application on a commercial scale, novel approaches and considerations with emphasis on sustainability and low cost are presented. However, more adsorption investigations of LDHs need to be explored in multi-component systems rather than in the current trend, where most of the studies are limited to mono-component systems and lab-scale batch experiments. The parametric influence on the adsorption process requires an advanced optimization approach to attain maximum removal performance of water pollutants using LDHs-based adsorbents. The major setback of a “one time” use of the adsorbent must be overcome with the proper assessment of desorption strategies, followed by a good number of adsorbent recycle options, treated water re-use, and the adsorbate pollutant recovery and re-use. More mechanistic insights into water pollutant removal need to be understood in multicomponent simulated and real effluent systems to study the antagonistic effects targeting the development of adsorbents with high selectivity for the specific pollutant(s). To understand the complete application of layered double hydroxides in water pollutant elimination, the batch system followed by a continuous system needs to be performed at lab and pilot scale, with real effluents.The simplicity and low cost of LDHs production , in addition to the ability to tailor design their composition and structure, establish them as sustainable and attractive candidates for real-life applications. Moving a step forward regarding the advantages of nanomaterials, designing and synthesizing novel nanocomposites/hybrids should be assumed as a worthwhile effort to be further explored in the field of research. The development of synthesis methods capable of producing LDHs on a commercial scale is one of the current problems for the use of these materials on an industrial scale. In addition, LDHs syntheses are generally limited to the use of MgAl and MgFe, requiring the exploration of new compounds. Within the frame of circular (bio)economy and sustainability, the development of novel and advanced composited of LDHs with biomass-derived biochar/carbon and determining the optimum ratio between the counterparts is a prosperous topic of research, since still only a few studies exist.


Recovery after anesthesia has a significant impact on a patient’s return to daily life. This study was performed to compare the postoperative quality of recovery according to the method of anesthesia administered among patients undergoing OPCAB using the Korean version of the Quality of Recovery-40 (QoR-40K) questionnaire. This single-blind, prospective study  was performed using a population of 102 patients undergoing OPCAB under general anesthesia. The patients were randomly assigned to one of two groups using a computer-generated list: a total intravenous anesthesia group (Group T) and a balanced anesthesia group (Group B). The QoR-40K score was measured preoperatively and at 24 and 48 h after extubation. There was no significant difference in the QoR-40K scores between the groups at 24 and 48 h after extubation. In addition, there were no significant differences between groups with respect to any of the five dimensions of QoR-40K at 24 and 48 h after extubation. Finally, there were no differences in the postoperative opioid consumption, time to extubation, or length of hospital stay. In this study, there was no difference in the QoR-40K score at 24 h after extubation between Groups T and B. Therefore, both methods of anesthesia are suitable for use when performing OPCAB. Advances in anesthesia and surgical procedures have been accompanied by decreased rates of perioperative mortality and morbidity, and the focus of anesthesia management is now shifting to the quality of recovery after surgery . The quaCoronary artery bypass grafting is a widely performed surgical method involving the bypassing of a partially or completely blocked coronary artery using an autologous artery or vein as a graft . CoronarPoor-quality recovery after cardiac surgery leads to a decline in quality of life during the 3 months following surgery . HoweverThis prospective, randomized, single-blind, controlled study was performed between March 2020 and August 2021 in single tertiary care hospital. The study protocol was approved by the Institutional Review Board of Chonnam National University Hospital , and the study was registered at the Clinical Research Information Service of the Korea National Institute of Health . All participants provided written informed consent before enrollment in accordance with the principles of the Declaration of Helsinki. Patients aged 20–79 years, corresponding to American Society of Anesthesiologists (ASA) physical status I–IV, and who were undergoing off-pump coronary artery bypass graft surgery (OPCAB) were enrolled in the study. Patients who were taking any sedatives, opioids, or sleep aids and those with a history of allergic reactions to any of the study drugs were excluded. Patients with physical disabilities, those who were illiterate, those who underwent conversion to coronary artery bypass grafting ), those with severe left ventricular dysfunction (ejection fraction < 30%), those who suffered from major organ failure before surgery , or who required continuous mechanical ventilation assistance after surgery were also excluded.Pre-anesthesia visits and evaluations were conducted on the day before, or day of, surgery to confirm the patient’s general condition, preoperative test results, and medications. In addition, demographic information, height, weight, comorbidities, EuroSCORE II, and the number of affected coronary vessels were extracted from the patients’ medical records. At this time, a blinded investigator used the QoR-40K questionnaire to obtain baseline values for all patients. The patients were divided into two groups according to anesthesia method, i.e., a total intravenous anesthesia group (Group T) and balanced anesthesia group (Group B). The group assignment process was performed based on a computer-generated list of random numbers by a researcher who did not participate in the administration of anesthesia. Researchers investigating the QOR-questionnaire were unaware of the assignment.After entering the operating room, each patient was monitored by 5-lead electrocardiography, pulse oximetry, and noninvasive blood-pressure monitoring. Before inducing anesthesia, we implanted a 22 or 20 G catheter into the radial artery and began continuous arterial blood-pressure monitoring. After recording the baseline vital signs, 8 L/min of oxygen was applied via a face mask and sufentanil (0.3 μg/kg) was infused. ®; Fresenius Kabi Korea Ltd., Seoul, Korea) with the target plasma concentration set to 1.5–2.0 μg/mL. Infusion was increased in increments of 0.5 μg/mL until reaching a target bispectral index (BIS) of 40–60. In Group B, a bolus (1–1.5 mg/kg) of propofol  was administered slowly until the target BIS was reached. Sufentanil infusion was started at the beginning of anesthesia and was then infused continuously with a mean arterial pressure (MAP) of 70–90 mmHg in both groups. After loss of consciousness was confirmed, rocuronium (0.8–1.0 mg/kg) was administered intravenously to facilitate intubation. During maintenance of anesthesia, propofol or sevoflurane administration was adjusted according to the target BIS of 40–60. Fluids and vasopressors were used if necessary. In addition, capnography parameters, pulmonary artery pressure, transesophageal echocardiography parameters, and body temperature were monitored for all patients.Group T received target-controlled infusion of propofol . During surgery, body temperature was maintained within the normal range using an external heated mattress. Anesthetic management information, such as surgery time, anesthesia time, sufentanil dose, infused fluid volume, transfusion units, inotropic drug usage, and number of anastomotic vessels, was recorded. Blood pressure and heart rate (HR) were recorded at each time point after anesthesia induction, skin incision, sternotomy, anastomosis, protamine injection, and arrival at the intensive care unit (ICU) or general ward. After the operation, 0.075 mg of palonosetron was administered to all patients and they were transferred to the surgical ICU while maintaining mechanical ventilation. Extubation was performed by the attending physician if the patient met the criteria for extubation . Postoperative pain control was achieved using an intravenous patient-controlled analgesia (PCA) device . Tramadol (50 mg) was administered intravenously as an additional analgesic if the patient complained of pain  score ≥ 4). Postoperative nausea and vomiting (PONV) scores were recorded . Rescue antiemetics were injected in patients with PONV scores ≥ 3.The primary outcome was the QoR-40K score 24 h after extubation. Secondary outcomes were the QoR-40K score 48 h after extubation; peri-operative vital signs (MAP and HR); total amounts of sufentanil, fluid, and blood products administered during surgery; use of inotropics; time to extubation after surgery; length of stay in the ICU; and length of stay in the hospital. Pain score (NRS), use of rescue analgesics, post-operative fentanyl consumption, post-operative nausea and vomiting (PONV), and use of rescue antiemetics were also measured as secondary outcomes at the time of extubation and 24 and 48 h thereafter. All researchers in this study were anesthesiologists.The patient’s HR and blood pressure at the time of