The concerns of food safety are rising in developing countries such as South Africa as a result of heavy metal contamination of culinary herbs and spices. Spices and herbs are used for therapeutic purposes as well as flavoring and coloring food. Heavy metals in spices represent significant health risks due to their high toxicity in high quantities. A total of 20 spices samples were purchased from different registered shops, for heavy metals analysis. The samples were prepared, digested, and analyzed with an inductively coupled plasma mass spectrometer (ICP-MS). To ensure the method's accuracy, Polish Certified Reference Mixed Polish Herbs (INCT-MPH-2) from the Food and Drugs Control Center, Poland, was analyzed. The concentrations of Fe (32.22 ± 1.22–131.1 ± 3.26 mg/kg), As (ND to 0.12 ± 0.04 mg/kg), Cr (0.08 ± 0.01–3.2 ± 0.09 mg/kg), Pb (N.D - 0.21 ± 0.02 mg/kg) and Cd (ND to 0.14 ± 0.08 mg/kg) mg/kg were measured. The results revealed that the concentrations of Cr in all spice samples tested were much higher than the Maximum permissible limit (MPL) values. All spices in this study had THQ and HI values less than one, indicating that consumers will experience no potential health hazards from consuming specific metals through spices. However, continual scrutiny should be maintained.  •Spices are the dried parts of plants used historically to enhance food's flavor, color, and aroma. .•The species samples were digested using a microwave acid-assisted system.•5 heavy metals were determined simultaneously by ICP-MS.•ICP-MS demonstrated high sensitivity and confirmatory capacity needed for determining heavy metals.•The validated method was applied successfully to spice samples from South Africa. Spice roots, barks, fruits, and seeds are mostly used as ingredients in cooking processes to improve the smell, flavor, and color of food, as well as to hide unpleasant odors Spices and medicinal plants contain heavy metals that are essential to living cells. Understanding the content of trace metals in foods is critical because of their essential or hazardous effects on human health Spices are the dried parts of plants that have been used historically to enhance food's flavor, color, aroma, and acceptability Regarding heavy metal contamination, there is limited information available about the safety of spices, according to the literature, Pb was found in unbranded turmeric samples and was found to be above the permissible limit 22.13 was then added. According to the manufacturer's recommendations, the vessels were heated in a microwave system to a maximum of 1200 W before undergoing a microwave digestion program. Following thorough digestion, the clear solutions were poured into 50-mL volumetric flasks, filtered through Whatman No. 1 filter paper , and then made up to 50 mL using ultra-pure de-ionized water. The concentrations of Fe, Cd, As, Cr, and Pb were determined using ICP-MS NexION 300Q from PerkinElmer, USA. The analysis was carried out in accordance with the following procedure: sample introduction, fragmentation of compounds with a characteristic ion charge, nebulization, ionization of elements in high temperature argon plasma, detection based on m/z ratio, mass discrimination in the quadrupole analyzer, and data analysis as described in the A total of twenty common spices were randomly bought from registered shops in South Africa. The samples were kept in zip lock bags and were stored at room temperature, until analysis. The species samples were digested using a microwave acid-assisted system. 0.5 g of spice samples were weighed into Teflon vessels, and 10 mL of 65 % HNOPrior to the measurement, the ICP-MS spectrometer was calibrated using a multi-elemental mixture of metal standards . Each solution was measured three times, and the heavy metals were quantified using external calibration curves. The standard solution of equal concentrations of mixed heavy metals was diluted into seven different concentrations of 1 % nitric acid solution, and each standard solution received 50 μg/L of the internal standard (Sc). The analytical processes' quality was tested using Polish Certified Reference Material Mixed Herbs (INCT-TL-2) that were processed in the same way as the spice samples. A calibration solution was prepared using the ICP-MS multi-element stock standard solution (Merck) and internal standard (Sc) to acquire the sensitivity factors for the individual elements. The calibration curves for all elements show good linearity across the entire concentration range, with determination coefficients greater than 0.999. The limit of quantitation (LOQ) and limits of detection (LOD), as well as recovery was performed.2.2The potential health risks of consuming heavy metals through spices were assessed using the estimated daily intake (EDI), target hazard quotient (THQ), and Hazard Index (HI). The EDI value is determined by the amount consumed daily, body weight, and element concentrations in spices .(1)EDI=CEDI is estimated daily intake.metal (mg/kg) concentration of heavy metal content in spices,CIR (ingestion rate gram/day person) (10 g/day/person),BW represent the body weight (kg) (60 kg).THQ is used to assess the non-carcinogenic risks of long-term exposure to contaminants in Spices, and the calculations were done using To calculate the overall non-carcinogenic risk to human health posed by exposure to multiple pollutants, the HI was developed. HI is the sum of the HQ for all heavy metals found in spices, as shown in If the values of THQ/HI ≥ 1 imply that the population may have negative health effects, while if THQ/HI < 1, the population is not expected to encounter any obvious negative consequences 3The heavy metal content of various types of spices available in South African shops was measured in this study. The analytical data revealed that all of the samples contained a low concentration of Pb. Pb concentrations ranged from 0.08 ± 0.001 to 2.51 ± 0.01 mg/kg , these vCr highest mean concentration were found to be 3.2 ± 0.09 mg/kg and lowest mean concentrations were 0.08 ± 0.01 mg/kg as presented in The presence of As was found only in 2 samples (2 %) with the concentration ranges from N.D - 0.12 ± 0.04 mg/kg. The amount of the metal that was being measured in each of the samples that were analyzed fell below the permissible limit (0.2 mg/kg). Similar report of As was discovered ranging from 0.03 mg/kg and 0.06 mg/kg in nutmeg and black pepper samples to 0.37 mg/kg in oregano samples The concentration of Cd ranged between N.D to 0.14 ± 0.08 mg/kg while permissible limit of Cd is 0.2 mg/kg in plant Fe is a significant trace element, and mixtures of iron and proteins are essential for metabolism in all living things. According to analytical findings, the Fe content of spice samples ranged from 32 ± 1.22 to 131.1 ± 3.26 mg/kg . DespiteFe is a component of the active site of several reproductive hydrogenases and is most frequently linked to ligands that contain sulfur 3.1The EDI values of toxic metals through the consumption of spices are summarized in All spices had THQ values that were less than one, indicating that there is no potential health risk associated with consuming spices. 4In conclusion, the purpose of this study was to determine the amount of heavy metals present in some selected South African spices. Applying the optimized acid digestion method, which was validated through a recovery experiment, yielded good percentage recoveries (87–100 %) for the analysis of heavy metals in spices. All of the heavy metals investigated through the consumption of spice samples had HI values lower than 1. This shows that using these spices in different homes poses no significant non-carcinogenic health risk. Additionally, it is advised to discard the first infusion and draw from the second to reduce the amount of toxic metals that consumers consume in order to counteract the effects of the concentrations of toxic metals. Finally, it is proposed that heavy metals be continuously monitored in all food commodities in order to assess health risks from toxic heavy metals and safeguard consumers.O.M. Oladeji: Visualization, Project administration, Supervision, Validation, Data curation, Writing - original draft, Writing - review & editing. O.A. Aasa: Visualization, Software, Writing - review & editing. O.O. Adelusi: Visualization, Software, Methodology, Writing - review & editing. L.L. Mugivshisa: Project administration, Supervision, Validation, Funding acquisition, Writing - review & editing.The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.


