Substances In Serum Research Articles

Detection of TNF-α using the established ab-MPs-CLIA

Tumor necrosis factor alpha (TNF-α) is a key cytokine in inflammation and immune responses, making its rapid and accurate detection essential for disease diagnosis and management. In this study, we developed a highly sensitive chemiluminescence immunoassay (CLIA) using antibody-coated magnetic particles (Ab-MPs-CLIA) for TNF-α detection. From nine candidate antibodies, we identified an optimal pair through epitope competition and affinity assessments, significantly improving assay performance. The Ab-MPs-CLIA achieved a detection limit of 0.25 pg/mL, 6.8 times more sensitive than Siemens commercial kits, with a broad linear range of 9.2–1077 pg/mL. The method demonstrated excellent stability, both under accelerated conditions at 37 °C for 7 days and long-term storage at 4 °C for 12 months. It showed no cross-reactivity with common interfering substances in human serum, ensuring high specificity. Notably, the entire process, from sample preparation to result, takes just 25 min, compared to 3–4 h for both ELISA and RIA, and CLIA typically offers 10–100 times higher sensitivity than these methods. These advantages make the Ab-MPs-CLIA an ideal option for clinical laboratories, providing superior sensitivity, specificity, broader dynamic range, and greater operational efficiency than existing TNF-α detection technologies.

12239 Effect Of Nano2, A Donor Of Nitric Oxide, On Glucose Mediated Oxidative Stress In Pancreaticb-cells

Abstract Disclosure: K.N. Islam: None. R.K. Islam: None. M.B. Rashid: None. B. Smith: None. Diabetes is a leading cause of morbidity and mortality. Elevation of oxidative stress is a common cause in diabetic patients because of higher levels of blood sugar concentration. Nitric oxide (NO) is a gasotransmitter that has many health benefits for many disease conditions. NO also has an antioxidant property. The pancreatic β-cells are insulin-secreting cells. The present study was undertaken to investigate the effects of NaNO2, a donor of NO on glucose-induced oxidative stress in pancreatic β-cells (HIT) from a Chinese hamster. First, we determined the effects of dose responses of glucose (25, 50, 75, 100, and 150 mM) on the induction of oxidative stress as it was measured by the formation of thiobarbituric acid reactive substances (TBARS) in serum starved β-cells. We observed marked oxidative stress in the serum starved β-cells by the treatment with 75 mM of glucose. Later, we tested the effects of NaNO2 on glucose-induced oxidative stress in serum starved β-cells, and we found that NaNO2 markedly inhibited glucose-induced oxidative stress in serum starved β -cells. Treatment with NaNO2 caused significant induction of cell viability and a marked reduction of cellular cytotoxicity in β-cells under serum starvation incubated without or with glucose. In addition, NaNO2 treatment of β-cells antagonized expression levels of inflammatory cytokines, TNF-α and inhibited the activation of the transcription factor, NF-κB signaling pathway. Our findings suggest that NO or NO donors may be used as therapy to cure diabetes via inhibiting oxidative stress thereby decreasing blood sugar levels in diabetic patients. Presentation: 6/3/2024

RELATIONSHIP BETWEEN SERUM SUBSTANCE P LEVELS AND ACNE VULGARIS

This investigation aimed to see if serum substance P levels and acne vulgaris were related. This study applies the case-control method to an observational analytical investigation. Samples were collected at KSM Dermatology's Cosmetic Division Polyclinic. Blood samples were evaluated at the UI Faculty of Medicine, Integrated Laboratory, and venereology at UI Hospital, Medan. Each group in this study included a minimum of 18 samples. The study indicates a relationship between serum substance P levels and acne vulgaris. Acne vulgaris is 5.4 times more likely to occur in people with high serum substance P levels. greatest in males. The average serum substance P level in acne vulgaris is 62.6 ± 32.4 pg/ml, and the highest levels can be found in the age group of 28 to 32 years. The average serum substance P level of acne vulgaris based on gender is highest in men.

