Evaluation of protected benzoic acid on growth performance, nutrient digestibility, and gut health indices in starter pigs.

Benzoic acid is a common alternative for antibiotic and zinc oxide use in nursery diets. Free benzoic acid (BZA) is often supplied, but this form is absorbed before it can exert any effect on distal segments of the gut. The study aimed to evaluate the effects of protected benzoic acid on growth performance, nutrient digestibility, plasma metabolites, and gut health indices in starter pigs. A total of 192 pigs were weaned at 28 ± 1 d age (initial body weight, 8.72 ± 1.13 kg). Pens were assigned to one of four treatment diets (n = 8 pens per treatment): (1) no additive (NC), (2) free benzoic acid (BZA; 0.6%), (3) protected benzoic acid (BC50; 0.2%, supplied at a ratio of one to three equivalents of BZA), and (4) antibiotic growth promoter (AGP; Carbadox, 50 ppm). Diets were fed for three weeks over two periods (period 1, 7 d; period 2, 14 d). Body weight and feed intake were measured for each period. Feces were collected at the end of each period to determine apparent total tract digestibility (ATTD) of organic matter (OM), gross energy (GE), and crude protein (CP). One pig per pen was euthanized per period to determine plasma metabolites; jejunum and ileum morphology; jejunum, ileum, and colon cytokine abundance; and jejunum, ileum, and colon tight junction protein expression. The AGP group had increased average daily gain (ADG) and average daily feed intake (ADFI) compared to other groups in period 1 and overall (P < 0.05); however, ADG and ADFI of the BC50 group was intermediate between the NC and BZA groups and the AGP group in period 2. The ATTD of OM, GE, and CP were greater in the AGP group compared to the NC and BC50 groups (P < 0.05), whereas the BZA group was intermediate. Jejunum and ileum villus height and crypt depth increased from period 1 to period 2 (P < 0.01) but were similar across groups. Ileum and colon tumor necrosis factor-α (TNF-α) abundances were greater, whereas colon interleukin (IL)-1ß and colon and ileum IL-8 abundances were less, in the AGP group compared to the BZA group (P < 0.05); the NC and BC50 groups exhibited intermediate TNF-α, IL-1ß, and IL-8 abundance in the ileum and colon. Jejunum cytokine abundance did not vary among groups but declined from period 1 to period 2 (P < 0.05). Tight junction protein expression also did not vary among groups. In summary, protected BZA supported a slight increase in growth performance in starter pigs, suggesting its potential as an alternative feed additive in nursery diets.

Serum Metabolomic Profiles for Distinguishing Lung Cancer From Pulmonary Tuberculosis: Identification of Rapid and Noninvasive Biomarker.

BACKGROUND: Pulmonary tuberculosis (PTB) and lung cancer (LC) have similar clinical symptoms and atypical imaging findings, which are easily misdiagnosed. There is an urgent need for a noninvasive and accurate biomarker to distinguish LC from PTB. METHODS: A total of 694 subjects were enrolled and divided into discovery set (n = 122), identification set (n = 214), and validation set (n = 358). Metabolites were identified by multivariate and univariate analyses. Receiver operating characteristic curve were used to evaluate the diagnostic efficacy of biomarkers. RESULTS: Seven metabolites were identified and validated. Phenylalanylphenylalanine for distinguishing LC from PTB yielded an area under the curve of 0.89, sensitivity of 71%, and specificity of 92%. It also showed good diagnostic abilities in discovery set and identification set. Compared with that in healthy volunteers (median [interquartile range], 1.57 [1.01, 2.34] µg/mL), it was elevated in LC (4.76 [2.74, 7.08] µg/mL; ratio of median, [ROM] = 3.03, P < .01) and reduced in PTB (1.06 [0.51, 2.09] µg/mL; ROM = 0.68, P < .05). CONCLUSIONS: The metabolomic profile of LC and PTB was described and a key biomarker identified. We produced a rapid and noninvasive method to supplement existing clinical diagnostic examinations for distinguishing LC from PTB.

Increased PPARD Expression May Play a Protective Role in Human Lung Adenocarcinoma and Squamous Cell Carcinoma.