extubation and 24 and 48 h thereafter, pain at the surgical site, the amount of fentanyl used in the pain-control device, the use of additional analgesics, and the use of antiemetics were determined from the medical records. The number of transplanted vessels was confirmed based on the operation records. At 24 and 48 h after extubation, the questionnaires were completed by the participants themselves or by responding to what the researcher read to them.A difference of 10 points in the QoR-40K score is associated with a 15% improvement in the quality of recovery . The sizp < 0.05 was taken to indicate statistical significance.Data were analyzed using SPSS 23.0 . The Kolmogorov–Smirnov test was performed to examine the normality of the data. Continuous variables with a normal distribution were compared using the two-sample independent t test, and continuous variables that were not normally distributed were compared using the Mann–Whitney test. Categorical data were compared using Fisher’s exact test. In all analyses, A total of 106 patients were assessed for eligibility, and 102 gave informed consent and were randomly assigned to the study groups. In Group T, four patients were excluded because of conversion to on-pump CABG (n = 1), reintubation after extubation (n = 1), or the application of extracorporeal membrane oxygenation (ECMO) (n = 2) after surgery. In Group B, three patients were excluded due to conversion to on-pump CABG (n = 2) and the application of ECMO after surgery (n = 1). Statistical analyses were performed with data from 48 patients in Group B and 47 in Group T . There were no significant differences in the demographic characteristics, comorbidities, or medications between the two groups , or in tp = 0.102). There was no significant difference in the QoR-40K score at 24 h after extubation between Groups B and T . There were no significant differences between the two groups in the scores for the five dimensions of the QoR-40K at 24 h or 48 h after extubation (p = 0.285). In addition, there was no significant difference in terms of post-operative pain  or length of hospital stay . There were no serious complications, including death, after surgery in either group.The HR was significantly lower in Group T than Group B at the time of skin incision and sternotomy . The MAPIn the present study, there were no difference in the QoR-40K scores at 24 h after extubation according to the method of anesthesia administered to patients undergoing OPCAB surgery, which was the primary outcome. In addition, there were no significant differences between the two groups in terms of the QoR-40K score at 48 h after extubation and with respect to any of the five dimensions of the QoR-40K at 24 or 48 h after extubation.Patient satisfaction after surgery is not only related to the absence of post-operative complications or the maintenance of stable vital signs but also to functional recovery, such as subjective symptoms, psychological satisfaction, and physical independence. Therefore, a multifaceted approach is required ,15,16. TSeveral clinical studies showed differing results regarding the quality of recovery according to the method of anesthesia administered for various surgeries. In endoscopic sinus surgery, the QoR-40 score at 6 h after surgery was significantly higher in a total intravenous anesthesia (using propofol) group compared with a desflurane group. In thyroid surgery, total intravenous anesthesia using propofol improved the overall recovery quality score by reducing PONV ,18. HoweIn this study, there was no difference in the QoR-40K scores between the two groups. In previous studies comparing QoR-40 scores between patients treated with general anesthesia using total anesthesia and inhalational anesthesia, the dimensions showing group differences were physical comfort, including PONV, and physical independence ,20. In tp = 0.186) at the time of extubation, 4.6 and 4.4, (p = 0.540) at 24 h after extubation, and 3.0 and 3.1 (p = 0.935) at 48 h after extubation, respectively. There were no differences in PCA dose or rescue analgesic administration between the two groups. All patients underwent surgery performed by one surgeon and the same surgical team, and there was no difference in the amount of sufentanil administered during surgery between the two groups. Furthermore, there were no differences between the two groups in terms of the pain and total QoR-40K scores.Pain is an important factor in patient satisfaction after surgery. In a previous study, preoperative administration of pregabalin to patients undergoing OPCAB surgery reduced the postoperative analgesic dose and pain score, which improved recovery scores . In thisThere were no differences between the two groups regarding the secondary outcomes of this study, i.e., the MAP during and after surgery, the dose of sufentanil administered during surgery, the use of inotropics, the time to extubation after surgery, the length of stay in the ICU, or the length of stay in hospital. There were also no differences between the two groups regarding the rate of conversion to on-pump during surgery or ECMO after surgery. This study had several limitations. First, objective laboratory data such as myocardial enzyme levels could not be collected; thus, differences between the two groups could not be determined. Second, the study was conducted at a single institution and predominantly incorporated males . Therefore, it is difficult to generalize the results of this study. Finally, as we included patients with left ventricular function (ejection fraction) ≥ 30%, we could not confirm the results in high-risk patients with severe left ventricular failure.In this study, there were no differences in the QoR-40K score at 24 h after extubation between the patients undergoing OPCAB surgery with total intravenous anesthesia and balanced anesthesia using sevoflurane. In addition, there were no differences between the two groups regarding hemodynamic parameters except heart rate during surgery, nor were there any differences concerning the length of stay in the ICU or hospital.


The aim of this meta-analysis was to systematically review the relationships between lactation diet and the composition of mature breast milk through screening multiple databases and gray literatures, with priority given to quantitative articles published in Chinese and English. We identified 27 cross-sectional studies that included 4355 lactating women. According to these data, dietary patterns were categorized into four patterns based on the fat and protein energy supplying ratio, including rational-fat and rational-protein dietary (RR), high-fat and high-protein dietary (HH), rational-fat and high-protein dietary (RH), and high-fat and rational-protein dietary (HR). The results showed the fat content in mature milk was increased with increments of fat intake, while both the protein and lactose contents in mature milk did not increase with their intakes for lactating mothers. Among these four dietary patterns, the energy ratio of macronutrients in human milk at the RR was the closest to the estimated energy ratio for infants aged 0–6 M. In conclusion, our study represents that the rational dietary pattern should be advocated, and the irrational dietary patterns, especially high-fat and high-protein dietary patterns, should be avoided during lactation stage. Optimal nutrition for lactating mothers (LMs) is critical to both the mother’s health and the infant’s short-term and even long-term growth and development. The reasonable intakes of macronutrients, including protein, fat, and carbohydrates, are associated with a decreased incidence of infections and chronic diseases for infants, such as gastrointestinal infections, autoimmune diseases, and metabolic syndrome ,2. Some diseases ,9,10. Sodiseases ,12,13. Ndiseases . When thdiseases ,16,17. Wdiseases ,19. All diseases ,15,20,21There are many factors affecting the content of breast milk, including maternal factors , infant factors (birth order and gender), and physiological factors, among which the maternal diet is only one factor that can be interfered ,22,23. SIn published reports, while cross-sectional studies were mainly used in investigating and analyzing the effect of dietary intake on the breast milk composition of LMs, its shortcomings were that they could not provide evidence to prove the temporal relationship among different variables ,35,36. TThis review aimed to explore how different maternal dietary patterns affected the macronutrients in breast milk by assessing the values and heterogeneous sources of breast milk macronutrients. A holistic perspective was used to assess the associations between the macronutrients in breast milk and dietary patterns. All the observational studies and interventional studies, classic reviews, and systematic reviews were pooled and reviewed in our study to achieve these objectives. Then, four dietary patterns were defined based on the energy ratios of protein and fat intake in total energy, and the associations between the macronutrients of breast milk and dietary patterns were finally drawn by meta-analyses.The results of a systematic review were graphically summarized to show how breast milk composition changes with diet patterns. Since breast milk macronutrients varied greatly with lactation stages, only the values of macronutrients from 1–3 M of mature breast milk were collected for the meta-analyses to decrease the systematic error and obtain the accurate