Transplacental transfer of perfluorinated and poly-fluorinated substances in maternal-cord serum and association with birth weight: A birth cohort study, China

Although the effects of traditional perfluorinated and polyfluorinated substances (PFASs) exposure have been extensively explored, research on novel PFASs remains limited, and there is a lack of data regarding their placental transfer and fetal impact. Herein, we aimed to examine maternal and fetal PFASs exposure levels, placental transfer efficiency (TTE), and the consequences of prenatal exposure on birth weight. The study included 214 mother-child pairs recruited in Wuxi birth cohort from 2019 to 2021. Twenty-three PFASs were quantified in maternal serum during the second trimester and umbilical serum during delivery. Median concentrations of ∑23PFASs in maternal and cord sera were 9.34 and 6.88 ng/mL, respectively. The novel alternatives exhibited elevated levels of maternal and fetal exposure, such as perfluorovaleric acid (PFPeA, 2.00 ng/mL and 1.66 ng/mL, respectively) and perfluorohexane sulfonate (PFHxS, 1.77 and 1.14 ng/mL, respectively). With increasing carbon chain length, the TTE of perfluorocarbonic acid (PFCAs) displayed a pattern of initially decreasing before subsequently increasing, with novel alternatives exhibiting a relatively high TTE. Multiple linear regression showed that exposure to perfluorobutane sulfonate (PFBS) and PFPeA in cord serum positively correlated with the birth weight of female infants (β = 231.04 g, 95% confidence interval [CI]: 21.73–440.36; β = 121.26 g, 95% CI: 29.51–213.00). No nonlinear relationship was observed between cord serum PFASs and birth weight. The weighted quantile sum (WQS) regression analysis has reaffirmed that PFPeA and PFBS were predominant contributors to the positive correlation observed between the mixture of PFASs and birth weight. Our findings suggest that novel PFASs may exhibit a heightened susceptibility for transplacental transfer and that exposure to PFBS and PFPeA during pregnancy could be linked to increased birth weight.

Enhanced peroxidase‐like activity of MnFe2O4 nanoparticles on halloysite nanotubes for uric acid detection

AbstractNanozymes have significantly advanced sensing assays by replicating native enzyme functions. However, designing nanozymes with high catalytic activity and easy recyclability remains challenging. The study presented here has resulted in the development of a highly efficient and sensitive colorimetric system for the detection of uric acid, utilizing MnFe2O4@HNTs—a novel composite material consisting of MnFe2O4 loaded onto halloysite nanotubes. These nanocomposites exhibited outstanding peroxidase‐like activity and attractive magnetic properties. The catalytic efficiency of the MnFe2O4@HNTs in the oxidation of 3,3′,5,5′‐tetramethylbenzidine, in the presence of H2O2, was remarkable, leading to a distinct color change from colorless to blue. A linear relationship was observed between absorbance and UA concentration in the range of 1–20 μM, with a detection limit as low as 52 nM. Mechanistic investigations revealed that reactive oxygen species (ROS), specifically singlet oxygen (1O2), were generated through the decomposition of H2O2, which is responsible for the peroxidase‐like activity demonstrated by the MnFe2O4@HNTs. The method showed minimal interference from serum substances and high selectivity. Magnetic MnFe2O4 allowed easy separation and maintained over 95% activity after seven reuse cycles. The developed assay was successfully applied to the detection of uric acid in human serum, achieving recoveries greater than 98.60%. This research significantly advances the design of recyclable high‐performance nanozymes and establishes an effective colorimetric sensing platform for UA detection in clinical samples, potentially improving diagnostic tools for healthcare applications.

264. SIGNIFICANCE AND USEFULNESS OF PLASMA SUBSTANCE P CONCENTRATION IN PREDICTING HIGH-RISK PATIENTS FOR ASPIRATION AFTER ESOPHAGEAL CANCER SURGERY