X-linked adrenal hypoplasia congenita (AHC) is a rare disorder characterized by primary adrenal insufficiency (PAI) and hypogonadotropic hypogonadism (HH), with limited clinical and genetic characterization.The clinical, biochemical, genetic, therapeutic, and follow-up data of 42 patients diagnosed with X-linked AHC were retrospectively analysed.P < 0.05). The testicular volumes of the three patients with pulsatile gonadotropin-releasing hormone (GnRH) therapy were larger than those of the six patients undergoing hCG therapy (P < 0.05), and they also exhibited some growth in terms of luteinizing hormone (LH), follicle-stimulating hormone (FSH), and testosterone. Of the 42 patients, three had an Xp21 deletion, and 39 had an isolated DAX1 defect. Most patients (9/10) with entire DAX1 deletion accounting for 23.8% (10/42) of the total variants had early onset age of less than one year.Hyperpigmentation , vomiting/diarrhoea , failure to thrive , and convulsions  were the most common symptoms of X-linked AHC at onset. Increased adrenocorticotropic hormone (ACTH)  and decreased cortisol  were the most common laboratory findings, followed by hyponatremia  and hyperkalaemia . Thirty-one patients presented with PAI within the first year of life, and 11 presented after three years of age. Three of the thirteen patients over the age of 14 exhibited spontaneous pubertal development, and ten of them experienced delayed puberty due to HH. Six patients receiving human chorionic gonadotropin (hCG) therapy exhibited a slight increase in testicular size and had rising testosterone levels (both This study details the clinical features and genetic spectra of X-linked AHC. Patients with X-linked AHC show a bimodal distribution of the age of onset, with approximately 70% presenting within the first year of life. Pulsatile GnRH may be recommended for HH when hCG therapy is not satisfactory, although it is difficult to achieve normal testicular volume. The combination of clinical features and molecular tests provides information for an accurate diagnosis. DAX1 , which is also known as NR0B1 , with a prevalence of one in 70,000 to 600,000 males  in all patients (42/42), decreased cortisol levels [63.1 (21.7–139.3) nmol/L] in 88% of patients (37/42), decreased serum sodium levels (126.7 ± 8.9 mmol/L) in 76% of patients (32/42) and increased serum potassium levels (6.0 ± 1.4 mmol/L) in 69% (29/42) of patients (Table P > 0.05). In addition, a comparison of the biochemical parameters displayed no significant difference in the serum sodium and cortisol levels (both P > 0.05), but the serum potassium level of the infantile-onset group was higher than that of the childhood-onset group . This finding indicated that the degree of electrolyte disturbance in infantile-onset patients was more severe than that in childhood-onset patients with X-linked AHC.An apparent bimodal distribution pattern for age at presentation was observed in 42 patients with X-linked AHC. Of those, 31 cases (74%) presented within one year of age with a median age of 15.0 days (range: 0.0–1.0 year), while 11 cases (26%) presented over three years of age with a median age of 4.5 years (range: 3.0–8.2). Therefore, 42 patients were divided into the infantile-onset group (≤ 1 year) and the childhood-onset group (> 1 year) according to the onset age (Table 2/d and 0.05 (0.05–0.05) mg/d, respectively. Patients over 14 years of age (n = 13) had higher doses of HC than those under 14 years old (n = 29) . After the treatment, all the patients showed remission in clinical signs and symptoms of PAI, hyperpigmentation disappeared gradually, and the serum electrolyte levels returned to normal, although the ACTH level [305.0 (54.3–539.0) pg/ml] remained mildly high.The hydrocortisone (HC) and 9α-fludrocortisone replacement therapy was a crucial treatment for adrenal dysfunction in all patients, and the doses were 13.3 ± 2.9 mg/mAmong the 42 patients, thirteen patients were over 14 years of age and were assessed for the conditions of pubertal development Table . In thesP = 0.0160, 0.0070, 0.0140, 0.0028, respectively). Therefore, in this cohort, eleven of the 13 patients over 14 years old had HH, including ten with delayed puberty and one with spontaneous pubertal development followed by disrupted puberty.Spontaneous adolescent development was observed in three patients  during the follow-up at the age of 12.5 ± 0.3 years, with a testicular volume of 5.0 (5.0–5.5) ml, basal FSH level of 10.50 ± 1.35 IU/L, LH level of 0.98 ± 0.39 IU/L, and the testosterone level was 3.93 ± 1.61 nmol/L. The age of the last follow-up of these three patients was 15.6 ± 1.2 years old. Patient 2 was found to have pubertal arrest, with a decrease in testicular volume (from 6 to 3 ml) and testosterone level (from 5.79 nmol/L to 0.49 nmol/L) at 17.3 years old, suggesting arrested puberty. The peak LH and FSH levels after the GnRH stimulation test were 3.66 IU/L and 9.19 IU/L, respectively, suggesting the onset of HH. The other two patients (P9 and P10) had slow pubertal progression, and their testes were still around Tanner stage 2–3  two years after puberty initiation , and three received pulsatile GnRH administration . Of these three patients with pulsatile GnRH treatment, two patients (P20 and P32) had received hCG previously for 2.5 years and 6 months, respectively, but with limited effects. Thus, six patients had been treated with hCG. In addition, patient 2 recently underwent hCG therapy for less than one month, patient 21 received testosterone replacement, and the other two patients (P3 and P37) did not receive any management for HH. Therefore, a total of four patients  had no data on treatment outcomes.P = 0.0250). Their total testosterone levels increased from < 0.35 nmol/L in all but one patient  to 1.99 (1.20–2.91) nmol/L (P = 0.0313), which were still below the normal range for age in all patients except for one . The FSH and LH levels showed no change. This suggests that the increase in testicular volume and testosterone levels in the patients after hCG treatment was limited.Six patients who received hCG therapy for 6.5 (6.0–24.3) months exhibited an enlargement of testicles from 2.7 ± 0.4 ml to 3.3 ± 0.4 ml (P > 0.05), probably due to the small sample size.Three patients underwent pulsatile GnRH administration for 2.7 ± 1.5 months. A mild increase in testicular volumes [from 3.0 (3.0–3.3) ml to 4.5 (4.5–6.8) ml], basal LH levels [from 0.23 (0.12–0.31) IU/L to 1.77 (1.36–2.60) IU/L], basal FSH levels [from 2.84 ± 1.27 IU/L to 6.19 ± 2.39 IU/L], and testosterone levels [from all < 0.35 nmol/L to 1.43 (1.28–3.00) nmol/L] was observed  IU/L vs. 0.65 (0.23–0.77) IU/L, P = 0.0476]. Thus, pulsatile GnRH may be recommended for HH when hCG therapy is not satisfactory, although it is difficult to achieve normal testicular volume. Despite this, the height SDS of these 7 patients increased significantly after hCG medication and/or pulsatile GnRH therapy , possibly due to the elevated testosterone levels after treatment.We compared the final testicular volume, LH, FSH, and testosterone levels between the six patients receiving hCG therapy at the last follow-up (n = 4) or prior to the therapy alteration (n = 2) and in three patients undergoing pulsatile GnRH administration at the last follow-up (n = 3). The testicles and LH levels of the patients undergoing pulsatile GnRH were larger and higher than those undergoing hCG therapy [4.5 (4.5–6.8) ml vs. 3.3 ± 0.4 ml, Xp21 deletion syndromeIL1RAPL1 and DAX1 deletions (3.3 Mb) by CMA. He is currently 14.2 years of age, affected by intellectual disability and HH in addition to adrenal failure.Three patients  with Xp21 deletion syndrome were diagnosed by chromosomal microarray assays  through CMA. Unfortunately, he died at the age of six months due to an adrenal crisis.P38 presented with hyperpigmentation after birth and failure to thrive during the neonatal period. When he was six months of age, he manifested with delayed milestones (unable to lift his head) and hypotonia coupled with adrenal insufficiency. His two elder brothers had died at the ages of two months and four months. The laboratory findings showed that urinary glyceric acid level was elevated and creatine kinase in the blood was 5645 IU/L (reference range: 39–308 IU/L). Thus, Xp21 deletion syndrome was suspected by these clinical findings, and he was further diagnosed with IL1RAPL1, DAX1, GK, and DMD (3.6 Mb) deletions by CMA. He is currently four years old, speaks few words and is unable to walk due to intellectual disability and persistent hypotonia.P39 encountered hyperpigmentation after birth. When he was two years old, he presented with failure to thrive and hypotonia, with a length of 80 cm (< − 2SD) and a weight of 8.8 kg (< − 2SD). Biochemical findings showed that the serum creatine kinase level was 9431 U/L and that the urinary glyceric acid level was 653.16 (reference range: 0.0–20.0 mmol/mol creatinine). His developmental quotient (DQ) scores in the domains of adaptive behaviour, gross motor, fine motor, language, and social behaviour assessed by the Gesell Development Diagnosis Scale were 23, 23, 25, 35, and 37, respectively, suggesting severe developmental delay. He was further diagnosed definitively with 2.DAX1 variant spectrumDAX1 gene were identified in 42 patients, including missense (n = 6), nonsense (n = 6), frameshift (n = 14), and gross deletions (n = 4), which were all classified as pathogenic (P) or likely pathogenic (LP) . Fourteen variants were not reported previously [DAX1 gene, located at both the DBD and LBD domains, whereas missense variants clustered in the LBD domain  harboured a contiguous gene deletion. Among these three patients, one (P37) had a deletion of IL1RAPL1 and DAX1, and two (P38 and P39) carried the deletion extending distally to include the IL1RAPL1, DAX1, GK, and DMD loci.Of the 42 patients, 18 had frameshift variants, six had nonsense changes, and six had missense changes. The remainder  had a deletion of the locus containing 3.Genotype–phenotype analysisDAX1 alone presented with isolated X-linked AHC, while two patients (P38 and P39) at risk for the metabolic crisis or phenotypic characteristics of hypotonia and ID were found to have the deletion extended to contain the IL1RAPL1, DAX1, GK and DMD genes. A deletion extending to IL1RAPL1 and DAX1 was confirmed in one patient (P37) exhibiting ID in addition to adrenal dysfunction. Thus, a good genotype–phenotype correlation was confirmed in Xp21 deletion syndrome.First, nine patients with a deletion involving DAX1 deletion and three with Xp21 deletion, all except for one patient (P37) had an early onset age of less than one year, indicating that the entire DAX1 deletion might be associated with early onset age. In addition, no relation was found between the type of DAX1 variant and whether pubertal development can be initiated spontaneously.Then, an attempt was made to associate the severity of the disorder with specific variants. The age of onset of patients with X-linked AHC was used to assess the severity of adrenal insufficiency since it is the most readily available proxy. Of seven patients with isolated entire Finally, patients within a pedigree carrying the same variant may have different ages of onset. Patient 5A with the c.501del variant presented at one year of age, while his younger cousin (P5B) presented at 3.3 years of age. The age of onset of patient 8C with the c.676del variant was three years, while the age of onset of his younger cousins (P8A and P8B) was within the first five days and three days of life, respectively. It elucidated the phenotypic heterogeneity of X-linked AHC. In terms of phenotypic heterogeneity, genetic changes, epistatic interactions, and additional modifier variants may influence disease presentation in some single gene disorders. Although a group of modifier variants may influence the phenotype, not every modifier variant will be involved in every patient, implying a role for oligogenic inheritance in phenotypic expression , 30.X-linked AHC is a rare congenital disorder, but it still accounts for a substantial proportion of patients with primary adrenal insufficiency. To date, few studies have been reported on large cohorts of X-linked AHC patients. Previous studies mainly focused on describing the genetic composition without detailed clinical characteristics of X-linked AHC –34. The The proportion of DAX1 defects in PAI is currently unclear. Of the cases with PAI followed up in our centre over 20 years, 52.5% (42/80) of patients with biochemically uncharacterized PAI carried DAX1 defects (unpublished data). The biochemically uncharacterized PAI is PAI that cannot be diagnosed by specific