estimates.https://www.webofscience.com/wos/alldb/basic-search, accessed on 23 June 2022), PubMed , the China National Knowledge Infrastructure (CNKI) , and WanFang . The following search strategy was applied: “(human OR breast) AND (milk) AND (dietary intake) AND (composition OR concentration OR content OR quantification OR amount)”. A snowball method, through manual cross-references from retrieved articles, was conducted to identify any additional relevant publications.This review was conducted in accordance with the Preferred Reporting Items for Systematic Review and Meta-Analysis (PRISMA) Guidelines . The litThe inclusion and exclusion criteria are listed in the PICOS table . QualityThe following information was collected: (1) the first author, publication year, age, body mass index (BMI) of mothers, regions, and sample size of the study; (2) four dietary patterns; (3) the outcome index, mean values, and standard deviation (SD) of macronutrients in the breast milk of the LMs; and (4) the energy ratio of macronutrients in the total energy. Dietary energy mainly comes from protein, fat, and carbohydrates. According to the Chinese recommended reference values of energy-supplying LMs , a reasoThe quality of each included article was evaluated using the Agency for Health Care Research and Quality (AHRQ) scale, which is the cross-sectional study evaluation criteria. The scale contained 11 items; if each item was “yes”, one point would be obtained when each item was “yes”, while “zero” points would be obtained if it was “no” or “unclear”. In total, a score of 8–11 points indicated high quality, 4–7 points indicated moderate quality, and a score of 0–3 points indicated low quality .I2 was used to quantify the degree of heterogeneity, and a high degree of heterogeneity could be confirmed when the I2 value was greater than 75%. The Egger test was used to evaluate publication bias . Revman 5.4 software was used to compare the macronutrient contents of breast milk in high-fat and rational-fat dietary patterns.Protein, fat, and lactose in breast milk were selected as the evaluation indices. The mean differences, 95% confidence intervals (CI), and effect sizes of these macronutrient contents were calculated. Stata 20.0 software was used for meta-analysis, including heterogeneity testing, sensitivity analysis, meta-regression, subgroup analysis, and the evaluation of publication bias. A random-effects model was used to estimate the weighted mean. The statistical results of After the database and article reference lists had been searched, 59 potentially relevant articles were retrieved. Twenty-seven articles met the inclusion criteria, and most of the studies were finally excluded for reasons provided in The results of the mean values of protein content in breast milk at different dietary patterns are shown in The results of the mean values fat contents in breast milk at different dietary patterns are shown in The results of the mean values of lactose contents in breast milk at different dietary patterns are shown in The results of energy supply of macronutrients are shown in I2 values were higher than 75% (p < 0.10), which indicated there was a high heterogeneity. Next, subgroup analyses and meta regression were carried out to explore the source of heterogeneity.Nevertheless, the results showed most I2 values at levels of carbohydrate intake, analytical method, and region.Four variables, including carbohydrate intake, total energy, region, and analytical method, were set as covariates to further identify the sources of heterogeneity. The total energy variable was firstly excluded from further analysis because it could not explain any resources of the heterogeneity. The other results indicated significant differences in I2 value of protein content in the high-carbohydrate subgroup decreased from 99.9% to 49.6%. While it decreased from 98.0% to 78.1%, and to 0.0%, respectively, in the rational-carbohydrate and low-carbohydrate subgroups. In the low-carbohydrate subgroup at the HR pattern, the I2 value of lactose decreased from 97.6% to 81.2%. The results of other subgroups showed no significant difference.As depicted in I2 value of protein content decreased from 99.9% to 85.1% and from 99.6% to 24.2%, respectively. At the HH and RH dietary patterns, the I2 value of fat content decreased from 98.0% to 0.0% and from 99.6% to 0.0%, respectively, and at the HR dietary pattern, the I2 value of lactose decreased from 99.7% to 14.4%. The results of other subgroups showed no significant difference.Based on our results presented in I2 value of protein content decreased from 99.0% to 85.2%, and in the North and South subgroups at the RR pattern, the I2 value of protein content decreased from 74.8% to 0.0%, respectively. In the West and EU subgroups of the HR dietary pattern, the I2 values of protein content decreased from 99.8% to 85.4% and 85.1%, respectively. In the North subgroup of the RH dietary pattern, the I2 values of protein content and fat content decreased from 99.6% to 46.2% and 88.5%, respectively. In the EU subgroup of HH and RR dietary patterns, the I2 value of lactose content decreased from 99.7% to 14.4%, and 99.1% to 14.4%, respectively. These differences might be caused by dietary culture and habits in different regions.Subgroup analysis results showed that the different regions significantly influenced the heterogeneity of breast milk content as described in p < 0.01, respectively).The results of meta-regression, as presented in p > 0.05), except the p value of the protein item at the RH dietary pattern (p < 0.05). Trim and fill analysis also showed that the effect of publication bias was small, indicating that the results above were highly reliable.Egger’s linear regression test showed that there was no significant publication bias (p < 0.05) at the high-fat pattern than that at the rational-fat pattern , but the protein and lactose contents were lower (p < 0.05) at the high-fat pattern than at the rational-fat pattern .The forest map is shown in The purpose of this meta-analysis was to explore the relationship between different dietary patterns and the macronutrient composition of breast milk within the first 1–3 M. Many researchers have investigated the nutrient contents of breast milk in different stages . However, most previous narrative studies focused on a simple dietary influence on selected breast-milk composition, and they did not systematically assess the relationship between diet patterns and breast milk composition based on energy-supplying ratios ,63,64. TThe present study demonstrates that breast milk components, especially fat content, are affected by different dietary patterns. Compared with the other macronutrients, fat is the most variable component of human milk, which is consistent with results of the previous study . The fatOn the other hand, we might judge whether the diet pattern of LMs is rational (RR) or irrational (high protein or high fat) through detecting and comparing macronutrient mean values in breast milk with the estimated mean values since the energy ratios of macronutrients  in breast milk in a reasonable dietary pattern were closest to the estimated energy ratio of macronutrients in infants aged 0–6 M. The results of this study provide new objective evidence to guide the rational diet of LMs, which could further guide nutritional interventions if the LMs’ diet pattern was irrational. The American Heart Association (AHA) also did not recommend the high-protein and/or high fat-pattern since this probably limits the effectiveness of the body’s absorption of essential nutrients . Thus, iHowever, these findings should be interpreted with caution because of their limitations. Firstly, in addition to dietary patterns, many external factors influence the components of breast milk. Therefore, the different contributions of other influencing factors should also be considered though significant heterogeneity exists among the included studies. Although the carbohydrate intake level, region, and analyzer methods can explain parts of the heterogeneity, the residual heterogeneity needs to be explored and evaluated further in the future. Furthermore, because the composition of breast milk is a dynamic process to meet the accelerating growth of infants, the studies we included were cross-sectional studies in nature, analyzing only 1–3 M of mature milk but excluding colostrum and transitional milk. Most researchers have reported that protein content decreases with prolonged lactation, whereas the fat and lactose contents peak and then decrease with prolonged lactation ,14,17,18Additionally, only the relationships between LMs’ diet and macronutrients in breast milk were investigated in this study. In fact, amino acid patterns, fatty acid composition, and different oligosaccharides in breast milk will also have vital impacts on infant development. Therefore, the effect of the LMs’ diet on these elements in breast milk should be considered in future research.This meta-analysis provided a valuable range for breast milk composition at different dietary patterns, and it provided an objective basis for guiding a reasonable diet and avoiding high-protein and/or high-fat patterns during lactation. By detecting the contents of macronutrients in breast milk, we could propose the appropriate dietary intake level of LMs, which may guide the lactation diet scientifically. The results recommend taking a reasonable diet and avoiding a high-fat diet during lactation.