Abstract Background The greatest challenge in postoperative complications of esophageal cancer is the prevention of pulmonary complications, especially aspiration pneumonia. Although the introduction of minimally invasive surgery and intervention in breathing and swallowing rehabilitation can improve the motor function of breathing and swallowing, we reported the possibility that a decline in the swallowing reflex may play a significant role in esophageal cancer patients, many of whom are elderly. In fact, about 80% of esophageal cancer patients in Japan are over 60 years old, and 53% of them have asymptomatic cerebral infarction, about half of which are reported to be found near the basal ganglia, which control the swallowing reflex. The swallowing reflex is controlled by the secretion of substance P, synthesized in the cervical sympathetic ganglia, into the pharynx and trachea upon stimulation of dopamine synthesized in the basal ganglia. Methods To test the hypothesis that patients with preoperative subclinical cerebral infarction are more likely to develop aspiration postoperatively due to decreased substance P secretion. To compare the association between the presence or absence of cerebral infarction near the basal ganglia and serum substance P levels and the development of postoperative aspiration or aspiration pneumonia due to videoendoscopy and video fluorography in patients with thoracic esophageal cancer scheduled for video-assisted esophagectomy by preoperative brain MRI imaging and blood sampling. Results We report the results of an interim analysis of 54 cases in which the measurement of substance P was completed. Male: 39 cases, female: 15 cases. Mean age: 69.3 (49-82) years. Tumor location: Ut/Mt/Lt=13/25/14. MRI showed infarction in or near the basal ganglia in only 2 cases. Median preoperative blood substance P concentration: 48.2 pg/ml. Eleven one patients had postoperative aspiration. Preoperative substance P levels ≥48.2 pg/ml were defined as the High group, and those below 48.2 pg/ml were defined as the Low group. Postoperative aspiration was observed in 2/27 (7.4%) patients in the High group, while 9/26 (34.6%) patients in the Low group, which was significantly higher (p=0.015). Conclusion In fact, subclinical cerebral infarction was much less common than reported and was not useful as a predictor of postoperative aspiration. On the other hand, patients with low preoperative blood substance P levels had a very high risk of developing aspiration postoperatively due to impaired swallowing reflexes, suggesting that preoperative blood substance P levels may be useful in assessing postoperative aspiration risk in patients with thoracic esophageal cancer.

A randomized controlled trial of a housing intervention to reduce endocrine disrupting chemical exposures in children

Few studies have considered household interventions for reducing endocrine disrupting chemical (EDC) exposures. We conducted a secondary analysis of a randomized controlled trial, originally designed to reduce lead exposure, to evaluate if the intervention lowered EDC exposures in young children. Study participants were children from the Cincinnati, Ohio area (n = 250, HOME Study). Prenatally, families received a housing intervention that included paint stabilization and dust mitigation, or as a control, injury prevention measures. At 24-months, we measured organophosphate esters (OPEs) and phthalates or their metabolites in dust and urine. We measured perfluoroalkyl substances (PFAS) in dust and serum at 24- and 36-months, respectively. We assessed associations between dust and biomarker EDCs using Spearman correlations, characterized EDC mixtures via principal components analysis, and investigated treatment effects using linear regression. To mitigate selection bias, we fit statistical models using inverse probability of retention weights. Correlations between dust EDCs and analogous biomarkers were weak-to-moderate (ρ’s ≤ 0.3). The intervention was associated with 23 % (95 % CI: −38, −3) lower urinary DEHP metabolites and, in a per-protocol analysis, 34 % lower (95 % CI: −55, −2) urinary MBZP. Additionally, among Black or African American children, the intervention was associated with lower serum concentrations of several PFAS (e.g., −42 %; 95 % CI: −63, −8 for PFNA). Household interventions that include paint stabilization and dust mitigation may reduce childhood exposures to some phthalates and PFAS in Blacks/African Americans. These findings highlight the need for larger studies with tailored and sustained housing interventions.

The toxicity of cisplatin derives from effects on renal organic ion transporters expression and serum endogenous substance levels

Acute kidney injury (AKI) is a worldwide public health problem with high morbidity and mortality. Cisplatin is a widely used chemotherapeutic agent for treating solid tumors, but the induction of AKI restricts its clinical application. In this study, the effect of cisplatin on the expression of organic ion transporters was investigated through in vivo and in vitro experiments. Targeted metabolomics techniques were used to measure the levels of selected endogenous substances in serum. Transmission electron microscopy was used to observe the microstructure of renal tubular epithelial cells. Our results show that the toxicity of cisplatin on HK-2 cells or HEK-293 cells was time- and dose-dependent. Administration of cisplatin decreased the expression of OAT1/3 and OCT2 and increased the expression of MRP2/4. Mitochondrial damage induced by cisplatin lead to renal tubular epithelial cell injury. In addition, administration of cisplatin resulted in significant changes in endogenous substance levels in serum, including amino acids, carnitine, and fatty acids. These serum amino acids and metabolites (α-aminobutyric acid, proline, and alanine), carnitines (tradecanoylcarnitine, hexanylcarnitine, octanoylcarnitine, 2-methylbutyroylcarnitine, palmitoylcarnitine, and linoleylcarnitine) and fatty acids (9E-tetradecenoic acid) represent endogenous substances with diagnostic potential for cisplatin-induced AKI.

