Mutational signature and prognosis in adenocarcinoma of the bladder.

Adenocarcinoma of the bladder is a rare urinary bladder carcinoma with limited therapy options due to lack of molecular characterization. Here, we aimed to reveal the mutational and transcriptomic landscapes of adenocarcinoma of the bladder and assess any relationship with prognosis. Between February 2015 and June 2021, a total of 23 patients with adenocarcinoma of the bladder were enrolled. These included 16 patients with primary bladder adenocarcinomas and seven patients with urachal adenocarcinoma. Whole exome sequencing (16 patients), whole genome sequencing (16 patients), bulk RNA sequencing (RNA-seq) (19 patients), and single-cell RNA-seq (5 patients) were conducted for the specimens. Correlation analysis, survival analysis, and t-tests were also performed. Prevalent T>A substitutions were observed among somatic mutations, and major trinucleotide contexts included 5'-CTC-3' and 5'-CTG-3'. This pattern was mainly contributed by COSMIC signature 22 related to chemical carcinogen exposure (probably aristolochic acid), which has not been reported in bladder adenocarcinoma. Moreover, genes with copy number changes were also enriched in the KEGG term 'chemical carcinogenesis'. Transcriptomic analysis suggested high immune cell infiltration and luminal-like features in the majority of samples. Interestingly, a small fraction of samples with an APOBEC-derived mutational signature exhibited a higher risk of disease progression compared with samples with only a chemical carcinogen-related signature, confirming the molecular and prognostic heterogeneity of bladder adenocarcinoma. This study presents mutational and transcriptomic landscapes of bladder adenocarcinoma, and indicates that a chemical carcinogen-related mutational signature may be related to a better prognosis compared with an APOBEC signature in adenocarcinoma of the bladder. © 2024 The Pathological Society of Great Britain and Ireland.

Clinical characteristics of IgG4-related disease with lung involvement in 14 patients. / 14例累及肺部的IgG4ç›¸å ³æ€§ç–¾ç— çš„ä¸´åºŠç‰¹å¾.

OBJECTIVES: Immunoglobulin G4-related diseases (IgG4-RD) is a rare autoimmune disease, and there is no specific diagnostic test for patients with lung involvement yet. This study aims to summarize the clinical characteristics of IgG4-RD with lung involvement and improve the understanding and diagnosis of this disease. METHODS: All patients diagnosed with IgG4-RD in the Second Xiangya Hospital from December 2014 to February 2022 were re-diagnosed according to the recommendations of Chinese Expert Consensus on the Diagnosis and Treatment of IGG4-Related Diseases in 2021. The clinical data of 14 IgG4-RD patients with pulmonary abnormalities were collected and analyzed. RESULTS: Among the 14 patients, 11 were males and 3 were females, and the median age of diagnosis was 66 (22-82) years old. Six cases had respiratory symptoms such as cough, sputum and short breath. Extrapulmonary involvement was the most common in the glands of head and neck (6/14), followed by pancreas and bile duct (4/14). Elevated serum IgG4 level was found in all patients, and most (11/14) were accompanied by abnormal inflammatory markers. Patients' pulmonary imaging findings were diverse, the most common performances were mediastinal/hilar lymphadenopathy (12/14), followed by multiple pulmonary nodules (9/14), patchy density enhancement (7/14) and the increased broncho vascular bundles (6/14). Lung biopsy was performed in 9 patients, their pathology results showed lymphoplasmic cell infiltration, 5 cases of them had interstitial fibrosis, 2 cases with phlebitis, and extrapulmonary biopsy was performed in 8 patients. Immunohistochemical results of all the patients showed that the number of IgG4+ plasma cells was more than 10 per high magnification, and the ratio of IgG4/IgG was more than 40%. For treatment, 12 patients received hormone therapy, and 5 patients combined immunosuppressive therapy with hormone. 10 patients were in remission after treatment, while 2 patients were progressed. CONCLUSIONS: IgG4-RD with lung involvement is rare and has no specific clinical manifestation. Its pulmonary imaging is diverse. Diagnosis for it should combine with serum IgG4 level and pathological examination. Glucocorticoid is the first line treatment, and combination with immunosuppressant can help prevent disease recurrence.

Clinical characteristics of 32 cases of diffuse panbronchiolitis. / 32例弥漫性泛细支气管炎的临床特征.