PeKFBs were differentially expressed in various tissues of moso bamboo. All the promoters of the PeKFBs contained stress-related cis-elements, which was supported by the differentially expression of PeKFBs of moso bamboo under drought and cold stresses. Sixteen proteins were screened from the moso bamboo shoots’ cDNA library using PeKFB9 as a bait through a yeast two-hybrid (Y2H) assay. Moreover, PeKFB9 physically interacted with PeSKP1-like-1 and PePRX72-1, which mediated the activity of peroxidase in proteolytic turnover. Taken together, these findings improved our understanding of PeKFBs, especially in response to stresses, and laid a foundation for revealing the molecular mechanism of PeKFB9 in regulating lignin polymerization by degrading peroxidase.The Kelch repeat F-box (KFB) protein is an important E3 ubiquitin ligase that has been demonstrated to perform an important post-translational regulatory role in plants by mediating multiple biological processes. Despite their importance, KFBs have not yet been identified and characterized in bamboo. In this study, 19 PeKFBs were identified with F-box and Kelch domains; genes encoding these PeKFBs were unevenly distributed across 12 chromosomes of moso bamboo. Phylogenetic analysis indicated that the PeKFBs were divided into eight subclades based on similar gene structures and highly conserved motifs. A tissue-specific gene expression analysis showed that the  Post-translational modification (PTM), which plays a central role in the maintenance of protein stability and the modulation of protein activity, affects many aspects of plant development and growth ,2. VarioThe specificity of protein ubiquitination is mainly determined by E3, and due to this, E3 provides recognition and binding specificity to the substrate in a temporally and spatially regulated manner ,6. The EOryza sativa, Arabidopsis thaliana, Triticum aestivum, and Salvia miltiorrhiza respectively [In plants, the KFB subfamily includes members with one to five Kelch repeats consisting of 44–56 residues . To dateectively ,12,13. Sectively , hypocotectively ,16, paniectively , leaf seectively , and othectively ,20. Impoectively . These pectively ,23,24 anectively ,26,27,28ectively , phenolsectively , flavonoectively , and gluectively . Lastly,ectively ,34,35. IPeLAC20” module for lignin monomer polymerization in bamboo was proposed in [To date, it has been challenging to face the depletion of forest resources and the increase in the protection of existing resources, so there is a need to develop and utilize nonwoody biomass . In thisposed in . HoweverThrough BLASTP with the queries of Arabidopsis and rice KFBs in the bamboo database, we looked for conserved F-box domains and Kelch domains by Pfam and SMART in all the candidates and excluded any gene that encoded truncated KFB proteins. As a result, a total of 19 Kelch repeat F-box (KFB) proteins were obtained in moso bamboo (named as PeKFB1-PeKFB19) and used for further analysis. The characteristics of the proteins encoded by PeKFBs, including the numbers of amino acids and Kelch motifs, their molecular weight, and isoelectric point, were systematically analyzed and are listed in To gain clear insight into the evolution of the PeKFBs, a phylogenetic analysis was performed using the protein sequences of KFBs in moso bamboo, Arabidopsis, and rice. The results showed that all the KFBs from different plant species could be clearly demarcated into eight major subclades and that the members of each subclade were different . Within PeKFBs had no introns while the rest of the PeKFBs only contained one intron , followed by chromosome 17 with three PeKFBs and chromosome 5 with two PeKFBs, while the other chromosomes anchored only one. To understand the duplication events of KFBs in moso bamboo and rice, a synteny analysis was performed using MCscanX and the Advanced Circos software in TBtools. The results demonstrated that nine segmental duplication pairs were found within PeKFBs and 23 segmental duplication pairs were detected between PeKFBs and OsKFBs  in the heat map according to the hierarchical clustering of expression patterns; the genes in Group I–Group III generally showed a higher expression abundance than those in other groups. Group I consisted of four PeKFBs that had a higher expression level in young tissues. For example, PeKFB9 showed a high expression in 0.2 m shoots, 0.1 cm roots, and buds; however, its expression pattern was not significant in Group II. Group III included five PeKFBs (PeKFB2/10/14/16/17) that had high transcript levels in all stages and tissues. A relatively low transcript expression level of these genes was observed in older roots, shoots, and bud tissues; a high expression abundance of them was detected in 0.2 m shoots and rhizome. Similarly, they showed higher expression in the top and middle than those in the low parts of single internodes (PeKFBs in Groups I-III might be directly or indirectly involved in the rapid growth of bamboo shoots. Group IV included only two members (PeKFB6 and PeKFB7) that were almost undetected in all tissues. The PeKFBs in Group V showed obvious tissue-specific effects correlated with the different developmental stages and the tissues. For instance, PeKFB13 showed a relatively dominant expression level in blades but a relatively lower one in the other tissues.The gene expression patterns could provide information for potential gene functions. The recent transcriptome data generated from different tissue samples was used to investigate the expression patterns of the PeKFBs . Accorditernodes . These rPeKFBs without suitable specific primers, 15 PeKFBs were carefully selected for RT-qPCR analysis according to their specific expression patterns in different tissues  . The dedIn addition, another candidate protein (PH02Gene23181) was identified by the Y2H screening that belonged to the peroxidase protein family and was homologous with AtPrx72, which had been shown to participate in lignin polymerization , so we nFurthermore, we verified whether the interaction of PeKFB9 with the PePRX72-1 decreased peroxidase activity in order to evaluate the hypothesis that PeKFB9 mediated the degradation of the peroxidase. The assay of the transient expression of PePRX72-1 in tobacco leaves showed that co-expression of the PePRX72-1-FLAG fusion proteins with PeKFB9 decreased peroxidase activity more than 60% compared with the control a. To verKFB proteins play crucial roles in the course of plant growth and in the response to biotic and abiotic stresses, as well as in phenylalanine biosynthesis ,45. LignArabidopsis, rice, potato, wheat, and chickpea [We identified 19 PeKFBs in the genome of moso bamboo, which was less than the numbers previously found in chickpea ,45,47,48chickpea . In addichickpea . In contchickpea , KFBs inPeKFBs in different tissues and found a large variety among different PeKFBs  . Therefore, it was consistent with the results of previous studies that these genes may be involved in the abiotic stress response and participate in the regulation of ABA signaling [cis-elements were distributed in the promoters of a few PeKFBs, which was consistent with the results showing that the transcriptions of the PeKFBs were not responsive to these phytohormones  had the same structure but showed different expression patterns  , suggestin genes ,51. Amon signals ,53. ABREignaling ,55. Meanhormones , but didpatterns , which wArabidopsis, which altered cell wall and phenylpropanoid metabolism. Class III peroxidases are glycoproteins with a major role in lignin formation and have been implicated in the cross-linking of lignin formation [Arabidopsis were implicated in lignification, including AtPRX2, AtPRX4, AtPRX17, AtPRX25, AtPRX52, and AtPRX71, due to altered lignin content or composition in mutant plant lines [Lignification is regulated in transcriptional regulation, translational regulation, post-translational modification and proteolysis, and product inhibition. With regard to lignification in bamboo, most studies focused on transcriptional regulation ,42. To dormation ,57. In rormation . Severalnt lines ,60,61,62https://pfam.xfam.org, accessed on 19 April 2022, European Molecular Biology Laboratory, Cambridgeshire, UK) to detect the presence and number of the F-box and Kelch domains. All KFB sequences were manually inspected and those that did not contain the conserved domains were discarded. Finally, the KFB proteins were named according to their BambooGDB assembly names in order.To obtain nonredundant KFB proteins in moso bamboo comprehensively, a BLASTP algorithm-based search provided by the BambooGDB database under the E-value cutoff of 1e-10 was first performed using the KFB protein sequences reported in Arabidopsis and rice . The reshttp://www.csbio.sjtu.edu.cn/bioinf/plant-multi/# accessed on 19 April 2022).The basic characteristics of the identified KFB proteins in moso bamboo were further analyzed, including the predicted proteins and the physicochemical parameters. The predicted molecular weights (MWs) and isoelectric points (pIs) of the KFB proteins were analyzed using ProtParam . The subcellular localization was predicted by using Plant-mPLoc (Arabidopsis genome from TAIR (The Arabidopsis Information Resource) release 10.0 (http://www.arabidopsis.org accessed