Effect of classical music on light-plane anaesthesia and analgesia in dogs subjected to surgical nociceptive stimuli

The objectives of this prospective, randomized, blinded, crossover, experimental study were to detect the potential anaesthetic- and analgesic-sparing effects of classical music provided to dogs undergoing skin surgery, and to investigate the role of substance P as an intraoperative pain indicator. Twenty dogs were included, each subjected to three different treatments: Chopin music, Mozart music and no music. They were premedicated with acepromazine, butorphanol and meloxicam and anaesthetized with propofol and isoflurane. Fentanyl was used as rescue analgesia. The anaesthetic depth was monitored by using the bispectral index along with standard anaesthetic monitoring, and autonomic nervous system responses were used to monitor the adequacy of analgesia. Furthermore, measurements of substance P serum concentration were carried out. Dogs exposed to music required less isoflurane and fentanyl. Furthermore, a statistically significant effect of time on substance P concentration was observed regardless of exposure to music, and there was a significant interaction effect between different timepoints and the type of acoustic stimulus. Classical music seems to have an isoflurane and fentanyl sparing effect on dogs undergoing minor surgery. Following surgical stimulation, the serum substance P concentration increases rapidly, and thus appears to be a potentially useful pain indicator.

Perinatal Exposure to Phenols and Poly- and Perfluoroalkyl Substances and Gut Microbiota in One-Year-Old Children.

The role of the gut microbiota in human health calls for a better understanding of its determinants. In particular, the possible effects of chemicals with widespread exposure other than pharmaceuticals are little known. Our aim was to characterize the sensitivity of the early-life gut microbiota to specific chemicals with possible antimicrobial action. Within the SEPAGES French couple-child cohort, we assessed 12 phenols in repeated urine samples from 356 pregnant women and their offspring and 19 poly- and perfluoroalkyl substances (PFASs) in serum from the pregnant women. We collected stool samples from the children at one year of age, in which the V3-V4 region of the 16S rRNA gene was sequenced, allowing for gut bacterial profiling. Associations of each chemical with α- and β-diversity indices of the gut microbiota and with the relative abundance of the most abundant taxa were assessed using single-pollutant and mixture (BKMR) models. Perinatal exposure to certain parabens was associated with gut microbiota α- and β-diversity and with Firmicutes and Proteobacteria. Suggestive associations of certain phenols with genera of the Lachnospiraceae and Enterobacteriaceae families were observed, but these were not maintained after correction for multiple testing. Parabens, which have known antimicrobial properties, might disrupt the child gut microbiota, but larger studies are required to confirm these findings.

Targeted quantification of per- and polyfluoroalkyl substances (PFASs) in livestock serum by liquid chromatography-high-resolution mass spectrometry.

Per- and polyfluoroalkyl substances (PFASs) have attracted increasing attention due to their persistence in the environment and potential adverse effects on human and animal health. The detection and quantification of PFASs in livestock could substantially contribute to monitoring their presence within the food chain. We developed a targeted quantification method for 34 PFASs in livestock serum by liquid chromatography-high-resolution mass spectrometry (LC-HRMS). We used protein precipitation for serum sample extraction and accurate mass measurement of targeted PFAS compounds for quantification. We validated the method with various analytical parameters, achieving accuracy of 70-120% and precision of <20%. The method also demonstrated good analytical sensitivity, with a limit of detection of <0.051 ng/mL and a limit of quantification of <0.175 ng/mL. When applying the developed method to actual serum samples from a variety of livestock, we successfully identified and quantified various PFASs in different livestock species. Our method has the potential to be a valuable tool for veterinary laboratory analysis of PFAS contamination in livestock.