Peroxisome proliferator-activated receptor-δ, encoded by gene PPARD, is overexpressed in a majority of human lung cancer subtypes, but its role in the tumor progression remains poorly understood. We have analyzed the expression of PPARD in lung adenocarcinoma (LA) and squamous cell carcinoma (LSCC) datasets. The potential roles of PPARD in the pathological development of LA and LSCC were explored through literature-based pathway analysis and pathway enrichment analysis. In all LA datasets (N = 11) and in seven out of nine LSCC studies, the levels of PPARD were increased as compared to control tissues (log-fold changes were 0.37 ± 0.20 and 0.10 ± 0.37 for LA and LSCC, respectively). On average, the expression levels of PPARD in LA were higher than those in LSCC (p = 0.036). Pathway analysis showed that the overexpression of PPARD might play both positive and negative roles in the development of both LA and LSCC. Specifically, PPARD inhibits seven LSCC promoters and seven LA promoters and activates one LSCC inhibitor and another LA inhibitor. However, PPARD also activates six and one promoters of LA and LSCC, respectively, which would facilitate the development of LA/LSCC. Our results suggested a mixed role of PPARD in LA/LSCC, which may add new insights into the understanding of the PPARD-lung cancer relationship.

Plasma proteomic and metabolomic characterization of COVID-19 survivors 6 months after discharge.

Coronavirus disease 2019 (COVID-19) has gained prominence as a global pandemic. Studies have suggested that systemic alterations persist in a considerable proportion of COVID-19 patients after hospital discharge. We used proteomic and metabolomic approaches to analyze plasma samples obtained from 30 healthy subjects and 54 COVID-19 survivors 6 months after discharge from the hospital, including 30 non-severe and 24 severe patients. Through this analysis, we identified 1019 proteins and 1091 metabolites. The differentially expressed proteins and metabolites were then subjected to Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis. Among the patients evaluated, 41% of COVID-19 survivors reported at least one clinical symptom and 26.5% showed lung imaging abnormalities at 6 months after discharge. Plasma proteomics and metabolomics analysis showed that COVID-19 survivors differed from healthy control subjects in terms of the extracellular matrix, immune response, and hemostasis pathways. COVID-19 survivors also exhibited abnormal lipid metabolism, disordered immune response, and changes in pulmonary fibrosis-related proteins. COVID-19 survivors show persistent proteomic and metabolomic abnormalities 6 months after discharge from the hospital. Hence, the recovery period for COVID-19 survivors may be longer.

Serum levels of laminin and von Willebrand factor in COVID-19 survivors 6 months after discharge.

OBJECTIVES: The aim of this study was to evaluate the clinical characteristics, pulmonary diffusion function, chest computed tomography (CT), and serum lung cell damage indicators of coronavirus disease 2019 (COVID-19) survivors 6 months after discharge. METHODS: Data of COVID-19 survivors discharged from hospital between January 21, 2020 and January 11, 2021 and healthy controls were collected. Serum levels of surfactant protein D (SP-D)1, the receptor for advanced glycation end products (RAGE)2, laminin, and von Willebrand factor (vWF) were measured in the healthy controls and COVID-19 survivors 6 months after discharge. The relationships between serum lung cell damage indicator levels and various parameters were explored. RESULTS: Fifty-two COVID-19 survivors (31 with non-severe disease and 21 with severe disease) and 30 controls were included. Serum levels of laminin in COVID-19 survivors 6 months after discharge were significantly higher than those in the controls. The increase was more significant in elderly and female patients. Serum levels of RAGE and vWF were not statistically different from those of the controls. However, 6 months after discharge, COVID-19 survivors with abnormal chest CT and those in the severe group had higher vWF levels. CONCLUSIONS: COVID-19 patients had abnormal lung injury indicators 6 months after discharge. The recovery time after infection is currently unknown, and long-term observation is required.

A large-scale, multicentered trial evaluating the sensitivity and specificity of digital PCR versus ARMS-PCR for detecting ctDNA-based EGFR p.T790M in non-small-cell lung cancer patients.