on 19 April 2022) and the rice genome annotation database  respectively, which were used for the construction of the phylogenetic trees.To explore the evolutionary relationships among the KFBs in moso bamboo, MEGA 6.0 was used to perform multiple sequence alignments with the full length of KFB amino acid sequences and the maximum-likelihood phylogenetic unrooted trees were constructed using the FastTree program . The KFBhttp://meme-suite.org accessed on 19 April 2022, National Institutes of Health, Bethesda, Rockville, MD, USA) was adopted to identify the conserved motifs with the following parameters: the distribution of motif occurrences—zero or one per sequence; the maximum number of motifs—10; and the optimum width of each motif—between 6 and 300 residues [The gene structure of the KFB genes in moso bamboo was predicted using TBtools software . The GFFresidues .Ka) and synonymous substitution rates (Ks), as well as the Ka/Ks ratios of syntenic KFB gene pairs, were calculated using KaKs_Calculator software [The chromosomal localization data of each KFB gene identified in moso bamboo was retrieved from the GFF file using TBtools. MCScanX software was used to identify the duplicated and syntenic KFB genes within the moso bamboo genome and those between moso bamboo and rice using the default settings . The chrsoftware .http://bioinformatics.psb.ugent.be/webtools/plantcare/html/ accessed on 19 April 2022). The number of elements in the promoters was shown in heat map made by using TBtools software.To obtain the promoter sequences of KFB genes in moso bamboo, we downloaded 2000 bp of the sequence upstream of each KFB gene coding region from the BamGDB. Cis-elements were analyzed using the database of PlantCARE  and loaded into TBtools software to generate heat maps. To investigate the expression of KFB genes in the different tissues and those in shoots at different developmental stages of moso bamboo, the RNA-seq data generated from 26 samples of rhizomes, roots, shoots, leaves, sheaths, and buds were downloaded from the NCBI Short Read Archive (SRA) and used for further analysis. The accession numbers in SRA were SRX2408703~SRX2408728 and SRR13201212-SRR13201244, respectively. The RNA-seq data of moso bamboo leaves under NAA and GASample collection: moso bamboo seeds gathered from the Guangxi Province of China were sown and cultivated in a greenhouse . After three months, the seedlings were used for the following treatments. Drought and cold stresses were simulated using a 20% PEG 6000 solution and 4 °C, respectively. The solution was poured into the cultured pots of moso bamboo seedlings and the leaves were harvested at 3 h, 6 h, and 12 h, respectively, and immediately frozen in liquid nitrogen and stored at −80 °C for RNA isolation. Meanwhile, the untreated seedlings were also collected and used as the control (0 h).RNA extraction: the total RNA was extracted using a plant RNA extraction kit  according to the manufacturer’s instructions. The integrity of the total RNA was verified via agarose gel electrophoresis and the purity and concentration of the total RNA was determined via spectrophotometry . The first strand cDNA was synthesized using a reverse-transcription system . For each 20 μL reaction, 1000 ng of total RNA was used; the synthesis was performed at 42 °C for 45 min. The final cDNA product was diluted 5-fold prior to use.2O. NTB was used as a reference gene [−ΔΔCt method was used for the analysis and visualization of the generated PCR data [RT-qPCR: the specific primers for different KFB genes of moso bamboo were designed in Primer Premier 5.0 software and empirically adjusted for gene expression analysis . Additionce gene  and the PCR data .PeKFB9 (PH02Gene23593.t1) was cloned into the pGBKT7 bait vector  using the cDNA synthesized from the mRNAs of shoots at different developmental stages and cloned into the prey vector pGADT7. The full-length CDS of t vector  and the PeKFB9 and PePRX72-1, as well as the trunked PeKFB9 (F^PeKFB9 and K^PeKFB9), were cloned via PCR using the primers described in ® cloning method to generate PeKFB9-FLAG, F^PeKFB9-FLAG, and PePRX72-1-FLAG, respectively. The Agrobacterium strain GV3101 carrying the PePRX72-1 expression vector (OD600 = 0.6) was mixed with an equal volume of an Agrobacterium strain (OD600 = 0.6) carrying PeKFB9-FLAG, F^PeKFB9-FLAG, and empty pCAMBIA1300-3×FLAG, respectively; these were then used to infiltrate the tobacco leaves.The ORFs of −1. The concentration of total protein was measured by using a Bradford protein assay kit . The proteins were examined using immunoblotting with an anti-FLAG monoclonal antibody.After three days of infiltration, the total soluble proteins were extracted with lysis buffer containing 20 mM of Tris-HCl (pH = 8.0), 150 mM of NaCl, 1 mM of EDTA, 10% glycerol, 0.2% Triton X-100, and 1 × protease inhibitor cocktail. The ratio of leaf fresh weight to extraction buffer was 1 g mlIn addition, harvested leaves (0.1 g) frozen at −80 °C were crushed with a prechilled mortar and pestle using an appropriate volume of prechilled extraction buffer. After uniform homogenization and centrifugation at 13,000 rpm for 15 min at 4 °C, the supernatant containing the crude enzymes was collected and used for the peroxidase activity analyses using the kit and following the manufacturer’s instructions .p < 0.01.Analyses were performed with SPSS Statistics for Windows . All data were the average and standard deviation (SD) of three biological replicates. One-way analysis of variance was used to evaluate the statistical significance of differences among means using SPSS software. Different letters indicate respective significant differences at the level of PeKFBs were mapped to 12 chromosomes unevenly; their exon-intron organization and the conserved motifs in their coding proteins were relatively similar to those in other monocot plants. The expression patterns of the PeKFBs in different tissues and organs at various developmental stages implied that they may be involved in the growth and development of moso bamboo. The cis-elements in the promoter regions of the PeKFBs and their expression patterns indicated that the PeKFBs might be involved in responses to drought and cold stresses. The results of the peroxidase activity and Western blotting analyses indicated that PeKFB9 was a post-translational regulator responsible for the turnover of peroxidase to negatively control the monomer polymerization. Taken together, the results of the present study provided an extensive resource for understanding the diversity of KFB genes that can be utilized for genetic manipulation in improvement of abiotic stress tolerance and wood properties in bamboo.In the present study, we conducted a comprehensive analysis of the KFB family in moso bamboo for first time. A total of 19 PeKFBs were identified and classified into eight subclades. The


Background: Chikungunya is a viral disease that is transmitted by mosquitoes. It is characterized by an acute onset of fever and severe arthralgia. Methods: We describe six cases of acute and post-acute chikungunya in which viral RNA was detected in semen. Conclusions: The most prolonged detection period was 56 days after illness onset. We attempted to cultivate positive semen samples, but virus isolation was unsuccessful in all cases. Alphavirus). CHIKV is transmitted by Aedes aegypti and Aedes albopictus mosquitoes [Chikungunya is a neglected tropical disease caused by chikungunya virus (CHIKV), an RNA arbovirus belonging to the Togaviridae family  RNA in semen have been described in some studies ,7. In adThe knowledge of viral persistence in genital fluids is imperative to elucidate different forms of infection and possible reactional activity in response to the continuity of the virus in the body, which has relevance for the diagnosis and pathophysiology of viral diseases. Confirmation of the detection and persistence of the virus in semen is useful in health programs, especially for viruses with high rates of mortality or morbidity .The present study aimed to describe the detection and duration of CHIKV RNA detected in semen samples obtained from six symptomatic men.From 16 April 2019 to 10 October 2019, six male patients infected by CHIKV, and with the virus detectable in semen samples using real-time reverse transcriptase polymerase chain reaction (rRT-PCR), were followed for 3 months at the Acute Febrile Illnesses outpatient clinic of Oswaldo Cruz Foundation in Rio de Janeiro, Brazil. The patients were enrolled in a cohort study to evaluate the presence and duration of CHIKV infection in bodily fluid samples obtained from adult patients. The local ethics committee reviewed and approved the study (CAAE:06779019.0.0000.5262). Clinical data and samples  were collected at enrollment and every 15 days for 2 months, with a final collection at 3 months, for a total of six visits .The samples were tested for CHIKV, ZIKV, and dengue virus (DENV) using rRT-PCR. RNA was extracted using the QIAmp Viral RNA Mini Kit. The general procedures for rRT-PCR for chikungunya, Zika, and dengue have been described elsewhere ,15,16. T2 flasks under four conditions: 1. The diluted inoculum was filtered with 0.22 µm syringe filter and removed from washing with 199 medium without FBS after adsorption; 2. The diluted