Per- and polyfluoroalkyl substances in serum and breastmilk samples among pregnant farmworkers in Thailand

Per- and polyfluoroalkyl substances in serum and breastmilk samples among pregnant farmworkers in Thailand

Quantifying levels of per- and polyfluoroalkyl Substances (PFAS) in water and serum after contamination from agricultural biosolid application

The National Academies of Sciences, Engineering, and Medicine recommends per- and polyfluoroalkyl substance (PFAS) blood testing for patients with risk of elevated exposure, and the Agency for Toxic Substances and Disease Registry (ATSDR) suggests PFAS blood testing based on exposure. Barriers to PFAS blood testing include cost, access to labs, and evolving laboratory methods. We quantify water and serum PFAS levels among a highly-exposed cohort in an area with groundwater contaminated by historical agricultural biosolid application. We compare the gold standard PFAS serum test with a commercial test and results from a one-compartment toxicokinetic model. Participants were adults (n = 30) whose household (n = 19) water had levels of the sum of six PFAS > 500 ng/L. Serum PFAS were measured using liquid chromatography-tandem mass spectrometry. Demographic and water consumption data were collected via telephone. Serum PFAS results from the commercial test were accessed via medical record. Statistical analysis included descriptive statistics and bivariate plots of serum levels. Perfluorohexanoic acid, perfluoroheptanoic acid (PFHpA), perfluorooctanoic acid (PFOA), perfluorononanoic acid (PFNA), perfluorobutanesulfonic acid, perfluorohexanesulfonic acid (PFHxS), and perfluorooctanesulfonic acid (PFOS) were detected in 19 wells, and PFHpA, PFOA, PFNA, perfluorodecanoic acid, perfluoroundecanoic acid, PFHxS, and PFOS were detected in at least 19 participants’ serum. In well water, PFOA and PFOS levels had geometric means (GMs) of 1749 ng/L (geometric standard deviation [GSD] 2.4) and 887 ng/L (GSD 19.7), respectively. In serum, PFOA and PFOS had GMs of 116.2 µg/L (GSD 13.5) and 58.3 µg/L (GSD 13.8), respectively. Our results are comparable with and had a wider mix of PFAS than other high-exposure cohorts. There was good agreement between the commercial and gold standard tests for PFOA, PFNA, and PFHxS, and mixed agreement between the gold standard test and modeled predictions, suggesting water-based toxicokinetic models of serum PFAS may be inadequate for assessing exposure in this population.

Associations of perfluoroalkyl substances with metabolic-associated fatty liver disease and non-alcoholic fatty liver disease: NHANES 2017-2018.

This study investigated the potential effects of perfluoroalkyl substance (PFAS) in serum on MAFLD, NAFLD, and liver fibrosis. Our sample included 696 participants (≥ 18years) from the 2017-2018 NHANES study with available serum PFASs, covariates, and outcomes. Using the first quartile of PFAS as the reference group, we used weighted binary logistic regression and multiple ordered logistic regression used to analyze the relationship between PFAS and MAFLD, NAFLD, and liver fibrosis and multiple ordinal logistic regression to investigate the relationship between PFAS and MAFLD, NAFLD, and liver fibrosis and calculated the odds ratio (OR) and 95% confidence interval for each chemical. Finally, stratified analysis and sensitivity analysis were performed according to gender, age, BMI, and serum cotinine concentration. A total of 696 study subjects were included, including 212 NAFLD patients (weighted 27.03%) and 253 MAFLD patients (weighted 32.65%). The quartile 2 of serum PFOA was positively correlated with MAFLD and NAFLD (MAFLD, OR 2.29, 95% CI 1.05-4.98; NAFLD, OR 2.37, 95% CI 1.03-5.47). PFAS were not significantly associated with liver fibrosis after adjusting for potential confounders in MAFLD and NAFLD. Stratified analysis showed that PFOA was strongly associated with MAFLD, NAFLD, and liver fibrosis in males and obese subjects. In women over 60years old, PFHxS was also correlated with MAFLD, NAFLD, and liver fibrosis. The serum PFOA was positively associated with MAFLD and NAFLD in US adults. After stratified analysis, the serum PFHxS was correlated with MFALD, NAFLD, and liver fibrosis.