OBJECTIVES: Diffuse panbronchiolitis (DPB) is a chronic airway inflammation with low specificity and its diagnosis is often missed or delayed. This study aims to summarize the clinical characteristics and treatment of DPB in order to improve the understanding and diagnosis of the disease. METHODS: The clinical data of 32 DPB patients were collected, analyzed and summarized from March 1, 2013 to March 1, 2022 in the Second Xiangya Hospital of Central South University. The basic information, clinical manifestations, laboratory tests, pulmonary function, imaging tests, treatment, and regression of patients were analyzed. RESULTS: A total of 32 patients were enrolled in the final analysis, with a male-to-female ratio at 1.67. The median age at symptom onset was 26.5 (11.0-69.0) years, and the median age of diagnosis was 47.5 (16.0-77.0) years. All patients presented with chronic cough and copious sputum production. A total of 26 patients had post activity shortness of breath and 14 patients had a positive result (blood cold agglutination test titer≥1꞉64). Pulmonary function examination was performed in 31 patients, 18 patients showed mixed pulmonary ventilation dysfunction, 12 patients showed obstructive pulmonary ventilation, and 1 patient had normal pulmonary ventilation function. A total of 31 patients had a bilateral, diffuse, small nodule pattern on chest CT. All patients were treated with macrolides. A total of 31 patients showed improvement, and 20 patients showed improvement in partial pressure of oxygen and blood oxygen saturation compared with before at discharge. A total of 12 patients were re-examined by chest CT after completing macrolides treatment, 6 cases showed less diffuse nodules, 5 cases showed no significant changes, and 1 case showed more diffuse nodules, which indicated the disease progression. Seven patients received pulmonary function tests after completing macrolides treatment, forced expiratory volume in one second (FEV1) and FEV1/forced vital capacitywere improved, but forced expiratory flow at 25% of vital capacity did not change significantly. CONCLUSIONS: The clinical manifestations of DPB are nonspecific. Early diagnosis and treatment are very important for the prognosis of patients.

Drug Resistance (Dapsone, Rifampicin, Ofloxacin) and Resistance-Related Gene Mutation Features in Leprosy Patients: A Systematic Review and Meta-Analysis.

Dapsone (DDS), Rifampicin (RIF) and Ofloxacin (OFL) are drugs recommended by the World Health Organization (WHO) for the treatment of leprosy. In the context of leprosy, resistance to these drugs occurs mainly due to mutations in the target genes (Folp1, RpoB and GyrA). It is important to monitor antimicrobial resistance in patients with leprosy. Therefore, we performed a meta-analysis of drug resistance in Mycobacterium leprae and the mutational profile of the target genes. In this paper, we limited the study period to May 2022 and searched PubMed, Web of Science (WOS), Scopus, and Embase databases for identified studies. Two independent reviewers extracted the study data. Mutation and drug-resistance rates were estimated in Stata 16.0. The results demonstrated that the drug-resistance rate was 10.18% (95% CI: 7.85-12.51). Subgroup analysis showed the highest resistance rate was in the Western Pacific region (17.05%, 95% CI:1.80 to 13.78), and it was higher after 2009 than before [(11.39%, 7.46-15.33) vs. 6.59% (3.66-9.53)]. We can conclude that the rate among new cases (7.25%, 95% CI: 4.65-9.84) was lower than the relapsed (14.26%, 95 CI%: 9.82-18.71). Mutation rates of Folp1, RpoB and GyrA were 4.40% (95% CI: 3.02-5.77), 3.66% (95% CI: 2.41-4.90) and 1.28% (95% CI: 0.87-1.71) respectively, while the rate for polygenes mutation was 1.73% (0.83-2.63). For further analysis, we used 368 drug-resistant strains as research subjects and found that codons (Ser, Pro, Ala) on RpoB, Folp1 and GyrA are the most common mutation sites in the determining region (DRDR). In addition, the most common substitution patterns of Folp1, RpoB, and GyrA are Pro→Leu, Ser→Leu, and Ala→Val. This study found that a higher proportion of patients has developed resistance to these drugs, and the rate has increased since 2009, which continue to pose a challenge to clinicians. In addition, the amino acid alterations in the sequence of the DRDR regions and the substitution patterns mentioned in the study also provide new ideas for clinical treatment options.

Effects of 25-hydroxyvitamin D3 on growth performance, serum parameters, fecal microbiota, and metabolites in weaned piglets fed diets with low calcium and phosphorus.