inoculum was filtered with a 0.22 µm syringe filter and maintained after adsorption; 3. The diluted unfiltered inoculum was removed by washing with 199 medium without FBS after adsorption; 4. The diluted unfiltered inoculum was maintained after adsorption. All inoculums were adsorbed to the cell monolayer for one hour at 37 °C/5% CO2. To establish the conditions for viral isolation in cell culture, three semen samples with Ct 32, and one with Ct 34, with sufficient sample volume for testing and subsequent extractions, were initially diluted (1:4) in culture medium 199 without fetal bovine serum (FBS). Then, 100 µL was inoculated in a monolayer of VERO cells  and grown in 12.5 cm2 for up to 14 days and monitored daily for cytopathic effects using an inverted microscope. Aliquots of 140 μL of supernatants from all flasks were taken at 3, 7, and 14 days of incubation, and were subjected to viral RNA extraction using the QIAamp Viral RNA Mini Kit , according to the protocol described by the manufacturer, and stored at −70 °C. RNAs was tested for the detection of CHIKV by rRT-PCR, as described previously [After incubation, 1.5 mL of culture medium 199 containing 2.5% FBS was added. Flasks containing only the cell monolayer without manipulating the medium exchange were used as negative controls during the inoculation process. All flasks were incubated at 37 °C/5% CO2 flasks. The inoculums were not filtered and were maintained after adsorption. All inoculums were adsorbed to the cell monolayer for one hour at 37 °C/5% CO2, and, after incubation, 1.5 mL of culture medium 199 containing 2.5% FBS was added. Flasks were incubated at 37 °C/5% CO2 for up to 14 days and monitored daily for cytopathic effects using an inverted microscope. Aliquots of 140 μL of supernatant from all flasks were taken at 3, 7, and 14 days of incubation and were subjected to viral RNA extraction using the QIAamp Viral RNA Mini Kit , according to the protocol described by the manufacturer, and stored at −70 °C. RNA was tested for the detection of CHIKV by rRT-PCR, as described previously [After testing the viral isolation conditions, all semen samples that were previously positive by rRT-PCR were inoculated in cell culture. The eight samples were diluted in culture medium 199 without FBS, six at a ratio of 1:2 (50 μL of semen + 50 μL of culture medium), and two samples at 1:10 (10 μL of semen + 90 μL of culture medium), owing to the scarcity of the original sample. A volume of 100 μL was inoculated into a monolayer of VERO cells grown in 12.5 cmAll patients presented with symptomatic disease and moderate clinical manifestations. Their median age was 43.5 years . There were no significant changes in the hematological or biochemical parameters at any of the study visits. All patients had chikungunya confirmed by detectable rRT-PCR in the serum during enrollment or seroconversion of anti-CHIKV-IgM. Patient 1: A 56-year-old male with hypertension and asthma complained of high fever (>40 °C), myalgia, asthenia, taste alteration, and moderate polyarthralgia for 4 days. He also described episodes of vomiting and nausea for 2 days. He developed a cutaneous rash 5 days after symptom onset. After the acute phase, mild symmetrical arthralgia in the shoulders, knees, and ankles resolved within six weeks without anti-inflammatory drug use.Patient 2: A 48-year-old male previously healthy, with a 3-day history of fever, chills, headache, severe arthralgia, and prostration. On the seventh day, he developed a diffuse rash on the trunk, abdomen, and limbs. Joint pain was symmetrical  and lasted for more than six weeks. The patient was treated with prednisone (20 mg/day for 7 days) with a favorable outcome. Coinfection with DENV type 2 was detected.Patient 3: A 51-year-old male with hypertension presented with a 4-day history of acute fever, malaise, generalized rash, and pain in multiple joints. The patient presented with edema in the hands and knees. The polyarthralgia was severe, lasted throughout the study period (three months), and was localized mainly in the knees. He was administered prednisone (20 mg/day for 14 days) and gabapentin (300 mg/day for 30 days) with partial improvement of his arthralgia.Patient 4: A 33-year-old previously healthy man developed an abrupt onset of fever, chills, prostration, and moderate polyarthralgia affecting the shoulders, wrists, proximal interphalangeal joints, knees, ankles, and metatarsophalangeal joints. He had a 3-day history of a disseminated maculopapular rash. The fever lasted for 3 days, and mild joint pain lasted 8 weeks, without any pharmacological intervention. Patient 5: A 39-year-old man with three days of fever, chills, and sweating associated with severe pain in the shoulders, wrists, knees, ankles, and feet. Exanthema presented for 3 days. Polyarthralgia was severe and persisted throughout the follow-up period. Prednisone (20 mg/day for 20 days) was prescribed for partial pain relief. Patient 6: A 35-year-old man with a 5-day history of moderate fever, headache, prostration, myalgia, joint pain, and exanthema. The polyarthralgia was severe and lasted for more than 8 weeks. The patient received prednisone (20 mg/day for 21 days) with partial clinical relief of his symptoms. Twenty-three semen samples were collected from these six patients, and eight were CHIKV-rRT-PCR-positive . The maxVirus IsolationFrom the initial experiment to test the best conditions for viral inoculation, it was possible to recognize that there was no need for the filtering step of the semen samples to minimize possible bacterial contamination, since the cell cultures did not present typical turbidity, maintaining the transparent and translucent supernatant throughout the incubation period. After 3 days of inoculation, one of the four samples showed the beginning of morphological changes in the cell monolayer, resembling the expected cytopathic effect, progressing gradually over the next few days, with cell detachment after 7 days of inoculation. Such evolution initially occurred in cultures with diluted inoculums with and without filtration, and was maintained after adsorption. The same was observed later in the flasks that received the diluted inoculum of the same sample, with and without filtration, which was washed after adsorption. However, the rRT-PCR results of the supernatants from all samples collected at three points after incubation were negative. rRT-PCR of the original sample corresponding to the inoculations that would seem to progress in cytopathic effect was performed, but the result was negative.A new viral inoculation was carried out with eight semen samples available under the conditions described in the methodology section. Three days after inoculation, two flasks showed complete detachment of the cell monolayer, one of which was inoculated with the sample diluted 1:10. Other cell monolayers began to show morphological changes, with cell detachment beginning 7 days after inoculation, and total detachment after 14 days. rRT-PCR of supernatants collected throughout the incubation period showed amplification with Ct 35 at the three collection points from a single sample initially diagnosed with Ct 28, which was considered as negative isolation, since the highest Ct maintained throughout the incubation period possibly refers to the inoculum remnant. All supernatants from the other samples tested negative.In this case series, CHIKV could be detected in semen samples from patients with either acute or convalescent disease. The longest CHIKV RNA detection period was 56 days after symptom onset in a 33-year-old, previously healthy man. All virus isolation attempts were unsuccessful for all samples.Efforts to demonstrate virus detection in genital secretions of individuals infected with viruses that had previously been unknown to be sexually transmitted are necessary to investigate additional forms of transmission that could be of public health importance. Sexual transmission of Ebola virus was confirmed in Liberia in 2015 . Sexual Bandeira et al.  describeAlthough CHIKV RNA was detected in the semen in our case series, the virus was not detected by rRT-PCR in the cultured samples. One explanation is that the cycle threshold values found in our samples were high, ranging from 28 to 34, which may correspond to a low viral load in semen. Viral cultures perform better with low Ct values in inoculated samples . For exaFactors intrinsic to the nature of the biological sample can influence the degradation of nucleic acid viruses by the presence of nucleases and the integrity of the viral particle, which can suppress viral infection and produce cytotoxic substances for cell culture ,25. In aAdditional limitations of our study were the small sample size and the short follow-up period. Infectivity is a prerequisite for pathogen transmission, and depends on factors such as the infectious dose and exposure route. Therefore, virus isolation remains the only direct and definitive approach for proving infectivity . Twenty-Studies on viral detection and semen persistence benefit clinical practice and public health, especially for viruses that can cause high chronic morbidity, such as CHIKV. Further studies are needed to evaluate the potential infectivity of semen and the sexual transmission of chikungunya.