biochemical findings or urine/serum steroid metabolites , excluding congenital adrenal hyperplasia, X-linked ALD, and autoimmune disease , 32. AccDAX1 deletion , including isolated DAX1 deletion (n = 7) and Xp21 deletion (n = 3). We compared the recurrent DAX1 variants in the Italian, British, and Japanese cohorts and found that the entire DAX1 deletion accounted for 25.0% (4/16), 16.7% (2/12), and 10.0% (2/20), respectively [DAX1 deletion may be the hotspot variant for X-linked AHC in these populations, accounting for approximately 10–25%. Furthermore, we compared the proportion of Xp21 deletion in X-linked AHC, which accounted for 25.0% (4/16) in Italy, 16.7% (2/12) in the United Kingdom, 7.1% (3/42) in our cohort, and 5.0% (1/20) in Japan [DAX1, GK, and DMD, three extended to include IL1RAPL1, DAX1, GK, and DMD (including our two patients), and one contained the locus of IL1RAPL1 and DAX1 (our patient). This suggests that most of the contiguous gene deletions in patients with DAX1-related X-linked AHC contain DAX1, GK, and DMD deletions. Therefore, we recommend that males with biochemically uncharacteristic PAI be first evaluated for serum creatine kinase, urinary glycerol acid, and mental development, followed by CNVs analysis to confirm the Xp21 deletion. In addition, the number of cases with Xp21 deletion in this study is small, and the long-term prognosis, including the progression of clinical features and the efficacy of comprehensive treatment, is still unknown. In our study, 26 DAX1 point variants were found in 27 non-consanguineous families, with only p.P335Tfs*55 discovered in two unrelated families. In reviewing genetic data from Turkish, Italian, British, and Japanese cohorts [DAX1 point variants were identified to occur in no more than two families , suggesting the occurrence of the recurrence of DAX1 point variants in unrelated families is a rare phenomenon [In DAX1, at least 200 distinct variants have been listed. The most prevalent variant in this study was the entire ectively –34. Thisin Japan –34. Of t cohorts –34, a feDAX1 deletion, and all but one of the ten patients harbouring the entire DAX1 deletion presented within the first year of life, suggesting that total DAX1 deletion may be linked to an earlier age of onset. Nevertheless, no difference was found in the frequency of other types of variants between the infantile-onset group and childhood-onset group, which was consistent with previous studies [The age of onset exhibits a bimodal distribution in X-linked AHC patients. The majority of patients 70–80%) presented in infancy, and the remainder presented insidiously later in childhood with an age range of two to nine years old , 7. Simi0–80% pre studies .DAX1 variant and whether pubertal development can be initiated spontaneously in our study. Spontaneous pubertal development is possible due to the presence of residual hypothalamic and pituitary function in these patients, making partial gonadotropin deficiency sufficient to allow spontaneous pubertal development up to mid-puberty [Most boys with X-linked AHC manifest HH as delayed puberty, whereas some boys can enter puberty spontaneously and progress to approximately Tanner stage 3 (or a testicular volume of 6–8 ml) between the ages of 15 and 18 years . In our -puberty , 19. TheDAX1 is expressed in the adrenal cortex, hypothalamus, pituitary, and testes. Previous studies revealed that HH in patients with DAX1 defects results from the combined deficiency of the hypothalamus and pituitary , 36, 37.DAX1 variant, PCR/Sanger sequencing is performed. If PCR results show no DAX1 amplification, indicating an isolated DAX1 deletion, qPCR must be used to confirm the gross deletion. Diagnoses and differential diagnoses can be made more quickly and affordably using this flowchart.The evaluation of clinical features is the initial and important step in determining the possible aetiology of X-linked AHC. In the current study, we clarified the diagnosis of 42 patients with X-linked AHC based on biochemical characteristics  and intellectual development assessment, along with genetic analysis. As a result, we suggest a flowchart for diagnosing X-linked AHC in children Fig. . First, DAX1 deletion appears to be related to an earlier age of onset. For pubertal development, the majority of boys with DAX1-related HH manifest with delayed puberty, while a small proportion of patients can spontaneously enter puberty followed by disruption. For the management of HH, all patients treated with either hCG or pulsatile GnRH had some degree of increase in testicular volumes and hormone concentrations but failed to reach normal levels. We propose a diagnostic process for children with X-linked AHC that will facilitate the rapid and comprehensive diagnosis of patients. A long-term follow-up and a larger sample size are needed to further evaluate prognosis and treatment.This study reported 42 patients with X-linked AHC, expanding our understanding of the clinical, biochemical, genetic, therapeutic, and follow-up characteristics of X-linked AHC. Patients with X-linked AHC exhibit a bimodal distribution of age of onset, with approximately 70% presenting within one year of age. The entire


Citrus reticulata Blanco peel extracts into wheat bread as a promising opportunity to increase their bioactive potential, along with supporting the sustainable management of citrus-industry waste. A control and a wheat bread enriched at 2% and 4% (w/v) with a phenolic extract from mandarin peels were prepared and analyzed for antioxidant activity and phenolic composition using LC-MS and UV-Vis spectrophotometry. In addition, in vitro digestion was performed, and the digested extracts were analyzed with HPLC-MS/MS. The results showed a significant increase in total flavonoid content , antioxidant activity (IC50 = 37 ± 4 mg·g−1), and contents of quercetin, caffeic acid, and hesperidin in the 4% (w/v) enriched bread. Yet, most polyphenols were completely degraded after the in vitro digestion process, barring hesperidin (159 ± 36 μg·g−1), highlighting the contribution of citrus enrichment in the development of an enriched bread with antioxidant potential.The fortification of foods with bioactive polyphenols aims to improve their functional properties and to provide health benefits. Yet, to exert their benefits, phenolic compounds must be released from the food matrix and absorbed by the small intestine after digestion, so assessing their bioaccessibility is crucial to determine their potential role. This work aims to incorporate  Over the last decade, consumers’ nutritional awareness has increased considerably around the world, leading to the search for healthier and more nutritious foods that ensure greater safety and promote well-being . As a reCitrus (Rutaceae) is the most widely produced tree fruit crop in the world, with more than 130 million tons per year. About 33% of citrus fruits are processed industrially for juice production, resulting in the generation of over 15 million tons of citrus waste per year, namely peels, membranes, and seeds, the management of which is a major economic and environmental concern − was chosen for online identification of polyphenols using mass detection in negative ion mode, where ESI capillary voltage was fixed at 3.5 kV. The drying gas temperature was set to 325 °C and a flowrate of 8.0 L min−1. The nebulizer pressure was fixed at 17 psi. Identification of phenolic compounds was carried out by comparing the retention times and molecular ions acquired from the standards with those of the sample extracts. External calibration curves were obtained for quantifying purposes. For on-column focusing purposes, injection solutions were prepared by adding to sample aliquots 50 μL of ACN, 800 μL of MeOH, 0.05% (v/v) aqueous formic acid solution at pH 2.9, and Milli-Q water to a final volume of 5 mL.Individual polyphenol determination of mandarin and wheat bread extracts was carried out using capillary liquid chromatography coupled to a mass simple quadrupole analyzer (cLC-ESI-MS), following the procedure previously described by Gómez-Mejía et al.  with minm/z range 100–1000, using a capillary voltage of 4 kV and a 45 psi pressure. Data processing was performed with Masshunter Data Acquisition B.05.00 and Masshunter Qualitative Analysis B.07.00. Nitrogen was used as both nebulizing and drying gas .For a more thorough and wider determination of the bioactive compounds present in the mandarin residues and in the samples subjected to the in vitro digestion process, high-performance liquid chromatography coupled to a quadrupole time-of-flight mass spectrometer (HPLC-ESI-QTOF) was employed. The phenolic analysis was performed in an Agilent liquid chromatography system (Mod. 1200), a quaternary pump (G1311A), a coupled degasser (G1322A), a thermostated automatic injector (G1367B), a thermostated column module (G1316A), and a QTOF mass spectrometer (Agilent G6530A) with atmospheric-pressure electrospray ionization source (ESI) and JetStream technology, operating in negative mode and scanning mode (SCAN) in the v/v) FA in ACN (solvent A) and 0.1% (v/v) FA aqueous solution (solvent B) as follows: 10% solvent B holding for 0.1 min, linear increase to 35% B within 30 min, and to 70% B within another 5 min. This condition was held for 2 min, then a final linear increase to 90% B was attained within 3 and was held for 5 min, subsequently followed by a re-equilibration of the column. The flow rate was set at 0.50 mL·min−1 and the injection volume was fixed at 20 μL. HPLC-QTOF separation was performed on a Synergi™ C18 Fusion-RP 80 Å analytical column , maintained at 30 °C and using a mobile phase gradient based on a mixture of 0.1% . LOD and LOQ were calculated for analyte concentration as 3 signal-to-noise (S/N) at height and 10 S/N, respectively. Finally, the precision was estimated at 80 μg·L−1 for gallic acid, dihydroxybenzoic acid, p-coumaric acid, resveratrol, quercetin, and kaempferol; 44 μg·L−1 for caffeic acid; 40 μg·L−1 for trans-ferulic acid; and 480 μg·L−1 for catechin. Intraday variation (n = 3) was assessed by injecting three standard solutions at the target concentration for each analyte on the same day, while inter-day precision was similarly obtained from three successive days . The relative standard deviation  was calculated for both the retention factor (k) and peak areas of each analyte.The performance of both LC-MS methods was evaluated for the standard solutions under optimal chromatographic conditions in terms of linear range and limits of detection (LOD) and quantification (LOQ) ,19. Linew/v) fortified and control wheat bread were subjected to an in vitro digestion process, according to the procedure described by Minekus et al. [−1), KSCN (40 g·L−1), NaH2PO4 (177.6 g·L−1), Na2SO4 (114.0 g·L−1), NaCl (350.6 g·L−1), NaHCO3 (169.4 g·L−1), and urea (50.0 g·L−1). The pH was set to 6.8 with 0.1 M HCl and incubated at 37 °C for 15 min. Subsequently, 5 mL of pepsin dissolved in 0.1 M HCl was combined, the pH was adjusted to 1.8 with 6 M HCl, and the gastric mixture was maintained at 37 °C for 2 h. The duodenal phase was then initiated by adjusting the pH to 6.8 with saturated aqueous NaHCO3 solution. Lastly, 3 mL of 1.5% (w/v) pancreatin and 0.15% (w/v) bile salts, both dissolved in 0.15 M NaCl, were added and the mixture was incubated at 37 °C for 2 h. Gastrointestinal digestion blanks were prepared in parallel. The procedure was performed in triplicate with constant shaking (100 rpm) and digested samples were analyzed fresh to prevent phenolic degradation, following the procedure described in The 4% , i.e., the percentage content of a polyphenol at the duodenal stage following the in vitro gastrointestinal step , was dett-Student test, one-way analysis of variance (ANOVA), and the least-significant difference (LSD) multiple comparison test using the software package Statgraphics 19 .Data were statistically analyzed using two-tailed paired p-coumaric acid, trans-ferulic acid, resveratrol, quercetin, and kaempferol, seeking the optimal chromatographic resolution in the minimum analysis time. Based on the LC separation proposed by Gómez-Mejía et al. [p-coumaric acid (0.5–180 μg·L−1), with R2 values above 0.9900. Further, acceptable LODs and LOQs were estimated in the range 0.1–6.0 μg·L−1 and 0.3–20 μg·L−1, respectively, apart from catechin (LOD = 100 μg·L−1), for which a lower sensitivity was observed, possibly associated with the negative ionization mode [A cLC-ESI-MS analytical method was developed, optimized, and validated for the determination of gallic acid, dihydroxybenzoic acid (DHB), catechin, caffeic acid, a et al. , the forion mode . In addi−1) and suitable R2 (≥0.9990) . Furthermore, the gastric phase showed a matrix inhibition below 70% by most of the phenolic compounds, while in the duodenal phase this effect was enhanced, especially for gallic acid, DHB, caffeic acid, and p-coumaric acid (>91%). Therefore, the quantification of the samples was carried out considering the inhibition percentages determined with Equation (1).To complement this analysis, an HPLC-ESI-QTOF method was also developed and validated to determine a larger number of phenolic compounds, including those reported to be abundant in citrus peels, i.e., hesperidin, naringin, and rutin ,23. Exac≥0.9990) . On the 7 μg·L−1 , respectv/v) mixtures, since they are considered efficient, GRAS, and environmentally friendly [Prior to the evaluation of the functionality of the enriched bread, an initial extraction step of phenolic compounds is required. Thus, to select the most suitable extraction method, different techniques were evaluated, namely, conventional SLE extraction  and UAE, using different equipment: an HD ultrasonic homogenizer with a 15.5 mm diameter probe and a Vibra-Cell homogenizer with a 2 mm tip. Conventional solvent extraction was selected because of its widespread use for phenolic extraction in a variety of agri-food matrices, including wheat bread ,25, was friendly .p-value < 0.05) in all determinations. As can be observed in −1) and antioxidant activity, i.e., the lowest IC50 (28 mg·g−1), followed by the UAE performed with the 15.5 mm probe. Positive correlations have been found between the TPC and the antioxidant activity of phenolic extracts [The extraction efficiency of each tested condition was expressed in terms of the TPC, TFC, and DPPH antioxidant activity and compared using ANOVA and LSD tests, showing significant differences . The better results of the probe in this respect could be due to the more efficient release of the matrix-bound flavonoids present in the wheat, such as rutin [50 value compared with the SLE  ethanol–water mixture was used, due to its greater effectiveness compared with extraction with water alone [p-value < 0.05) in all determinations. According to the data included in Citrus extracts were obtained following the SLE procedure described in er alone . In addier alone , so its er alone ,32. Analer alone ,15, and −1 and a TFC varying between 4 and 6 mg CE·g−1 when analyzing mandarin peel extracts (C. reticulata Blanco × C. Sinensis Osbeck) with EAU [−1 and TFC 22 ± 1 mg·g−1), demonstrating their potential as a source of high value-added compounds. Similarly, the antioxidant activity determined in this study was over 63 mg/100 g, exceeding the value reported herein (IC50 = 0.95 mg·g−1) for a mandarin peel citrus residue. The higher antioxidant activity of the present extract may be related to a greater content in phenolic compounds with significant free-radical-inhibition activity, such as gallic acid and quercetin [As a basis for comparison, Anticona et al. determined a TPC ranging from 6 to 9 mg GAE·gwith EAU , with thuercetin . Thus, tv/v) mixture. These sugars also react with the Folin–Ciocalteu reagent and may interfere with the determination and thereby alter the outcome, as previously stated [taralli  quadrupled from 0.09 mg QE·g−1 to 0.36 mg QE·g−1 when the dough was enriched with 17% (w/w) olive leaf extract [Citrus bergamia) pomace. In particular, a significant proportional increase in TFC was recorded when the citrus residue replaced from 2.5% to 15% (w/w) of the wheat flour in the formulation [w/v) enriched bread was quite substantial, with an IC50 reduction of more than tenfold (IC50 = 38 ± 2 mg·g−1) compared with the control bread (IC50 = 451 ± 40 mg·g−1), while the 2% (w/v) enriched bread halved its IC50 over the control  fortified bread . Several studies have reported the mentioned flavanone as the predominant phenolic compound in citrus peels, as such hesperidin has been determined in concentrations ranging from 1 to 15 mg·g−1 in Citrus reticulata Blanco [Citrus × clementina peels [−1, followed by quercetin and kaempferol, which were quantified at levels around 60 µg·g−1, and, lastly, by rutin and naringin, which were minor components  and p-coumaric acid (3.50 ± 0.07 µg·g−1); followed by the hydroxybenzoic acids; gallic acid (3.52 ± 0.01 µg·g−1); and dihydroxybenzoic acid, whose concentration did not exceed 1 µg·g−1  comparable to those in this study  compared with the control bread, while in the bread enriched at 4% (w/v), it was twelvefold higher (w/v) enriched bread to improve the total polyphenol content of wheat-based baking products. Specifically, the main effect of mandarin peel phenolic extract fortification was a significant increase in the flavonoid fraction, mainly focused on quercetin, myricetin, and naringin, which were not previously found in the control bread (−1 in the 4% (w/v) enriched bread and was the main phenolic compound in mandarin peel , probably due to the bonding of their free forms to the matrix components because of the baking process [p-coumaric acid, where a lower concentration  was observed for the enriched bread compared with the control. Likewise, Xiao et al. reported the loss of free kaempferol aglycone in a fermented buckwheat bread dough after steaming [p-coumaric acid in the enriched breads compared with the control could be attributed to the role of this polyphenol in the inhibition of acrylamide, according to the well-known Millard reaction, which is favored in the presence of reducing sugars, which are possibly co-extruded in the mandarin extract [p-coumaric acid most likely reacted with a precursor of acrylamide (3-oxopropanamide), giving rise to 2-(propenamide)coumaric acid [w/v) and 4%(w/v) enriched bread, with a concentration around 40 µg·g−1 in each. This could be related to the hydrolysis of the higher-molecular-weight dihydroxybenzoic acid derivatives, since the phenolic extract would have a lower pH than the EtOH–water mixture used in the control bread preparation, thus supporting their breakdown [w/w) sour orange albedo, compared with the control [As for the effect of bread enrichment, d higher . These rol bread , but alsrin peel . Likewis process . On the steaming . These fsteaming . On the  extract ,42,43. Xreakdown . Similar control . Therefop-coumaric, and trans-ferulic acids and the flavonoids rutin, hesperidin, and quercetin. Determining the content of phenolic compounds in foodstuff is not sufficient to predict their potential bioactivity. Indeed, it is well-known that the most widespread polyphenols in foods and beverages are not necessarily the most accessible, nor the most active, given that to perform their function, polyphenols must be released from the matrix, transferred into the bloodstream, and reach up to the target tissue or organ while undergoing mechanical, enzymatic, and metabolic transformations . Hence, w/v) enriched bread (7.1 ± 0.3 µg·g−1) compared with the control (1.71 ± 0.05 µg·g−1), demonstrating the effectiveness of fortification. p-Coumaric acid and rutin were determined in comparable concentrations in the salivary phases of both breads  enriched bread, as these compounds came directly from the mandarin extract , as well as of the flavonol quercetin, occurs in both bread samples, either due to acid hydrolysis caused by the low pH of the gastric medium or by their interaction with pepsin via non-covalent bonds [−1) (w/v) enriched bread  . This faed bread . In agrew/w) olive leaf extract, reporting such a significant decrease that the enrichment effect was suppressed [w/v) enriched bread, with double the duodenal concentration in the aforementioned case ((159 ± 36) µg·g−1 vs. (77 ± 5) µg·g−1 in the control). This fact is noteworthy, not only because it demonstrates the relevance and potential of mandarin-peel-enriched bakery products, but also because different models have shown that hesperidin consumption prevents lung damage, modulates antioxidant enzymes that induce apoptosis and cancer cell proliferation, and prevents cognitive deficits in cell lines, mice, and even in humans [After the duodenal phase, complete depletion of phenolic compounds occurs, except for hesperidin . A similppressed . Consequppressed ,45,46. Appressed ,48. Hespn humans . This suw/v). In addition, a study of the digestive stability of the polyphenols in the 4% (w/v) enriched bread was carried out and compared with the control bread. The results obtained showed a higher content of total individual polyphenols in the functionalized bread at all digestive stages, although, due to changes in the pH of the medium and digestive enzymes, only hesperidin could be quantified in the duodenal stage, with a concentration as high as 150 µg·g−1. Thus, such utilization of mandarin peels opens the way to an ecological and low-cost alternative to synthetic additives, with the aim of producing enriched foodstuff with potential health benefits for consumers.This study has proposed the valorization of citrus peel wastes by obtaining an extract rich in phenolic compounds, which was subsequently used in the preparation of an enriched bread. In particular, the effect of the enrichment percentage of phenolic extract has been evaluated in terms of antioxidant activity and phenolic content. A significant increase in the DPPH free-radical-inhibition potential and in the content of total flavonoids, in particular hesperidin, myricetin, and quercetin, has been observed to a greater extent in the bread enriched at 4% (


We have reported the relationship between low pulvinar nuclei (PN) intensity in susceptibility‐weighted imaging and the appearance of visual hallucinations and cognitive function. The aim of the study was to examine the changes in the quantitative susceptibility mapping (QSM) in patients with Parkinson's disease (PD) who underwent deep brain stimulation (DBS) and verify whether the PN susceptibility value (SV) on QSM can predict visual hallucination and cognitive changes after DBS.This study examined 24 patients with PD who underwent DBS along with QSM imaging on magnetic resonance imaging (MRI). All MRIs were performed within 3 months before surgery. The PN SV was further assessed based on the QSM. Then, associations were examined among cognitive changes, hallucination, and PN SV. The cognitive function of the patient was compared immediately before surgery and at 1 year postoperatively.p = .0037).Visual hallucinations were observed in seven patients during the follow‐up period. The PN SV was ≥0.045 ppm in nine patients with PD, and six of them had visual hallucinations, whereas only one of 15 patients with PD with SV of <0.045 ppm had visual hallucinations (Fisher's exact test, The SV of >0.045 ppm at the PN in QSM in patients with PD may provide useful information suggesting visual hallucination and cognitive deterioration after DBS treatment. In patients with Parkinson's disease 1 year after DBS, pulvinar nuclei in the QSM were associated with visual hallucination and cognitive worsening. We believe that this is useful information for the treatment and mechanisms of PD. Furthermore, we investigated the relationship between cognitive worsening and PN SV on QSM in these patients.22.1This study was conducted retrospectively. Inclusion criteria were as follows: patients with PD who fulfilled the United Kingdom Brain Bank criteria  at a 3T MRI unit  with a 32‐channel head coil. All MRIs were performed within 3 months before surgery. The pulse sequence for QSM was a gradient‐echo SWI sequence using the following parameters: repetition time (TR), 25 ms; echo time (TE), 21 ms; field of view (FOV), 230 mm × 206 mm; matrix size, 320 × 250; slice thickness, 1.6 mm; and acquisition time, 4 min and 2 s. In this study, we used research software provided by FUJIFILM Healthcare Corporation. This software method consists of three steps: (I) iterative least square minimization, (II) adaptive edge‐preserving filtering to the susceptibility map in the minimization process, and (III) weighted addition of the susceptibility map in k‐space before and after filtering. This process was described in previous reports  were implanted under local anesthesia using a Leksell stereotactic frame (Elekta Instruments AB) and anatomical (MRI and computed tomography) and physiological targeting. Electrodes were considered correctly located in the target region based on microelectrode recordings. Impulse generators  were implanted and connected during the second surgical procedure on the same day  of PN was set at the intercommissural (AC–PC) line slice. This ROI was determined in accordance with the Schaltenbrand and Wahren atlas , Hoehn–Yahr stage, LEDD, Mini‐Mental State Examination (MMSE), the Japanese version of the Montreal Cognitive Assessment (MoCA‐J), the Trail Making Test (TMT), Raven's colored progressive matrices (RCPM), the Center for Epidemiologic Studies Depression Scale (CES‐D), and Pareidolia Test, were performed for all patients before DBS  and at 1 year postoperatively . We performed multivariable analysis using a generalized linear model. As a predictor variable, we used the history of hallucination, the Put SV, and the PN SV. Receiver operating characteristic (ROC) curves were constructed to assess the sensitivity and specificity of the pulvinar or putamen SV in predicting postoperative hallucination. Cohen's d was calculated by G*Power . The Wilcoxon signed‐rank tests were used to compare the Hoehn–Yahr stage, MDS‐UPDRS, MMSE, MoCA‐J, RCPM, Pareidolia test, and CES‐D between baseline and 1 year postoperatively. Additionally, paired 3p = .048, .002, respectively). Other cognitive performance tests remained unchanged  was 62.7 ± 8.7 years. Additionally, the average motor performance of off state (Hoehn–Yahr stage and MDS‐UPDRS part III) significantly improved and LEDD and L‐dopa dosage significantly decreased 1 year postoperatively  were determined in seven cases but not 17 cases (H−). The type of hallucination was both visual and auditory in only one case and only visual in six cases. In only one case, hallucinations persisted 6 months after DBS. In the other six patients, hallucinations were transient, occurring three times in one case and only once in all other cases. At the time of preoperative evaluation, no patient experienced hallucinations. Among seven cases with postoperative hallucinations, five had a history of hallucination ; among the other 17 cases, only two had a history of hallucination. Cohen's t‐test, t (22) = −2.960, p = .007; t (22) = −2.640, 0.015, respectively). No significant difference was found in the QSM image SV in other nuclei between the H+ and H− groups    Table .p < .001), 1.006 , and 1.009 , respectively  at the PN SV and 0.80  at the Put SV. Using ROC curves to predict preoperative hallucination, the AUC was 0.60  at the PN SV and 0.62  at the Put SV  was 0.82 . A significant difference was also found in RCPM worsening over 1 year at 2.9 ± 2.5 in the SV of >0.045 ppm and 0.0 ± 3.1 in <0.045 ppm (p = .025). No other significant differences were observed in the assessment of MDS‐UPDRS, Hoehn–Yahr stage, and cognition , whereas the inter‐rater ICC was 0.68, 0.89, 0.86, 0.91, 0.90, 0.73, and 0.73, respectively .The SV analyses revealed the intra‐rater ICC for the SN, RN, STN, Put, GP, PN, and Cau as 0.78, 0.89, 0.80, 0.93, 0.93, 0.88, and 0.93, respectively (95% CI: 0.56−0.90, 0.76−0.95, 0.59−0.91, 0.86−0.97, 0.84−0.97, 0.74−0.94, and 0.84−0.97, respectively, all 4This study revealed that the SV of PN and Put in QSM was associated with experiencing visual hallucinations during the first year after DBS surgery. Additionally, a relationship was found between PN SV and RCPM worsening. These results are similar to our previous study (Matsuura et al., Recently, the relationship between brain iron deposition assessed by QSM and cognitive severity in PD was reported. The lower MoCA score revealed increased brain tissue iron in the hippocampus, thalamus, Put, and caudate nucleus (Thomas et al., However, the appearance of postoperative hallucinations was related to the SV of PN and Put and the history of preoperative hallucinations in the multivariable analysis. More factors need to be examined to predict the occurrence of hallucinations more accurately, and further studies with larger numbers of cases are needed.Seed‐based analyses in patients with DLB with visual hallucination reported decreased connectivity of the lateral geniculate nucleus, the superior parietal lobule, and the Put with the medial frontal gyrus (Pezzoli et al., As mentioned above, caudate nuclei have also been reported to be associated with cognitive function. In the present study, however, no significant changes were found in the occurrence of visual hallucinations. However, there was a slightly higher tendency in the hallucination occurrence group, so the results may not have been clear due to the insufficient number of cases.The results of this study indicate a significant improvement in MMSE scores obtained after 1 year of receiving DBS treatment compared with those obtained preoperatively. As PD is accompanied by fluctuating symptoms, preoperative evaluations are often performed when patients are in their ON state. However, due to the unpredictable nature of these symptoms, it is not always possible to evaluate optimal conditions. Notably, research has shown that patients experience slowness of thinking in their OFF state, suggesting that these effects may still influence the evaluation outcomes (Witjas et al., The present study had several limitations. First, the ROIs of PN and other targets were assessed freehand. Future studies should confirm and extend our findings using the auto‐segmented method. Second, this study has a small sample size, and the follow‐up period is not long enough. Therefore, further study is needed. Third, hallucinations were evaluated only by conducting inpatient and outpatient interviews. In the future, it will be necessary to use well‐validated evaluation criteria such as Neuropsychiatric Inventory (Cummings et al., In conclusion, the SV of ≥0.045 ppm at the PN in patients with PD may provide information that is useful for suggesting visual hallucination and cognitive deterioration after DBS treatment.Keita Matsuura, Yuichiro Ii, Masayuki Maeda, Akihiro Shindo, and Hidekazu Tomimoto helped with the conception of the study. Keita Matsuura, Hiroyuki Kajikawa, and Naoko Nakamura helped with the organization of the study. Keita Matsuura, Tommohiro Araki, and Hiroyuki Kajikawa executed the study. In terms of statistical analyses, Ken‐ichi Tabei and Keita Matsuura designed, Ken‐ichi Tabei executed, and Hiroyuki Kajikawa and Masayuki Satoh helped with the review and critique. In terms of QSM analyses, Keita Matsuura and Masayuki Maeda designed; Maki Umino, Keita Matsuura, and Ken‐ichi Tabei executed; and Masayuki Maeda helped with the review and critique. Keita Matsuura drafted the first manuscript, and Yuichiro Ii, Masayuki Maeda, and Hidekazu Tomimoto helped with the review and critique.Ken‐ichi Tabei and Masayuki Satoh are employed by Tokyo Metropolitan Public University Corporation. The remaining authors declare no conflicts of interest.https://publons.com/publon/10.1002/brb3.3263.The peer review history for this article is available at


IL7R deficiency is a rare form of SCID that usually presents in the first months of life with severe and opportunistic infections, failure to thrive, and a high risk of mortality unless treated. Although recent improvements in early diagnosis have been achieved through newborn screening, few IL7R-related SCID patients had been reported in the Chinese population.Severe combined immunodeficiency (SCID) is a group of fatal primary immunodeficiencies characterized by the severe impairment of T-cell differentiation. IL7R were inherited from the father and mother.Here, we retrospectively analyzed a case of SCID in a 5-month-old girl with symptoms, including severe T-cell depletion, recurrent fever, oral ulcers, pneumonia, hepatosplenomegaly, bone marrow hemophagocytosis, and bacterial and viral infections. Whole-exome sequencing (WES), quantitative PCR (qPCR), and chromosome microarray analysis (CMA) were performed to identify the patient’s genetic etiology. We identified a 268 kb deletion and a splicing variant, c.221 + 1G > A, in the proband. These two variants of IL7R gene deletion in combination with a pathogenic splicing variant in a patient with SCID. This deletion also expands the pathogenic variation spectrum of SCID caused by IL7R. The incorporation of exome-based copy number variant analysis makes WES a powerful molecular diagnostic technique for the clinical diagnosis of pediatric patients.To our knowledge, this is the first report of whole  IL7R gene abolish T cell development and function, resulting in SCID. IL7R deficiency causes T–B+NK+ SCID, which is responsible for 10% of the SCID cases [IL7R deficiency have been reported worldwide since it was first described by Puel et al. in 1998 [Severe combined immunodeficiency (SCID) is a life-threatening condition leading to early infant death as a result of severe infection, due to impaired T lymphocyte differentiation , 2. The ID cases –11. Over in 1998 –23.IL7R gene is located at 5p13.2 and consists of two subunits, the IL7R alpha chain (IL7Rα) and common gamma chain (γc). It is expressed on lymphoid cells and plays an important role in the development, survival, homeostasis, and proliferation of T cells [IL7R, with a maternal splicing variant, c.221 + 1G > A, and a paternal 268 kb deletion at 5p13.2 in a Chinese patient with SCID.The human  T cells , 25. Cur T cells , 27. Her9/L, with 88% neutrophils and 6% lymphocytes, a low hemoglobin level (8.6 g/dL), a high platelet count (815 × 109/L), an elevated level of ferritin (732.5 ng/mL) and bilirubin (57.1 µmol/L), presence of Epstein-Barr virus and Rotavirus, and a positive result for direct anti-human globulin test. Additionally, bone marrow examination revealed active hemophagocytosis  was deceased in May 2011, but there were stored dried blood spots. The proband was a girl, the second daughter of non-consanguineous parents. She was a product of a full-term pregnancy via cesarean delivery, and her birth weight was 3.05 kg. She presented at age 3 months with yellow discoloration of the skin and sclera, tan urine, recurrent fever, oral ulcers, and pneumonia. Physical examination revealed that the patient’s spleen was 2 cm below the left costal margin in the midclavicular line with a soft and sharp margin, and the liver was 2.5 cm below the right costal margin in the mid-clavicular line. Laboratory examinations revealed a white blood cell count of 10.6 × 10Dried blood spots were obtained from the proband, and blood samples were obtained from the unaffected parents and siblings after obtaining informed consent. Genomic DNA was extracted from dried blood spots and peripheral blood leukocytes for WES using SureSelect Human All Exon V6 kits  and a HiSeq2500 sequencer . The pathogenic variants were assessed using a protocol issued by the American College of Medical Genetics and Genomics (ACMG) . Co-segrIL7R (NM_002185.5), c.221 + 1G > A, which was related to SCID and was previously reported in a Chinese infant [IL7R gene using WES. Notably, the results suggested that the proband, her father, and sister may have a heterozygous deletion of the IL7R gene (NM_002185.5: chr5:35700839-35968226del). To further confirm the deletion of the IL7R gene, we performed qPCR. qPCR was performed to amplify IL7R exon 1, exon 4 and 8-noncoding region boundaries with three primers sets . All the experiments were replicated three times to ensure the accuracy of the experiment. After qPCR was performed, the copy number data were collected and analyzed using the 2−ΔΔCT method. The qPCR results showed that the patient, her father, and her sister had approximately half of copy numbers for exons 1, 4, and 8-noncoding region boundaries of IL7R compared to those in the control, suggesting that one of the IL7R alleles was entirely deleted. To examine the IL7R heterozygous deletion, we performed chromosome microarray analysis (CMA) on the father and sister (the proband did not have enough DNA samples for CMA)  had a novel homozygous variant in e infant . We excl7R NM_00285.5, c.2 had a no7R NM_00285.5, c.2IL7R gene combined with a splicing variant, (c.221 + 1G > A), as the cause of the SCID phenotype in a female patient. The deletion was paternally inherited and the c.221 + 1G > A variant was maternally inherited.Currently, the combination of pathogenic variant screening and CNV calling has been increasingly used in WES, which largely improves the detection efficiency for the diagnosis of inherited diseases , 31. RecIL7R associated with SCID in the ClinVar database included 14 missense, 10 nonsense, seven frameshift, six splicing variants, and one synonymous variant , and exon 3 is a deletion hotspot in IL7R [IL7R gene in a patient with SCID, emphasizing the importance of identifying CNVs of IL7R in such cases. Furthermore, the 268 kb deletion in the patient also involved three genes adjacent to IL7R, SPEF2, CAPSL and UGT3A1, the heterozygous deletion of which did not cause a dominant genetic disorder.The  in IL7R , 21. NotIL7R function may have immune dysregulation, severe infections, chronic inflammatory diseases, as well as cancer [IL7R. The atypical symptoms presented by our patient highlight the diagnostic challenges in the field of SCID. Typically, patients with complete loss of IL-7R present early onset SCID with profound T-cell lymphopenia and normal B and NK cell levels [IL7R allele in a Chinese family with SCID. We confirmed that the pathogenic variant c.221 + 1G > A in one IL7R allele is revealed by a 268 kb 5p13.2 deletion of the other allele. The simultaneous detection of SNVs/indels and CNVs in IL7R demonstrates the advantage of WES for identifying pathogenic variants and the importance of parental verification.Patients with SCID and impaired s cancer . The mail levels . UnfortuIL7R gene with a novel 268 kb deletion and a splice variant. This deletion also expands the pathogenic variation spectrum of SCID caused by IL7R. Further functional validation is necessary to clarify the pathogenesis of the IL7R gene in SCID.In conclusion, our patient received a definitive genetic diagnosis of SCID after her death, which was due to a compound heterozygous for the