BACKGROUND: With the development of intensive farming, long-term exposure of pigs to poor light conditions is not conducive to the production of vitamin D3 , and vitamin D3 deficiency could affect absorption and metabolism of calcium (Ca) and phosphorus (P). 25-Hydroxyvitamin D3 (25OHD3 ) has higher bioactivity than regular vitamin D3 . This study investigated the effects of 25OHD3 on performance, serum parameters, fecal microbiota, and metabolites in weaned piglets fed with low Ca-P diet. RESULTS: It was found that a low Ca-P diet supplemented with 50 µg/kg 25OHD3 (NC + 25-D) improved (P < 0.05) average daily gain (ADG) in phase 2 and in the overall period of the experiment, and increased (P < 0.05) the immunoglobulin G (IgG), immunoglobulin A (IgA), catalase (CAT), bone-specific alkaline phosphatase (BALP), and osteocalcin (OC) serum content on day 28 compared with a low Ca-P diet (NC), but no differences were observed between a normal Ca-P diet (PC) and the NC + 25-D diet. Compared with NC, the abundance of Firmicutes was higher (P < 0.05) in PC and NC + 25-D. NC + 25-D decreased (P < 0.05) the abundance of Streptococcaceae compared with PC and NC, and increased (P < 0.05) the abundance of Lachnospiraceae compared with NC. Serum 25OHD3 was negatively correlated with the abundance of fecal Streptococcaceae (P < 0.05), and positively correlated with the abundance of fecal Lachnospiraceae (P < 0.05). CONCLUSION: Supplementation of 25OHD3 in a low Ca-P diet improved serum immunity, bone biochemical parameters, and fecal microbiota such as decreased Streptococcaceae abundance and increased Lachnospiraceae abundance, which could subsequently promote growth of piglets. The effects were similar to that of a normal Ca-P diet. © 2021 Society of Chemical Industry.

Clinical analysis for 15 patients with pulmonary Langerhans cell histiocytosis and literature review. / 15ä¾‹æˆäººè‚ºæœ—æ ¼æ±‰æ–¯ç»†èƒžç»„ç»‡ç»†èƒžå¢žç”Ÿç—‡ä¸´åºŠåˆ†æžåŠæ–‡çŒ®å›žé¡¾.

OBJECTIVES: Pulmonary Langerhans cell histiocytosis (PLCH) is a clonal disease, characterized by proliferation of Langerhans cells that derived from bone marrow infiltrating the lungs and other organs. Due to the rarity of the disease, the current understanding of the disease is insufficient, often misdiagnosed or missed diagnosis. This study aims to raise clinicians' awareness for this disease via summarizing the clinical characteristics, imaging features, and treatment of PLCH. METHODS: We retrospectively analyzed clinical and follow-up data of 15 hospitalized cases of PLCH from September 2012 to June 2021 in the Second Xiangya Hospital of Central South University. RESULTS: The age of 15 patients (9 men and 6 women, with a sex ratio of 3 to 2) was 21-52 (median 33) years. Among them, 8 had a history of smoking and 5 suffered spontaneous pneumothorax during disease course. There were 3 patients with single system PLCH and 12 patients with multi-system PLCH, including 7 patients with pituitary involvement, 7 patients with lymph node involvement, 6 patients with bone involvement, 5 patients with liver involvement, 2 patients with skin involvement, 2 patients with thyroid involvement, and 1 patients with thymus involvement. The clinical manifestations were varied but non-specific. Respiratory symptoms mainly included dry cough, sputum expectoration, chest pain, etc. Constitutional symptoms included fever and weight loss. Patients with multi-system involvement experienced symptoms such as polyuria-polydipsia, bone pain, and skin rash. All patients were confirmed by pathology, including 6 by lung biopsy, 3 by bone biopsy, 2 by lymph node biopsy, and 4 by liver, skin, suprasternal fossa tumor, or pituitary stalk biopsy. The most common CT findings from this cohort of patients were nodules and/or cysts and nodular and cystic shadows were found in 7 patients. Three patients presented simple multiple cystic shadows, 3 patients presented multiple nodules, and 2 patients presented with single nodules and mass shadows. Pulmonary function tests were performed in 4 patients, ventilation dysfunction was showed in 2 patients at the first visit. Pulmonary diffusion function tests were performed in 4 patients and showed a decrease in 3 patients. Smoking cessation was recommended to PLCH patients with smoking history. Ten patients received chemotherapy while 2 patients received oral glucocorticoid therapy. Among the 11 patients with the long-term follow-up, 9 were in stable condition. CONCLUSIONS: PLCH is a neoplastic disease closely related to smoking. The clinical manifestations and laboratory examination are not specific. Pneumothorax could be the first symptom which is very suggestive of the disease. Definitive diagnosis relies on histology. There is no unified treatment plan for PLCH, and individualized treatment should be carried out according to organ involvement. Early smoking cessation is essential. Chemotherapy is the main treatment for rapidly progressing PLCH involved multiple organs. All diagnosed patients can be considered for the detection of BRAFV600E gene and relevant targeted therapies have been implemented recently.

Clinical characteristics of eosinophilic granulomatosis with polyangiitis involving the lung in 13 patients. / 13ä¾‹ç´¯åŠè‚ºéƒ¨çš„å—œé ¸æ€§è‚‰èŠ½è‚¿æ€§å¤šè¡€ç®¡ç‚Žçš„ä¸´åºŠç‰¹å¾.