Animal breeders are always interested in improving the economically important traits such as growth by selecting and breeding genetically superior animals. The Lohi sheep is one of Pakistan’s most important meat type breeds as it significantly contributes to the country’s total mutton production. To fulfil the higher demands of mutton production, there is a need to define optimum breeding strategies for improving growth in this breed. This study reported the moderate heritabilities for growth traits in Lohi sheep and significant influences of sex, year of birth and type of birth on these traits. The results might be useful for designing a breeding program for higher mutton production in Lohi sheep.Estimating genetic parameters for growth traits is crucial to plan breeding strategies for improving meat production in indigenous sheep breeds. The study first tested the effects of environmental and maternal effects on five growth traits, including birth weight (BWT), weight at 120 days (WT120), weight at 180 days (WT180), weight at 270 days (WT270) and weight at 365 days of age (WT365) and then estimated genetic parameters for these traits using data obtained in 1215 Lohi sheep. The effects of factors, including year (YOB), season (SOB) and type of birth (TOB), age of dam (AOD) and sex on growth traits of Lohi sheep, were examined using analysis of variance (ANOVA) in R software. Sex, TOB and YOB significantly affected all studied traits. The estimates of direct and maternal heritability for BWT and WT120 were 0.15 ± 0.08 and 0.20 ± 0.06, and 0.45 ± 0.16, 0.21 ± 0.08, respectively. The direct heritability estimates for WT180, WT270 and WT365 were 0.20 ± 0.07, 0.21 ± 0.07 and 0.19 ± 0.08, respectively. Due to the high heritability estimate obtained for WT120 compared to other studied traits, and its strong genetic correlation (>0.9) with post-weaning growth traits, it is recommended that selection must be practiced on WT120 to improve the growth performance of Lohi sheep. The results could be used for the development of genetic/genomic selection programs aiming to improve the production performance of the Lohi sheep. The production of animals with higher growth rates is necessary to fulfil the increasing need for animal protein in the human diet. Growth of an animal is defined as the increase in size or volume of tissues over time ,2. The gThe Lohi, a well-known thin-tailed sheep breed of Pakistan, is known for its mutton and wool production. It can be found throughout the Punjab province, but most of its population is in central Punjab. It contributes up to 40% of the total mutton production of the province ,17. It sThe study was executed at the Small Ruminant Training and Research Centre (SRT & RC) Pattoki, Punjab, Pakistan. Lohi sheep farming at SRT & RC was started in 2007 by purchasing 100 animals from Livestock Experiment Station, Bahadurnagar, Okara. The average age of Lohi lambs at weaning was four months. After weaning, males and females were separately raised in different sheds. Controlled breeding was practised on the farm. Males were exposed daily to females for 1 h during breeding seasons: Spring (February to April) and Autumn (September to November). The females that did not conceive during breeding seasons were again exposed to males. Animals were allowed to graze for 4 to 5 h daily on shrubs and seasonal forages. Seasonal forages were also provided in the chopped form to the animals after they returned to grazing. Additionally, concentrates were also provided to the pregnant, suckler and breeding males at the rate of 250 to 500 g/day, 100 to 150 g/day and 800 to 1000 g/day, respectively.The data regarding animal ID, birth type, sex, date of birth, pedigree and body weights on 1215 Lohi lambs born from 2008 to 2019 were collected from SRT & RC. The data were evaluated for the identification of outliers. The growth records which lied outside the mean ± 3 standard deviations were removed before the final analysis.Because of variation in weaning ages, body weights were adjusted to 120 days of age. Similarly, due to age differences in days at 6, 9 and 12 months, body weights were corrected to 180, 270 and 365 days of age using the formula given below:z = adjusted weight for specific days of age ,b = birth weight,w = weight recorded at 4, 6, 9 and 12 months,a = age in days at the measurement of weight at 4, 6, 9 and 12 months,d = 120, 180, 270 and 365 days accordingly for each trait.p < 0.05) were used in models for the estimation of variance components. The least square (LS) means for each category within each factor were estimated using the lsmeans package of R [The impact of different environmental sources of variation on growth traits, including birth weight (BWT), weaning weight (WT120), weight at 180 (WT180), weight at 270 (WT270) and weight at 365 days of age (WT365) were investigated through the analysis of variance (ANOVA) using the ‘car’ package  in R verage of R . Duncan’age of R .We used six animal models accounting for or ignoring maternal permanent environmental effect, maternal genetic effect or their combination in the univariate analysis, performed through the restricted maximum likelihood method (REML) of WOMBAT to estimate covariance components and heritability of growth traits . Model 1Model 2Model 3Model 4Model 5a, m is the vector of maternal additive genetic effect and Zm is the incidence matrix linked to it, c is the vector of maternal permanent environmental effect with incidence matrix Zc, e is the vector for residual effects, A is the numerator relationship matrix between animals and Model 62) was utilised to announce the significant variation between the two models and was calculated using the formula given below as described by [The log-likelihood ratio test was applied to find an appropriate animal model for each growth trait. The Chi-square test  variation in the body weights of lambs born in different years could be explained due to differences in environmental conditions, management of the farm and the availability of forages throughout these years. Factors like climatic conditions, availability of nutrients, fodder quality and disease prevalence can affect an animal’s growth performance. The maximum performance of an animal can only be attained if it is provided with good-quality nutrients, comfortable surroundings and disease control. As farm resources did not remain constant during the study period (12-year span), the availability of resources could have influenced the decisions of the farm manager, and, as a result, animal management was affected. Similarly, the availability of grasses to the lambs for grazing also depends on various factors, including rainfall. Hence, collectively all of these factors could have contributed to the significant variation in lambs’ growth performance in different years of study. The significant variation in growth traits of lambs born during different years observed in the current study was similar to previous studies on sheep [The descriptive statistics of growth traits of Lohi sheep are shown in on sheep ,27,28,29Trifolium alexandrinum) and oats (Avena sativa) were excessively available during the winter and spring seasons at the farm. Hence, dams which completed their gestation during these seasons had a better chance to avail more fresh fodder, which had an effect on their milk performance and lambs belonging to these dams were found heavier at weaning. The significant influence of SOB on some growth traits agreed with previous findings [The studied growth traits, except BWT and WT365, were significantly affected by SOB . The insfindings ,33,34,35Lamb’s sex significantly influenced all the studied traits . Male laDam’s age only significantly affected weights at early ages, i.e., BWT, WT120 and WT180 . Some otWe observed a significant influence of TOB on BWT, WT120, WT180, WT270 and WT365 . The sinThe log-likelihood test revealed Model 3 as the best equation for BWT and WT120 . This moThe estimate of direct heritability attained by model 3 for BWT was 0.15 ± 0.08. These results were close to values of 0.10 and 0.11 reported by Babar et al.  and JaveThe direct and maternal heritability estimates for WT120 were high and moderate, respectively. The high heritability for WT120 indicates that selection for heavy-weight animals at weaning will improve the growth performance of Lohi sheep. Similar to the present study, higher direct heritability estimates were observed in Madras Red, Menz and Djallonke sheep ,50,54. CThe magnitude of the relationship between direct and maternal genetic effects for BWT and WT120 was highly negative, indicating the significant role of maternal genetic effect in the pre-weaning growth traits of Lohi sheep. It also suggests that improving one effect consequently decreases the other one. According to Szabó et al. , by selePost-weaning growth traits of Lohi sheep were not significantly affected by maternal genetic and permanent environmental effects, limiting maternal effects to pre-weaning stages. The direct heritability for WT180 was estimated as 0.20 ± 0.07, similar to the findings of Kumar et al.  in DeccaThe heritability estimate (0.21 ± 0.07) for WT270 in the current study was in the range of earlier reports. The estimates for Makouei, Malpura, Muzaffarnagari, Zandi and Nellore sheep ranged between 0.1 to 0.16 ,41,53,61The estimate of heritability for WT365 was 0.19 ± 0.08, which was in the range from 0.14 to 0.43 reported in other sheep breeds, including Muzaffarnagari, Ghezel, Djallonke, Pak-Awassi, Nilagiri, Harnali, Deccani, Horro and Harnai ,66,69,71Several bivariate analyses were performed to estimate the genetic and phenotypic correlations between the growth traits of Lohi sheep. The results of the analyses are represented in The results suggested that environmental factors like SOB, YOB, AOD, TOB and sex were significant sources of differences in the growth performance of Lohi sheep. So, it is recommended that known environmental factors must be considered while defining a model to estimate genetic parameters. The maternal effect only influenced the pre-weaning growth traits and did not influence post-weaning growth traits, suggesting that the effect of maternal components should be considered to estimate unbiased genetic parameters for pre-weaning growth traits. The highest heritability estimate obtained for weaning weight and strong genetic correlation of weaning weight with post-weaning traits suggests that selection could be fruitful if it is based on the weaning weight of Lohi sheep.