In the published article, there was an error. In the Abstract, there were errors in the reporting of beta values and corresponding confidence intervals. The Results section of the abstract previously stated, “Results: Results supported our hypothesis that the association between romantic conflict and gambling-related problems would be sequentially mediated through negative affect and coping gambling motives, β = 0.38, 95% CI , and also showed a strong single mediation pathway through negative affect alone, β = 0.27, 95% CI .”The corrected section appears below:Results“Results supported our hypothesis that the model would explain a significant amount of variance in gambling-related problems, β = 0.35, 95% CI , and that the association between romantic conflict and gambling-related problems would be sequentially mediated through negative affect and coping gambling motives, β = 0.07, 95% CI , and also showed a strong single mediation pathway through negative affect alone, β = 0.24, 95% CI .”The authors apologize for this error and state that this does not change the scientific conclusions of the article in any way. The original article has been updated.


This randomized controlled trial was performed to observe the effect of remimazolam with sufentanil on hemodynamics during anesthetic induction in elderly patients with mild hypertension undergoing orthopedic surgery of the lower limbs.Sixty elderly patients were randomly assigned to undergo general anesthesia with intravenous injection of either remimazolam besylate  at 0.2 mg/kg  or propofol at 1.5 mg/kg . Both injections were completed within 15 to 20 s. If the bispectral index value did not reach 40 to 60, then 0.05 mg/kg of remimazolam was added in Group P and 1 mg/kg of propofol was added in Group R. When the BIS value reached 40 to 60, sufentanil was administered at 0.3 to 0.5 µg/kg and cisatracurium was administered at 0.15 to 0.2 mg/kg in both groups. Three minutes later, tracheal intubation and controlled ventilation were performed to maintain the end-tidal carbon dioxide partial pressure at 4.5 to 5.0 kPa. The mean arterial pressure (MAP), heart rate (HR), cardiac output (CO), continuous cardiac index (CI), systemic vascular resistance (SVR), and pulse oxygen saturation were recorded before induction (T0), when the eyelash reflex disappeared (T1), immediately after endotracheal intubation (T2), 1 min after endotracheal intubation (T3), and 5 min after endotracheal intubation (T4). The disappearance time of the eyelash reflex, injection pain, hypotension, bradycardia, hiccupping, nausea and vomiting, and other adverse events were observed.The MAP, HR, CO, and CI at T1, T2, T3, and T4 were significantly higher in Group R than P, while SVR was significantly lower in Group R than P (P < 0.05). In Group P, the MAP, HR, CO, and CI were significantly lower and the SVR was significantly higher at T1, T2, T3, and T4 than at T0 (P < 0.05). Adverse events occurred in 8 (20%) patients in Group R and 22 (73%) in Group P. The total incidence of adverse events was significantly lower in Group R than P (P < 0 0.001).Remimazolam combined with sufentanil for general anesthesia induction has the advantages of small hemodynamic fluctuations, stable circulation, and few adverse reactions, making it suitable for elderly patients with mild hypertension.Chinese Clinical Trial Registry . With the aging of the population, increasing numbers of elderly patients need surgical treatment. Hypertension is very common in elderly patients. Because elderly patients with hypertension generally have poor vascular elasticity, declining cardiovascular reserve function, and weakening of compensatory ability, hemodynamic fluctuations are likely to occur during anesthetic induction in these patients, potentially inducing serious complications. Propofol is a widely used sedative drug in the clinical setting and has the advantages of a short half-life and rapid recovery. However, the use of conventional doses of propofol in elderly patients is associated with a risk of complications such as hypotension, bradycardia, and arrhythmia . TherefoRemimazolam is a new type of anesthetic that has raised widespread concern. Remimazolam is an ultra-short-acting intravenous benzodiazepine sedative/anesthetic with the pharmacological properties of rapid onset, rapid recovery, and relatively small effects on hemodynamics . CompareThis study was approved by the Medical Ethics Committee of The Sixth Affiliated Hospital of Wenzhou Medical University in Lishui, China (LLW-FO-403) and was registered at the Chinese Clinical Trial Registry . Written informed consent was obtained from all patients. All methods were carried out in accordance with relevant guidelines and regulations.After obtaining approval from the hospital’s medical ethics committee and written consent from the patients, we enrolled 60 elderly patients with mild hypertension who underwent elective orthopedic surgery of the lower limbs from August 2021 to March 2022. The selected patients met the following criteria: 65 to 75 years of age, American Society of Anesthesiologists physical status of II or III, cardiac function class I or II, and blood pressure controlled within 160/90 mmHg after standardized treatment for those with a history of essential hypertension. None of the patients had serious arrhythmia or serious liver and kidney dysfunction, and none had a history of mental illness. The exclusion criteria were hypertension without regular medical therapy, bradycardia, and second-degree or worse atrioventricular block. The patients enrolled in the study comprised 27 male and 33 female patients with an age of 65 to 80 years and body weight of 50 to 78 kg. According to the order of treatment, the patients were randomly divided into two groups of 30 patients each: remimazolam 0.2 mg/kg (Group R) and propofol 1.5 mg/kg (Group P) , and bispectral index (BIS) value were continuously monitored with a multifunctional monitoring device . Upon completion of monitoring, a noninvasive continuous arterial blood pressure and hemodynamic monitoring system  was used to monitor the patients’ hemodynamic indexes, and efforts were made to relieve any nervousness. The baseline HR and MAP were defined as the average of three HR and MAP measurements before the procedures. All patients were given oxygen via a face mask (6 L/min) before the induction of anesthesia. Remimazolam besylate  was intravenously administered at 0.2 mg/kg in Group R, and propofol  was intravenously administered at 1.5 mg/kg in Group P. Both injections were completed within 15 to 20 s. If the BIS value did not reach the range of 40 to 60, then 0.05 mg/kg of remimazolam was added in Group P and 1 mg/kg of propofol was added in Group R. In both groups, controlled ventilation was performed after the eyelash reflex disappeared. When the BIS value reached 40 to 60, sufentanil was administered at 0.3 to 0.5 µg/kg  and cisatracurium was administered at 0.15 to 0.2 mg/kg in both groups. Three minutes later, tracheal intubation and controlled ventilation were performed to maintain the end-tidal carbon dioxide partial pressure at 4.5 to 5.0 kPa. Anesthesia was maintained with propofol at 3 to 5 mg/kg/h, remifentanil at 10 to 15 µg/kg/h, and 1–2% sevoflurane; cisatracurium was intermittently injected, and the BIS value remained at 40 to 60. Sevoflurane, propofol, and remifentanil were discontinued during skin suturing. The disappearance time of the eyelash reflex and hemodynamic indices including MAP, HR, cardiac output (CO), continuous cardiac index (CI), and systemic vascular resistance (SVR) were measured at five different time points: before induction (T0), when the eyelash reflex disappeared (T1), immediately after endotracheal intubation (T2), 1 min after endotracheal intubation (T3), and 5 min after endotracheal intubation (T4).The patients in both groups were routinely administered 10 mL/kg/h lactated Ringer’s solution in a readily accessible hand vein upon admission, and the mean arterial pressure (MAP), electrocardiogram, heart rate (HR), pulse oxygen saturation . In our pilot study, the incidence of hypotension elicited by 1.5 mg/kg of propofol was 40% (8/20), which was reduced to 10% (2/20) after pretreatment with 0.2 mg/kg of remimazolam. To achieve 80% (β = 0.20) statistical power with α = 0.05, each group would require no less than 23 patients. To account for drop-outs, we recruited 30 patients per group.Data processing was performed using SPSS 15.0 . The normality test in SPSS was used to determine whether the data conformed to a normal distribution. Measurement data are expressed as mean ± standard deviation. Repeated-measures analysis of variance was performed for intragroup comparisons, and one-way analysis of variance was performed for intergroup comparisons. A P value of < 0.05 was considered statistically significant.This trial was conducted from August 2021 to March 2022. Sixty eligible patients were screened and included in the final analyses. The patients’ demographic characteristics, including sex, age, and body mass index, were similar between the two groups (P > 0.05) . The MAP, HR, CO, and CI at T1, T2, T3, and T4 were significantly higher in Group R than P, while SVR was significantly lower in Group R than P. In Group P, the MAP, HR, CO, and CI at T1, T2, T3, and T4 were significantly lower than those at T0, while the SVR at T1, T2, T3, and T4 was significantly higher than that at T0 (P < 0.05)  patients in Group R and 22 73%) in Group P. The total incidence of adverse events was significantly lower in Group R than P (P < 0.001). There was no significant difference in hiccupping or nausea and vomiting between the two groups (P > 0.05) (Table % in GrouIn this study, we demonstrated that the MAP, HR, CO, and CI at T1, T2, T3, and T4 were significantly higher in Group R than P, while SVR was significantly lower in Group R than P. The incidence of total adverse events was 73% in patients in Group P but only 20% in patients in Group R.Elderly patients’ health is often complicated by a variety of diseases, of which hypertension is the most common. Their tolerance to anesthesia and surgery is lower than that of younger patients. Severe hemodynamic fluctuations can easily induce cerebrovascular accidents. Tracheal intubation during anesthetic induction is a severely noxious stimulus and can have a severe impact on patients’ hemodynamics. Maintaining stable hemodynamics of elderly patients during induction and improving the blood perfusion of the heart, brain, kidney, and other important organs can improve the disease outcome and prognosis of elderly patients , 8. How As a commonly used anesthetic in the clinical setting, propofol has the advantages of rapid onset, short duration of action, no accumulation in the body, and minimal postoperative nausea and vomiting. Propofol is widely used in the induction and maintenance of anesthesia. It exerts its sedative and hypnotic effects through interaction with gamma-aminobutyric acid receptors and has certain cardiovascular inhibitory effects that can lead to a decrease in blood pressure, CO, and HR –11; thesRemimazolam is a new benzodiazepine drug that has the advantages of rapid recovery and metabolism, no drug accumulation, independence from liver and kidney metabolism, and few side effects –19. HoweLike propofol, remimazolam can also be used for anesthetic induction in elderly patients. The injection pain was significantly less severe with remimazolam than with propofol, thus improving patients’ comfort. Remimazolam combined with sufentanil for general anesthesia induction produces minimal hemodynamic fluctuation, stable circulation, and few adverse reactions, making it suitable for elderly patients with mild hypertension.