A green analytical method for the simultaneous determination of 17 perfluoroalkyl substances (PFAS) in human serum and semen by ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS)

The ubiquity of perfluoroalkyl substances has raised concerns about the unintended consequences of PFAS exposure on human health. In the present study, an eco-friendly ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method was developed for the simultaneous determination of 17 PFAS in human serum and semen samples. QuEChERS salts MgSO4:NaCl 4:1 (w/w) were used for the extraction. The separation of analytes was performed on an ACQUITY BEH C18 column (100 × 2.1 mm, 1.7 μm), using water:methanol 95:5 and methanol as mobile phases A and B, respectively, both containing 2 mM ammonium acetate. Multiple reaction monitoring (MRM) in negative ion mode was used, selecting two transitions for each analyte, except for perfluorobutanoic acid (PFBA) and perfluoropentanoic acid (PFPeA). The analytical method was validated according to the Organization of Scientific Area Committees (OSAC) for Forensic Sciences guidelines and AGREE approach software was used to evaluate the greenness of the method. The developed procedure was applied to the analysis of 10 paired human serum and semen samples, proving the suitability in high throughput laboratories due to the easy preparation and the reduced volume of toxic solvents. Moreover, it allows to perform further investigation on the correlation between serum and semen PFAS concentration, focusing on male reproductive system correlated pathologies, such as male infertility.

Integration of ultrashort-chain compounds into the biomonitoring of per- and polyfluorinated substances in human plasma and serum

Ultrashort-chain (USC) per- and polyfluoroalkyl substances (PFAS) are small and very polar compounds with carbon chain lengths of shorter than C4. Their ubiquitous and high levels of occurrence in environmental aquatic systems have raised significant concern in conjunction with long-chain PFAS contamination. Measuring USC PFAS in blood can not only monitor human exposure but also serves as a valuable tool for studying the potential risks associated with USC PFAS exposure. The high polarity of USC PFAS poses a challenge to current analytical practices based on the reversed-phase liquid chromatography, primarily due to insufficient chromatographic retention. In this study, a simple and reliable workflow was developed for the simultaneous analysis of C1 to C10 perfluoroalkyl carboxylic and sulfonic acids, along with four alternative PFAS, in human plasma and serum. The chromatographic analysis was conducted using a polar-embedded reversed-phase LC column. Fetal bovine serum (FBS) was chosen for method validation due to its absence of all analytes, except for trifluoroacetic acid (TFA). Subsequently, a TFA isotope, 13C-TFA, was employed as a surrogate to assess the method accuracy for TFA in FBS. A single-step sample preparation procedure, conducted by mixing 100 µL of FBS with 200 µL of methanol, was demonstrated to be effective for the accurate and precise analysis of fortified FBS samples. Ten isotopes of C3 to C10 PFAS, serving as extracted internal standards, were added to the samples at 1 ppb to validate the accuracy of the entire workflow. Calibration standards were prepared in reverse osmosis water due to its cleanliness for all analytes. Phosphate-buffered saline was incorporated into the calibration standard solution to achieve similar chromatographic performance between standard and sample solutions. The calibration ranges varied among different analytes, spanning from 0.05 – 40 ppb, 0.1 – 40 ppb, 0.25 – 40 ppb, and 0.5 – 40 ppb. Method accuracy and precision were evaluated at three fortification levels, ranging from 0.4 to 30 ppb. All analytes and extracted internal standards exhibited recovery values within 20 % of the nominal concentration across all fortification levels. Satisfactory method precision was demonstrated with%RSD values <12 %. The validated method was applied to measure PFAS in NIST 1950 and 1957 standard reference human plasma and serum, affirming the established workflow is suitable for accurate quantification of C1 to C10 PFAS in human plasma and serum matrices.