BACKGROUND: Developing liquid biopsy technology with higher sensitivity and specificity especially for low-frequency mutations remains crucial. This study demonstrated superior performance of the newly developed digital PCR (dPCR) kit for ctDNA-based EGFR p.T790M detection in metastatic non-small-cell lung cancer (NSCLC) against ARMS-PCR. METHODS: This large-scale, multi-centered diagnostic study recruited 1,045 patients including 1,029 patients diagnosed with advanced NSCLC and 16 patients with specific samples between April 1st 2018 and November 30th 2019. EGFR p.T790M in plasma samples from mNSCLC patients were tested using dPCR with ADx-ARMS PCR and Cobas® EGFR Mutation Test V2 as comparator assays to confirm cut-off value for dPCR and evaluate its performance against ARMS-PCR-based assays. Efficacy was evaluated for patients with EGFR p.T790M detected by dPCR or ARMS-PCR, who underwent Osimertinib treatment. RESULTS: The sensitivity, specificity, and concordance of dPCR against ADx-ARMS PCR was 98.15%, 88.66% and 90.16%, respectively for 1,026 plasma samples. Additional 9.26% patients were detected positive by dPCR. The majority of those samples had a mutation allele frequency between 0.1% and 1%. In 45 paired tissue and plasma samples, the sensitivity improved from 30.77% to 53.85% by dPCR with the specificity over 90%. The response of Osimertinib in 74 EGFR p.T790M-positive patients detected by dPCR, including 26 determined as negative by ARMS-PCR, were evaluated to have an ORR of 44.59% and a DCR of 90.54%. CONCLUSIONS: dPCR is a sensitive and accurate tool for ctDNA-based EGFR p.T790M detection due to its significantly improved sensitivity without compromising specificity, and dPCR is equivalent to ARMS-PCR as a companion diagnostic tool while benefiting more patients under Osimertinib treatment. TRIAL REGISTRATION: Chinese Clinical Trial Registry identifier: ChiCTR2100043147.

Detection and predictors of anti-SARS-CoV-2 antibody levels in COVID-19 patients at 8 months after symptom onset.

Aim: To determine SARS-CoV-2 specific IgM and IgG levels of patients with COVID-19 at 8 months after symptom onset and to explore the predictors of antibody levels. Materials & methods: The magnetic chemiluminescence method was used to measure the antibody levels. Clinical data were collected and analyzed retrospectively. Results: A total of 54 patients were enrolled in this study, of whom 59.3% were IgM positive and 96.4% were IgG positive. The multiple linear regression analysis revealed that the duration of RNA shedding, C-reactive protein level and disease severity were independent predictors of IgG levels. Conclusion: COVID-19 patients retained long-term viral-specific protective immunity. Disease severity, C-reactive protein level and duration of RNA shedding were related to antibody levels 8 months after symptom onset.

Clinical and diagnostic value of the combination of lymphocyte count and creatine kinase in the detection of coronavirus 2019.

The present study aimed to investigate the diagnostic efficiency of the absolute number of lymphocytes (LYM) and creatine kinase (CK) levels in the diagnosis of coronavirus disease 2019 (COVID-19). For this, the clinical data from 84 patients with COVID-19 admitted to Tianjin Haihe Hospital (Tianjin, China) between January and February 2020 were collected. The patients were divided into the following groups: The common COVID-19 group (n=61) and severe COVID-19 group (n=23). In addition, 30 healthy subjects were included as a control group. The results demonstrated that the percentage of neutrophils (NEU%) was significantly increased, while the absolute number of white blood cells, LYM and the percentage of lymphocytes (LYM%) were significantly decreased in patients with COVID-19. Furthermore, in the severe group, the absolute number of red blood cells in female patients, the NEU%, the neutrophil-to-lymphocyte ratio (NLR) and the serum levels of interleukin-6 and C-reactive protein (CRP) were markedly elevated, while those of LYM and LYM% were significantly decreased (all P<0.05). In addition, in the receiver operating characteristics curve analysis for the combination of LYM + CK, the area under the curve values were 0.96 and 1.00, with a sensitivity of 95.08 and 100%, specificity of 86.67 and 100% and cut-off values of 0.42 and 0.50 for the common and severe COVID-19 group, respectively. The results indicated that the diagnostic efficiency of LYM + CK was higher than that of each single factor. Finally, a moderate correlation of lactate dehydrogenase with CRP and NLR (r=0.492 and 0.433, respectively; both P<0.05) was obtained. Overall, the results of the present study indicated that the values of LYM and CK were associated with the progression of COVID-19, suggesting that the combination of both factors may be of clinical diagnostic value for COVID-19.

Preliminary Analysis of B- and T-Cell Responses to SARS-CoV-2.