So it’s imperative to select high carbon sequestering landscape tree species while considering their aesthetic values of urban green space.Continuous increasing carbon dioxide (COwCO2), daily net carbon sequestration per unit land area (WCO2) and daily net carbon sequestration of the whole plant (QCO2) through cluster analysis. Besides that, we found out the key factors affecting carbon sequestration potential of landscape tree species by redundancy analysis.32 tree species were selected as test objects which were commonly used in landscaping in Zhengzhou, a typical northern city of China. To assess the comprehensive carbon sequestration potential of landscape tree species in different plant configuration types, we simultaneously considered their daily net carbon sequestration per unit leaf area , V odoratissimum, P. Serratifolia, S. oblata among small arbors (SA), and B sinica var. Parvifolia, B. Megistophylla, L quihoui among shrubs (S) were recommended for local urban green space management. Photosynthetic rate (Pn), crown area (CA) and leaf area index (LAI) were the key factors which affected the comprehensive carbon sequestration potential both for LA, SA and S. Countstrategy . Urban amissions , 9, espe biomass –12. For settings , 14. Howsettings , 16.Previous studies have shown that green space in and around urban settings can contribute to ecosystem services and affect the sustainability of urban ecosystem as well as the livability of urban dwellers , 18, so Various methods have been used to quantify the amount of plants carbon sequestration capacity, such as biomass method, assimilation method, eddy covariance method, model simulation method and remote sensing method, etc. –37. The Zhengzhou City is located in northern China with warm temperate continental monsoon climate, and the common landscape tree species here have the typical characteristics of landscape vegetation configuration in this climatic zone . There aThe main goals of this study are as follows: Firstly, evaluate the comprehensive carbon sequestration potential of 32 common landscape tree species; Secondly, assess the carbon sequestration potential of different plant configuration types; Thirdly, find out the key factors that affect carbon sequestration potential of landscape tree species. Our results are expected to provide a scientific basis for rational allocation of landscape tree species and quantitative evaluation of environmental benefits.2, of which the built-up area of central urban area is 744.15 km2, and the urbanization rate is as high as 79.1%. Terrain is generally high in the southwest and low in the northeast. Also, it is characterized by four distinct seasons with temperate continental monsoon climate. Average annual temperature here is 14.7°, with the lowest temperature in January and the highest temperature in July. Besides, average annual rainfall is 632.4 mm, mainly from June to August, and average annual frost free period is 212.6 days. Soil type in this area belongs to brown soil and cinnamon soil, zonal vegetation belongs to temperate deciduous evergreen mixed broad-leaved forest belt, and flora belongs to north central temperate distribution and east Asia distribution.Zhengzhou  is located in the north–south transition zone of China, which has rich tree species . It is a4 m2. All trees grew at a relatively consistent climatic environment and management conditions. Also, the age of all trees was 10–15 years, and they were all healthy plants without obvious diseases and pests. Our study was carried out in the sunny weather which had sufficient natural light source without wind and rain from July to September in 2022 when plants grew vigorously. The average air temperature, relative humidity, duration of sunlight, wind speed and total solar radiation was 27.0 ℃, 68.2%, 5.3 h d−1, 9.5 km h−1 and 18185.7 kJ m−2 d−1, respectively, and the total rainfall was 224.2 mm during the investigation period.32 common landscape tree species were selected in this test through a comprehensive survey in the urban area of Zhengzhou, including of 17 large arbors , 8 small arbors  and 7 shrubs (4 deciduous shrubs and 3 evergreen shrubs) (Table LAI) were selected in our studies for their potential relationship with trees carbon sequestration capacity according to previous studies [Photosynthetic indexes, growth indexes, physiological indexes and Leaf area index  values were recorded for each leaf, and the average value was finally taken. Meanwhile, transpiration rate (Er), stomatal conductance (Ci), intercellular CO2 concentration (Gs), etc., were recorded.Determination of photosynthetic indexes: The LCpro SD portable photosynthetic instrument  was used to measure the photosynthetic physiological and ecological indicators of the tested tree species. Three trees with similar growth vigor were selected for all tested tree species, and five undamaged, well grown and mature leaves were selected for each tree for determination. The sampling work started at 8:00 and ended at 18:00, and the measurement was conducted every 2 h. Measurement work started when the instrument system was stable. Six instantaneous photosynthetic rate (DBH) of all tested trees, and laser rangefinder was used to measure their height (H) and crown diameter (CD). Besides, crown area (CA) was calculated by estimation method, which was the result of product of the east–west and north–south crown diameter.Determination of growth indexes: girth was used to measure the diameter at breast height (CHl): select 15 mature and fully developed leaves, wipe the surface of the leaves with a paper towel and waite for measurement. Used a portable chlorophyll meter (SPAD-502 PLUS) to measure the chlorophyll content of the leaves. Avoided the veins and petioles when measuring, and finally taken the average value.Determination of physiological indexes: Chlorophyll content (LAI): take canopy photos with a digital camera which connected to a fisheye lens at a height of 1.65 m above the tested tree species, and use Gap Light Analyzer (GLA) Version 2.0 to identify and analyze the clear photos that were easy to distinguish the sky and the canopy, and then get the leaf area index after sorting.Leaf area index , and pi is the instantaneous cooperative utilization rate at initial measurement point of tree species (μmol m−2 s−1), pi+1 is the instantaneous cooperative utilization rate of tree species at i + 1 measuring point(μmol m−2 s−1), ti is the instantaneous time (h) of the initial measuring point of the tree species, ti+1 is the instantaneous time (h) of the tree species at the measuring point i + 1, n is the number of tests, q is the daily average photosynthetic rate (μmol m−2 s−1), 3600 represents 3600 s per hour, and 1000 represents 1000 μmol per 1 mmol.Photosynthetic carbon sequestration index: based on Han Huanjin’s calculation principle of daily assimilation amount of photosynthesis, the net assimilation amount of plants on the day of measurement was used to estimate the amount of carbon sequestration of plants .1\docume2 + 4H2O → CH2O + 3H2O + O2. And daily net carbon sequestration amount was converted by daily net assimilation amount.The amount of plants carbon sequestration was calculated according to the reaction equation of photosynthesis: COwCO2):Daily net carbon sequestration per unit leaf area (WCO2):Daily net carbon sequestration per unit land area (QCO2):−2 d−1). The night respiration consumption of the tested tree species was calculated as 20% of the total net assimilation amount of the tree species in the day [2. −2 d−1), −1), LAI is the leaf area index, and CA is the crown area (m2).Daily net carbon sequestration of the whole plant ( the day . 44 is tCV)CV is the coefficient of variation. According to Wilding’s classification of variation degree, it indicates weak variation when CV ≤ 15%, and indicates medium variation when 16 ≤ CV ≤ 35%, and indicates strong variation when CV ≥ 36% [Coefficient of variation , Canoco 5  and Origin 2021  were used for all statistical analyses. The comprehensive carbon sequestration potential was assessed with cluster analysis method by R packages “cluster” . And theDBH, H and CA for all types, which were generally shown as LA (large arbors) > SA  > S (shrubs). In addition, DBH, H and CA of each tree species in a same plant configuration types also differed considerably. Specifically, DBH (34.3% < CV < 60.2%) and H (25.5% < CV < 42.9%) both showed moderate or strong variability, but except for deciduous small arbors  and evergreen small arbors , respectively, and CA in all types showed strong variability (40.7% < CV < 92.3%). However, the value of LAI was close to 2.40 for all types, and had less variation than other morphological indicators, which showed weak or medium variation. Overall, it varied greatly for morphological characteristics among different types or different tree species in one plant configuration type by comprehensively considering DBH, H, CA and LAI.Average value of four morphological indicators of landscape tree species in six plant configuration types were displayed in Table CHI, Pn, Er, Ci and Gs showed different regularities among six plant configuration types or tree species for a same type . Besides, K. paniculata had the lowest wCO2 and WCO2, which only accounted for 54.2% and 34.8% of G. biloba, respectively, and S. babylonica had the lowest QCO2, which accounted for 28.6% of P. stenoptera . Also, the lowest wCO2, WCO2 and QCO2 were observed both in J. formosana . P. Cerasys had the lowest wCO2, while C. chinensis had the largest WCO2, which accounted for 42.4% and 38.5% of A. persista 'Duplex', respectively. Different from wCO2 and WCO2, the top three tree species of QCO2 were shown as C chinensis > P. Cerasys > P. cerasifera 'Atropurpure', and the largest QCO2 was observed in S. oblata, which only accounted for 28.4% of C. chinensis. From Fig. wCO2, WCO2 and QCO2 of