Changes in the Serum and Tissue Levels of Free and Conjugated Sialic Acids, Neu5Ac, Neu5Gc, and KDN in Mice after the Oral Administration of Edible Bird’s Nests: An LC–MS/MS Quantitative Analysis

Edible bird’s nests have a variety of biological activities, the main components of which are sialic acids. Sialic acids are a group of nine-carbon N-acetylated derivatives of neuraminic acid containing a keto group at position C2 and play important roles in many biological processes. To verify whether the oral administration of edible bird’s nests would change the content and distribution of sialic acid components in vivo, a liquid chromatography–mass spectrometry method for the quantitative analysis of sialic acid levels in serum and tissues was developed. In the negative ion mode, the mobile phases consist of 0.1% formic acid in water (A) and acetonitrile (v/v) (B). Isocratic elution was performed with 60% B for 0−15 min. The chromatographic separation was performed on a Morphling HILIC Amide column (2.1 mm × 150 mm, 5 μm) at a flow rate of 0.5 mL min−1. The results showed that the correlation coefficients of the typical calibration curves were all higher than 0.995, exhibiting good linearity. The levels of free and conjugated forms of N-glycolylneuraminic acid (Neu5Gc), N-acetylneuraminic acid (Neu5Ac), and 2-keto-3-deoxy-D-glycero-D-galactonononic acid (KDN) in the serum and different tissues were simultaneously detected after the oral administration of the edible bird’s nests at a daily dose of 300 and 700 mg Kg−1 for seven days in mice. Our study found that the oral administration of edible bird’s nests can significantly increase the concentration of total sialic acids (Neu5Gc + Neu5Ac + KDN) in serum and spleen and lungs tissues, which may be related to the anti-inflammatory and immune function of edible bird’s nest, but further studies are needed to verify this. Neu5Ac was the dominant sialic acid in brain tissue, and Neu5Gc was the dominant sialic acid in serum and other tissues, including heart, liver, spleen, lungs, and kidney. Moreover, we found that the forms of Neu5Ac and Neu5Gc were mainly conjugated in all groups except liver tissue. In conclusion, the method we established had good linearity and accuracy; it allowed the analytes to be effectively separated from the matrix and endogenous substances in serum or tissues, so it could effectively detect the distribution and concentration of free and conjugated forms of sialic acids in serum and tissues, which was beneficial to the research and exploitation of edible bird’s nests and sialic acids.

Role of substance P in cerebral edema and association with an estimated specific gravity of the brain and an outcome prediction in post-traumatic cerebral edema

PurposeThe study aims to evaluate the role of substance P in cerebral edema and outcomes associated with acute TBI. MethodPatients with acute TBI who presented within 6 h and a CT scan showed predominantly cerebral edema were included in the study. Substance P level was assessed from a serum sample collected within 6 h of trauma. We also evaluated the brain-specific gravity using the Brain View software. ResultA total of 160 (128 male) patients were recruited. The median serum substance P concentration was 167.89 (IQR: 101.09–238.2). Substance P concentration was high in the early hours after trauma (p = 0.001). The median specific gravity of the entire brain was 1.04. Patients with a low Glasgow coma scale (GCS) at admission had a high concentration of the substance P. In the univariate analysis, low GCS, elevated serum concentrations of substance P level, high Rotterdam grade, high cerebral edema grade, a high international normalized ratio value, and high blood sugar levels were associated with poor outcomes at six months. In logistic regression analysis, low GCS at admission, high cerebral edema grade, and elevated blood sugar level were strongly associated with poor outcomes at six months. The area under the receiver operating characteristic curve was 0.884 (0.826–0.941). ConclusionSerum substance P is strongly associated with the severity of cerebral edema after TBI. However, brain-specific gravity does not directly correlate with posttraumatic cerebral edema severity. Serum substance P does not influence the clinical outcome of traumatic brain injury.

A Phase I/IIa Prospective, Randomized, Open-Label Study on the Safety and Efficacy of Nebulized Liposomal Amphotericin for Invasive Pulmonary Aspergillosis.