BACKGROUND AND OBJECTIVE: Without a specific antiviral treatment or vaccine, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has caused a global pandemic, affecting over 200 countries worldwide. A better understanding of B- and T-cell immunity is critical to the diagnosis, treatment and prevention of coronavirus disease 2019 (COVID-19). METHODS: A cohort of 129 patients with COVID-19 and 20 suspected cases were enrolled in this study, and a lateral flow immunochromatographic assay (LFIA) and a magnetic chemiluminescence enzyme immunoassay (MCLIA) were evaluated for SARS-CoV-2 IgM/IgG detection. Additionally, 127 patients with COVID-19 were selected for the detection of IgM and IgG antibodies to SARS-CoV-2 to evaluate B-cell immunity, and peripheral blood lymphocyte subsets were quantified in 95 patients with COVID-19 to evaluate T-cell immunity. RESULTS: The sensitivity and specificity of LFIA-IgM/IgG and MCLIA-IgM/IgG assays for detecting SARS-CoV infection were > 90%, comparable with reverse transcription polymerase chain reaction detection. IgM antibody levels peaked on day 13 and began to fall on day 21, while IgG antibody levels peaked on day 17 and were maintained until tracking ended. Lymphocyte and subset enumeration suggested that lymphocytopenia occurred in patients with COVID-19. CONCLUSIONS: LFIA-IgM/IgG and MCLIA-IgM/IgG assays can indicate SARS-CoV-2 infection, which elicits an antibody response. Lymphocytopenia occurs in patients with COVID-19, which possibly weakens the T-cell response.

Investigation of nicotinamide as more than an anti-phosphorus drug in chronic hemodialysis patients: a single-center, double-blind, randomized, placebo-controlled trial.

BACKGROUND: Hyperphosphatemia is a common complication of late-stage chronic kidney disease (CKD). Nicotinamide (NAM) has been reported as an adjunctive therapy for hyperphosphatasemia, but the effect of NAM on fibroblast growth factor 23 (FGF23) and Klotho has rarely been reported. METHODS: We randomly assigned 98 patients who underwent regular hemodialysis to received NAM (0.5-1.5 g per day, or 1-3 tablets per day) or placebo (1-3 tablets per day) as an add-on therapy of calcium-based phosphorus binders in a 1:1 ratio. All enrollments were followed-up for 52 weeks. We investigated the serum phosphorus as the primary outcome and serum FGF23 and Klotho as the secondary outcomes. Abdominal aortic calcification (AAC), which had a good correlation with coronary calcification was also compared between the two groups. RESULTS: In total, 37 patients in the placebo group and 35 patients in the NAM group completed the 52-week follow-up. Compared with the placebo group, the NAM group showed a significant decrease of serum phosphorus at the 8th, 12th, 20th, 44th, and 52nd week. There was a declining trend of FGF23 and Klotho in both the placebo and NAM groups. Linear mixed models (LMMs) for overall comparisons by repeated measures of analysis of variance (ANOVA) revealed a significant decrease of FGF23 and slower declining rate of Klotho in the NAM group. No significant difference of AAC was detected between the two groups (P=0.805). CONCLUSIONS: NAM can not only further decrease the phosphorus level but also reduce the FGF23 level and slow down the descending rate of Klotho in chronic hemodialysis patients.

Gingival metastasis of a mediastinal pulmonary sarcomatoid carcinoma: a case report.

BACKGROUND: Pulmonary sarcomatoid carcinoma (PSC) is a rare malignancy with both epithelial and sarcoma components, and high tumor metastasis potential. CASE PRESENTATION: A 63-year-old male patient had a tumor in the right posterior mediastinum, and was eventually diagnosed with PSC and gingival metastasis. The patient underwent thoracoscopic right upper pneumonectomy with lymph node dissections, and the subsequent gingival biopsy revealed a metastatic PSC. The immunohistochemistry revealed that both PSC site tissues were positive for vimentin, CKAE1/AE3 and Ki-67. The patient received radiotherapy and chemotherapy after surgery, and deceased two months later due to systemic tumor metastases. CONCLUSION: PSC metastasis is variable, and leads to diagnostic dilemma or erroneous diagnosis. A differential diagnosis can help to distinguish it from gingival cancer.

Fatty Acid Metabolism is Associated With Disease Severity After H7N9 Infection.