O. fragrans were significantly higher than those of P. serratifolia and V. odoratissimum, but there was no significant difference between this two species (P < 0.05).Figure wCO2 and WCO2 were shown as A. triloba > P. granatum > C. praecox > L. quihoui, and A. triloba was 1.9 ~ 3.5 times and 2.2 ~ 5.2 times higher than other tree species, respectively. But QCO2 was shown as C. praecox > A. triloba > P. granatum > L. quihoui, and there was no significant difference between the four species (P < 0.05). From Fig. B. megistophylla had the largest wCO2, which was 1.0 times higher than P. glabratum and 1.7 times higher than B. sinica var. parvifolia. And P. glabratum had the largest WCO2 and QCO2, which was 1.8 times and 2.0 times higher than B. megistophylla, respectively, and 7.5 times and 8.7 times higher than B. sinica var. parvifolia, respectively.Figure wCO2, WCO2 and QCO2 of DLA were significantly higher than those of ELA, and wCO2, WCO2 and QCO2 of DSA were also significantly higher than those of ESA, but it had no significant difference between DS and ES (P < 0.05). On the whole, wCO2 and WCO2 were shown as SA > S > LA, while QCO2 was shown as LA > SA > S, and there was no obvious difference of wCO2, WCO2, QCO2 between SA and S (P < 0.05)  analysis method was used to analyze the correlation between photosynthetic and morphological characteristics and carbon sequestration potential  in LA, SA and S. Results displayed that for LA, SA and S, the characteristic values of RDA on the first ordination axis were 0.7734, 0.5648, 0.769, respectively, and were 0.1564, 0.3854 and 0.1845 on the second ordination axis, respectively. Besides, the cumulative interpretation rates of the first and second axes were 92.98%, 95.02%, 95.35% for LA, SA and S, respectively, and the overall interpretation rates were 98.78%, 98.75%, 98.92%, respectively (Table P < 0.05), which suggested a statistically significant result. Further analysis found that the first two ordination axes of RDA of three types can better reflect the correlation between various indicators of tree species and carbon sequestration potential, and their correlation was mainly determined by the first ordination axis.Redundancy , Pn , LAI , CHI and DBH  had a significant impact on trees comprehensive carbon sequestration potential, and the correlation was shown as CA > Pn > LAI > CHI > DBH, with corresponding explanatory amounts of 46.4%, 32.9%, 7.2%, 4.4% and 2.6%, respectively , Fn , LAI  had a significant impact on trees comprehensive carbon sequestration potential, and the correlation was shown as CA > Pn > LAI, with corresponding interpretation amounts of 45.6%, 41.4% and 7.3%, respectively , Pn , CA  and Gs had a significant impact on trees comprehensive carbon sequestration potential, and the correlation was shown as LAI > Pn > CA > Gs, with corresponding interpretation amounts of 38.9%, 29.1%, 25.2% and 1.7%, respectively , small arbors (SA), shrubs (S) accounted for 53%, 25%, 22% for,respectively, and deciduous trees (D), evergreen trees (E) accounted for 56%, 44%, respectively. It was basically consistent with the current vegetation type of local urban green space, and represented the requirements for plant selection in the area's landscape construction.CO2 has been gradually emphasized in recent years, and many relevant researches have been carried out [Pn for D was higher than that of E, which was consistent with the conclusion of Guo Hui's assessment on carbon fixation and oxygen release of 15 native common landscape tree species in Zhengzhou [wCO2 of S was larger in contrast with LA and it exhibited a similar characteristics of plants in some southern inland cities of China [LAI of different types in our study made a comparatively similar regularity between wCO2 and WCO2, while the large disparity of crown width made them an obvious discrepancy with QCO2, and caused QCO2 of LA and SA exceeded that of S. Therefore, arbor trees played an important ecological role in landscaping construction system. Shrubs had a strong carbon sequestration potential, but their small morphological characteristics lead to less total carbon sequestration. Therefore, LAI and crown width can be increased through dense planting methods such as group planting and cluster planting, so as to increase their total carbon fixation [The function of urban green space in mitigating ried out , 16, 54.hengzhou , but we hengzhou –61. Howehengzhou , 63, sinof China , 64, whiof China , 50. Besfixation .A. triloba(9.8 gm−2 d−1), A. persica ‘Duplex’(8.7 gm−2 d−1) and P. cerasifera 'Atropurpurea'(5.6 gm−2 d−1) had a higher wCO2, and P. stenoptera(4.0 gm−2 d−1), S. babylonica(3.8 gm−2 d−1), P. acerifolia(3.5 gm−2 d−1), C. camphora(2.2 gm−2 d−1) had a lower wCO2. However,,wCO2 of C. camphora in northern Zhejiang Province was large (11.374 gm−2 d−1) while P. cerasifera ‘Atropurpurea’ was small (2.178 gm−2 d−1) [wCO2 of A. triloba (6.79 gm−2 d−1) in Shenyang was smaller compared with other six common garden plants, such as Q. mongolica and P. alba × P.beriliensis [wCO2 of C. camphora (6-10 gm−2 d−1) in Shanghai was lower than that of S. babylonica (> 12 gm−2 d−1) [WCO2 of M. grandiflora was 6.1 g.m−2.d−1 in Zhengzhou, but it was 46.49 g.m−2.d−1 in Shanghai’s garden plants community [−2 d−1 in Kunming’s road greening trees [WCO2 seemed to present an increasing trend from north to south in China. In addition, several arbor trees, such as P. stenoptera, P. acerifolia and C. camphora, showed high in QCO2 (exceeding 400 g d−1) in our study, which was similar to the conclusions of Guo Hui and Zhao Yanling [Carbon sequestration potential also varied greatly among tree species in a same plant configuration type. For examples, our results showed that m−2 d−1) . Except iliensis . And wCOm−2 d−1) . We alsoommunity  and 174.ng trees . This su Yanling , 58.wCO2 reflected the strength of plants carbon sequestration potential through their leaf photosynthesis, and for that, it was directly applied for evaluating the carbon sequestration potential of different tree species [QCO2 could better represent the comprehensive level of carbon sequestration of individual plants and ecological function of urban green space, which also was considered a better scientific evaluation standard [QCO2 into account at the same time [wCO2, WCO2 and QCO2 respectively represented three different perspectives of plants carbon sequestration potential, and appropriate indicators should be selected under various conditions for analysis. Specifically, tree species with high wCO2 can be given priority on the condition of thinking about plants photosynthetic carbon fixation capacity, and tree species with high WCO2 could be considered in the case of limited urban green space, and tree species with high QCO2 should be suggested when the number of seedlings was determined. However, we preferred that tree species with both higher wCO2, WCO2 and QCO2 were recommended, such as Populus, P. stenoptera, P. acifolia in LA, P. serratifolia, O. fragrans, S. oblata in SA, and B. sinica var. parvifolia, B. megistophylla, L. quihoui in S. species , 66. Howstandard , 62, 65.ame time . Besidesame time , 68. NevPn and Er were extremely significant correlated with wCO2 [LAI and DBH were extremely significant correlated with WCO2, while CD, DBH, H and LAI were extremely significant correlated with QCO2, respectively [wCO2, WCO2 and QCO2, and found that CA, LAI and Pn could significantly affect the comprehensive carbon fixation potential of landscape tree species both in LA, SA and S.Previous studied showed that ith wCO2 , 69, andectively . This waBriefly, arbor trees should be chosen as the main greening plants, while shrubs and herbs as the auxiliary plants in terms of the allocation mode of urban greening vegetation, and higher carbon sequestration potential tree species in different plant configuration types should also be selected. Meanwhile, we’d better consider an appropriate proportion of deciduous and evergreen tree species to build a multi-layer community structure with high biodiversity, so that it can reasonably use water, light, temperature, space and other resources, to improve the ecological and economic comprehensive benefits of the entire urban green space. In this study, we didn’t access the carbon sequestration potential of herbs, so it will be considered in the future research. Moreover, the work we did lean towards a fundamental research, and our analysis about the assessment of carbon sequestration potential of trees was mainly based on trees photosynthetic capacity. However, trees carbon sequestration potential were also related to their biomass, age, growth environment conditions, management level and so on, so we will consider these factors comprehensively by combing practical applications and try to provide a more appropriate method for city policymakers. Besides that, we provided a ranking list of 32 common landscape trees based on the condition of our study areas, which may give a basis for the selection of landscape tree species in Zhengzhou or similar areas.wCO2, WCO2 and QCO2. Comprehensive evaluation results of wCO2, WCO2 and QCO2 by cluster analysis showed that the top 10 tree species of high carbon sequestration potential include 3 DLA (Deciduous large arbor), 2 ELA (Evergreen large arbor), 1 DSA , 2 ESA  and 2 ES (Evergreen shrubs). Considering that hierarchical landscape spaces are generally formed in urban landscaping, we recommend using Populus, P. stenoptera, P. acerifolia among LA (Large arbors), and P.serratifolia, O. fragrans, S. oblata among SA , and B. sinica var. parvifolia, B. megistophylla, L. quihoui among S (Shrub). Pn, CA and LAI were the main factors which affected the comprehensive carbon sequestration potential of different plant configuration types.It had a different carbon sequestration potential among the selected 32 common landscape tree species by comparing their