Although nebulized liposomal amphotericin B (NLAB) is being used in invasive pulmonary aspergillosis (IPA) prophylaxis, no clinical trial has shown its efficacy as a therapeutic strategy. NAIFI is the inaugural randomized, controlled clinical trial designed to examine the safety and effectiveness of NLAB (dosage: 25 mg in 6 mL, three times per week for 6 weeks) against a placebo, in the auxiliary treatment of IPA. Throughout the three-year clinical trial, thirteen patients (six NLAB, seven placebo) were included, with 61% being onco-hematological with less than 100 neutrophils/μL. There were no significant differences noted in their pre- and post-nebulization results of forced vital capacity (FVC), forced expiratory volume in 1 s (FEV1), and oxygen saturation between the groups. Neither bronchospasm nor serum amphotericin B levels were reported in any patients given NLAB. 18F-Fluorodeoxyglucose positron emission tomography (FDG-PET-TC) was carried out at the baseline and after 6 weeks. A notable decrease in median SUV (standardized uptake value) was observed in NLAB patients after 6 weeks (-3.6 vs. -0.95, p: 0.039, one tail). Furthermore, a reduction in serum substance galactomannan and beta-D-Glucan was identified within NLAB recipients. NLAB is well tolerated and safe for patients with IPA. Encouraging indirect efficacy data have been derived from image monitoring or biomarkers. However, further studies involving more patients are necessary.

Molecular mechanism of residual lumbago and leg pain after transforaminal endoscopic treatment of lumbar disc herniation

To observe the residual of lumbago and leg pain with contained type （CT） and non-contained type （NCT） lumbar disc herniation （LDH） after transforaminal endoscopic treatment, and to explore the role of hypoxia-inducible factor-1α（HIF-1α） and transient receptor potential vanillate 1（TRPV1） pathway. A total of 68 single-segment LDH patients were selected from July 2021 to October 2022, including 44 males and 24 females；aged 26 to 67 years old with an average of（43.63±11.94） years old；course of disease was 4 to 36 （18.91±10.34） months；body mass index was （24.45±4.00） kg·m-2；there were 7 cases of L3,4 segments, 32 cases of L4,5 segments, and 29 cases of L5S1 segments. All of them were performed with percutaneous intervertebral endoscopic extraction of nucleus pulposus and were divided into contained group（CT group） and non-contained group （NCT group） with 34 cases respectively according to the integrity of outer layer of fibrous annulus observed during operation. A total of 17 patients who underwent open surgery for scoliosis or vertebral fracture were selected as control group, including 12 males and 5 females；aged 21 to 65 years old with an average of （39.41±12.80） years old；body mass index was （24.86±4.11） kg·m-2. The relative mRNA expression quantity of HIF-1α, TRPV1 in nucleus pulposus were measured by quantitative real-time PCR. The contents of neurokinin 1 receptor （NK1R）, nerve growth factor （NGF）, vascular endothelial growth factor （VEGF） in nucleus pulposus and the serum substance P （SP） and calcitonin gene-related peptide （CGRP） were detected by enzyme linked immunosorbent assay （ELISA）. The threshold of lumbar tenderness was detected by a pressure pain meter. The degree of lumbago and lumbar function were evaluated by visual analog scale （VAS） and Oswestry disability index （ODI） separately. The residual rate of postoperative lumbago and leg pain was assessed. The mRNA relative expression quantity of HIF-1α and TRPV1, and the contents of NK1R, NGF and VEGF in nucleus pulposus, and the levels of serum SP and CGRP before surgery in the NCT group were higher than those in the CT group（P<0.05）, and those in the CT group were higher than the control group（P<0.05）. At day 7 after surgery, the serum SP and CGRP levels, lumbago and leg pain VAS scores and lumbar ODI index in two LDH groups were lower than before surgery （P<0.05）, and those in the NCT group were higher than the CT group（P<0.05）, and the threshold of lumbar tenderness in the NCT group was lower than the CT group（P<0.05）. The differences of lumbago and leg pain VAS scores, lumbar ODI index and lumbar tenderness threshold between preoperative and postoperative 7 days in the NCT group were lower than those in the CT group（P<0.05）. The residual rate of lumbago and leg pain at 7 days after surgery in the NCT group was higher than that in the CT group（P<0.05）. HIF-1α and TRPV1 pathway promoted the excessive production of NGF, VEGF, NK1R in nucleus pulposus and serum neuropeptides SP and CGRP, which may lead to the higher residual rate of lumbago and leg pain with non-contained lumbar disc herniation postoperative.