BACKGROUND: Human infections with the H7N9 virus could lead to lung damage and even multiple organ failure, which is closely associated with a high mortality rate. However, the metabolic basis of such systemic alterations remains unknown. METHODS: This study included hospitalized patients (n = 4) with laboratory-confirmed H7N9 infection, healthy controls (n = 9), and two disease control groups comprising patients with pneumonia (n = 9) and patients with pneumonia who received steroid treatment (n = 10). One H7N9-infected patient underwent lung biopsy for histopathological analysis and expression analysis of genes associated with lung homeostasis. H7N9-induced systemic alterations were investigated using metabolomic analysis of sera collected from the four patients by using ultra-performance liquid chromatography-mass spectrometry. Chest digital radiography and laboratory tests were also conducted. FINDINGS: Two of the four patients did not survive the clinical treatments with antiviral medication, steroids, and oxygen therapy. Biopsy revealed disrupted expression of genes associated with lung epithelial integrity. Histopathological analysis demonstrated severe lung inflammation after H7N9 infection. Metabolomic analysis indicated that fatty acid metabolism may be inhibited during H7N9 infection. Serum levels of palmitic acid, erucic acid, and phytal may negatively correlate with the extent of lung inflammation after H7N9 infection. The changes in fatty acid levels may not be due to steroid treatment or pneumonia. INTERPRETATION: Altered structural and secretory properties of the lung epithelium may be associated with the severity of H7N9-infection-induced lung disease. Moreover, fatty acid metabolism level may predict a fatal outcome after H7N9 virus infection.

[Effects and mechanism of drug serum of prepared radix polygoni multiflori on rat osteoblast in vitro].

OBJECTIVE: To study the effect of drug serum of prepared Radix Polygoni Multiflori on rat osteoblast (Ob) and its mechanism. METHODS: The animal serum was prepared by serum pharmacology means. The cells were getting by separating and inducing the SD neonatal rat skull bone. The proliferation and differentiation of Ob were detected by CCK-8, alkaline phosphatase (ALP) activity analysis. And RT-PCR method was used to determine the osteogenesis-related genes expression. RESULTS: Compared with control group, the groups with drug serum of prepared Radix Polygoni Mutiflori, 10%, 20% and 30% had an effect on promotion the proliferation significantly on Ob (P < 0.01). There was no concentration-related manner among groups. The 5% and 10% drug serum decreased ALP activity at the post-translation phase compared with control group, but higher doses (20% and 30%) did not have the same effect. However, drug serum of PR/MIN increased significantly osteogenesis-related genes (OC, ALP, Cbfalpha1) mRNA expression (P < 0.05). CONCLUSION: The drug serum of prepared Radix Polygoni Multiflori can stimulate osteoblast proliferation in vitro, and its mechanism may be associated with increasing osteogenesis-related genes expression.

[Study on extraction process for the icariin of hugu capsule with ultrasonic technology].

OBJECTIVE: To study the best ultrasonic technology of the extraction of icariin of Hugu capsule. METHODS: Used the content of icariin as index, orthogonal experiment was carried out to investigate 4 influential factors as follows: the ultrasonic power (A), the ultrasonic frequency (B), the material fluid ratio (C), the time (D). RESULTS: The best extraction conditions were as follows: the ultrasonic power was 120 W, the ultrasonic frequency was 28 KHz, solid-liquid ratio of 1 : 35, the extraction time was 10 min. CONCLUSION: Optimization of extracting process is simple, quick and low energy consumption. Under these conditions, the extraction of icariin is 1.8 times higher than that of the traditional extraction method.

Characterization and expression of an actin-depolymerizing factor from Eimeria tenella.

Actin-depolymerizing factor (ADF) plays an important role in remodeling the actin cytoskeleton which contributes much to the invasion of host cells by the apicomplexan parasite. The gene encoding for Eimeria tenella ADF with one intron was cloned and identified by the E. tenella genome raw sequence data ( http://www.sanger.ac.uk/projects/E.tenella/ ). The deduced polypeptide sequence was only composed of 118 amino acids (13.14 kDa) without signal peptide and nuclear localization sequence. The amino acid sequence was most similar to the ADF of Toxoplasma gondii, 69.1%. Compared the putative three-dimensional structures between E. tenella and yeast, the actin filament binding sites on the segment from the alpha4-helices to the C-terminal were mostly missed in E. tenella. Real-time RT-PCR and dot blot both revealed that ADF expression was relatively stronger in the sporozoites and merozoites than in sporulated and unsporulated oocysts in both mRNA and protein levels. Northern blot analysis suggested that there was only one form of ADF transcripts in all different life stages of E. tenella. Actin-binding experiment showed that the recombinant fusion ADF protein could bind with actin, which indicated that ADF probably plays an important role in the invasion host of E. tenella.