Non-small cell lung cancer (NSCLC) is the most common type of lung cancer, accounting for approximately 80%-85% of all lung cancer cases. This study aimed to investigate the value of diffusion kurtosis imaging (DKI), diffusion-weighted imaging (DWI) and 2-[18F-FDG PET/MR. Mean kurtosis (MK), mean diffusivity (MD), apparent diffusion coefficient (ADC), maximum standard uptake value (SUVmax), metabolic tumor volume (MTV) and total lesion glycolysis (TLG) were measured. The independent samples t test or Mann–Whitney U test was used to compare and analyze the differences in each parameter of SCC and AC. The diagnostic efficacy was evaluated by receiver operating characteristic (ROC) curve analysis and compared with the DeLong test. A logistic regression analysis was used for the evaluation of independent predictors. Bootstrapping (1000 samples) was performed to establish a control model, and calibration curves and ROC curves were used to validate its performance. Pearson’s correlation coefficient and Spearman’s correlation coefficient were calculated for correlation analysis.Seventy-seven patients with lung lesions were prospectively scanned by hybrid 3.0-T chest The MK and ADC values of the AC group were significantly higher than those of the SCC group , and the SUVmax, MTV, and TLG values of the SCC group were significantly higher than those of the AC group . There was no significant difference in the MD value between the two groups. Moreover, MK, SUVmax, TLG and MTV were independent predictors of the NSCLC subtype, and the combination of these parameters had an optimal diagnostic efficacy , which was significantly better than that of MK , ADC , SUVmax , MTV  or TLG . The ROC curve showed that the validation model had high accuracy in identifying AC and SCC ;. The SUVmax value was weakly positively correlated with the Ki-67 index , the ADC and MD values were weakly negatively correlated with the Ki-67 index , and the MTV and TLG values were weakly positively correlated with NSCLC stage .18F-FDG PET are all effective methods for assessing the NSCLC subtype, and some parameters are correlated with stage and proliferation status.DKI, DWI and  Ultimately, a total of 77 patients were included in this study. There were 48 males and 29 females, with an age of 62.73 ± 9.06  years, (This study was approved by the Research Ethics Committee (RCE) of our hospital. After receiving written informed consent from each patient, prospectively surveyed patients who were suspected of having a space-occupying lesion of the lung through CT examination in our hospital from July 6, 2020, to June 29, 2021, were enrolled. A total of 120 patients underwent chest ) years, Figure 118F-FDG was produced by a cyclic accelerator and an automatic chemical synthesis system, and the radiochemical purity of the tracer was >95%. It was recommended that patients should not exercise vigorously within 24 hours before the examination and should fast for more than 6 hours to achieve a fasting blood glucose in the morning of< 7 mmol/L. The tracer was injected intravenously at the standard dose of 0.11 mCi per kilogram of body weight, and images were collected at a resting time of approximately 60 minutes  with a 12-channel phased array body coil. The PET tracer  minutes  after inThe collected PET/MR images were transferred to the United Imaging Workstation , and the workstation automatically performed image registration. For PET images, the PET/MR fusion software was used to post-process the metabolic parameters and automatically extract the volume of interest (VOI) of the tumor. Using 40% SUVmax as the threshold, the maximum standard uptake value (SUVmax), metabolic tumor volume (MTV) and total lesion glycolysis (TLG) of the tumor were automatically calculated. For DWI and DKI images, the parameters were processed using the workstation’s advanced analysis toolkit. The DWI parameter was generated using the following formulae:0 is the signal strength when the b value is 0 s/m2, Sb is the signal strength at different b values, b is the diffusion sensitivity value, and ADC is the apparent diffusion coefficient. The DKI parameters were generated using the following formulae:SKapp represents the apparent kurtosis coefficient, Dapp represents the corrected apparent diffusion coefficient, and MD and MK are the mean Dapp and Kapp values of all directions. Using the PET/MRI fusion image as a reference, we manually delineated the ROIs (regions of interests) on the image of each layer containing the tumor on the axial T2WI image while paying attention to avoid places that are prone to artifacts, such as hemorrhage, necrosis and cystic degeneration. The software automatically copied the delineated ROIs to all parameter maps to calculate the average values based on the gross tumor volume (GTV). All the above quantitative parameters were independently processed and measured by two radiologists  without any clinical or pathological information using a double-blind method.All patients in the study provided pathological specimens, which were obtained by focal biopsy or surgical procedures within 4.19 ± 2.06, (range 1-7) days after the PET/MR scan. The tumor tissue was isolated and sent to the pathology department of our hospital to determine the NSCLC subtype. Formalin fixation, dehydration, wax immersion, embedding, sectioning and routine HE staining were performed. The eighth edition of International Association for the Study of Lung Cancer (IASLC) was used for staging classification . The expSPSS 23.0 software and MedCalc 15.0 software were used for data analysis. The intraclass correlation coefficient (ICC) was used to assess the consistency of each parameter by two radiologists, as follows: r ≥ 0.75, excellent agreement; 0.60 ≤ r< 0.75, good agreement; 0.40 ≤ r< 0.60, fair agreement; and r< 0.40, poor agreement . The ShaThe clinical and pathological characteristics of all the patients are shown in The MK, MD, ADC, SUVmax, MTV and TLG values measured by the two observers all had high consistency, and the ICCs were 0.826, 0.799, 0.819, 0.823, 0.869 and 0.757, respectively. The average of the two observers’ parameter values was taken as the final result and included in this study.18F-FDG PET parameters. The MK and ADC values in the AC group were significantly higher than those in the SCC group , while the values of SUVmax, MTV and TLG in the SCC group were significantly higher than those in the AC group . There was no significant difference in the MD value between the AC group and SCC group , . Multivariate analysis showed that MK, SUVmax, TLG and MTV were independent predictors of NSCLC subtype  Table 4.-3 mm2/s, SUVmax: 7.27 g/cm3, TLG: 19.11 g and MTV: 5.05 cm3.The AUCs of MK, SUVmax, TLG, MTV and ADC used to diagnose SCC and AC were 0.758, 0.740, 0.716, 0.715 and 0.679, respectively , but there was no statistically significant difference in the diagnostic efficacy among these parameters . The cutoff values for each parameter were as follows: MK: 0.43, ADC: 1.270 ×10Independent predictors MK, SUVmax, TLG and MTV were used for combined diagnosis, and the AUC (MK + SUVmax + TLG + MTV) was 0.876 . Moreover, the difference between AUC (MK + SUVmax + TLG + MTV) and AUC (MK), AUC (ADC), AUC (SUVmax), AUC (MTV), AUC (TLG) was significant  Table 5.Bootstrapping with 1000 samples was used to validate the multivariate regression model. The ROC showed that the validation model had high accuracy in identifying AC and SCC . ADC and MD values were weakly negatively correlated with the Ki-67 index . There was no significant correlation between MTV, TLG, MK and the Ki-67 index . The differences between r1 and r2, r2 and r3, and r1 and r3 were not significant  Table 6.There was a weak positive correlation between the MTV and TLG values and NSCLC stage , and there was no significant difference between r4 and r5 . There was no significant correlation between the SUVmax, ADC, MK, and MD values and NSCLC stage  Table 6.ADC and MD all reflect the restricted diffusion of water molecules in the tissue and are closely related to the cell density, intracellular matrix and number of organelles . In thisMK is the most representative parameter of DKI , and its18F-FDG is, the greater the SUVmax value , the National Natural Science Foundation of China (81720108021 and 31470047), the Zhengzhou Collaborative Innovation Major Project (20XTZX05015), Zhongyuan Thousand Talents Plan Project - Basic Research Leader Talent (ZYQR201810117), the Key Project of Henan Province Medical Science and Technology Project (LHGJ20210001 and LHGJ20210005) and Henan provincial science and technology research projects (212102310689).We acknowledge financial support by the National Natural Science Foundation of China.Authors JY, YY, and ZW were employed by United Imaging Healthcare (UIH).The remaining 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. 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