OBJECTIVES: Eosinophilic granulomatosis with polyangiitis (EGPA) is a rare systemic vasculitis, which often starts with respiratory symptoms such as asthma, and it is difficult to make early clinical diagnosis.This study aims to improve the therapeutic level of EGPA with lung involvement via analyzing the clinical characteristics, diagnosis, and treatment . METHODS: We retrospectively analyzed the clinical data of 13 EGPA patients with lung involvement who were diagnosed from February 1, 2014 to July 31, 2021 in the Second Xiangya Hospital, Central South University. RESULTS: The ratio of male to female in 13 patients was 7꞉6. The patients were diagnosed at median age 52 (46-68) years old and 6 had been diagnosed as "bronchial asthma". Pulmonary clinical manifestations mainly included cough, expectoration, wheezing, and shortness of breath; while extra-pulmonary manifestations mainly included rash and subcutaneous mass, fever, limb numbness, muscle and joint pain, abdominal pain, etc. Peripheral blood tests of all patients showed that 11 patients had eosinophils ≥10%, 10 had elevated inflammatory indicators, and 3 were anti-neutrophil cytoplasmic antibody (ANCA) positive. The major lung imaging features were patches or strips of increased density, multiple nodules, bronchiectasis, bronchial wall thickening, exudation, mediastinal lymph nodes, and so on. Eight patients had sinusitis and 9 with abnormal electromyography. Extravascular eosinophil infiltration was found in 9 patients. Six patients with lung biopsy showed eosinophil, lymphocyte, and plasma cell infiltration, 3 patients were involved in small blood vessels, and 1 had granuloma. Pulmonary function tests were performed in 7 patients, 5 of them showed different degrees of pulmonary ventilation dysfunction, and 4 of them had diffusion dysfunction. Almost all patients respond well to glucocorticoid and immunosuppressant. CONCLUSIONS: EGPA is rare in clinical, often involving multiple systems with great harm and may combine with asthmatic manifestations. Pulmonary involvement is relatively common. However, due to insufficient recognition of this disease and huge heterogeneity of pulmonary imaging manifestations, misdiagnosis and missed diagnosis are easy to occur. Relevant laboratory, imaging, and biopsy examination should be performed as early as possible with comprehensive consideration of extrapulmonary involvement. Early identification has great significance to improve the diagnosis rate and prognosis of diseases.

Single-cell RNA sequencing reveals the epithelial cell heterogeneity and invasive subpopulation in human bladder cancer.

Bladder cancer represents a highly heterogeneous disease characterized by distinct histological, molecular and clinical phenotypes, and a detailed analysis of tumor cell invasion and crosstalks within bladder tumor cells has not been determined. Here, we applied droplet-based single-cell RNA sequencing (scRNA-seq) to acquire transcriptional profiles of 36 619 single cells isolated from seven patients. Single cell transcriptional profiles matched well with the pathological basal/luminal subtypes. Notably, in T1 tumors diagnosed as luminal subtype, basal cells displayed characteristics of epithelial-mesenchymal transition (EMT) and mainly located at the tumor-stromal interface as well as micrometastases in the lamina propria. In one T3 tumor, muscle-invasive tumor showed significantly higher expression of cancer stem cell markers SOX9 and SOX2 than the primary tumor. We additionally analyzed communications between tumor cells and demonstrated its relevance to basal/luminal phenotypes. Overall, our single-cell study provides a deeper insight into the tumor cell heterogeneity associated with bladder cancer progression.

Dietary 25-hydroxycholecalciferol supplementation improves performance, immunity, antioxidant status, intestinal morphology, and bone quality in weaned piglets.

BACKGROUND: 25-Hydroxycholecalciferol (25OHD3 ) is a new feed additive, which is a potential alternative to vitamin D3 in swine nutrition. The objective of this study was to determine the effects of different doses of 25OHD3 supplementation on performance, immunity, antioxidant capacity, intestinal morphology and bone quality in piglets. RESULTS: As dietary 25OHD3 supplementation increased, the average daily gain (ADG) improved (P < 0.05) quadratically during days 1-14, and tended to increase (P = 0.06) quadratically during the overall period of the experiment. Increasing 25OHD3 supplementation increased (linear effect, P < 0.05) the serum 25OHD3 level and serum glutathione peroxidase (GSH-Px) activity. On day 14, serum immunoglobulin A (IgA) was increased (linear and quadratic effects, P < 0.05) as dietary 25OHD3 supplementation increased. On day 28, serum IgA level was higher (P < 0.05) linearly and the complement 3 (C3) level was reduced (P < 0.05) linearly as dietary supplementation of 25OHD3 increased. The mucosal GSH-Px activity of the small intestine was higher (quadratic effect, P < 0.05) with increasing 25OHD3 supplementation. Jejunal villus height (P = 0.06) and villus height to crypt depth ratio (P = 0.07) tended to increase quadratically, and the villus height to crypt-depth ratio of the ileum increased (P < 0.05) linearly and quadratically with increasing 25OHD3 supplementation. Dietary supplementation with an increasing level of 25OHD3 increased breaking strength of tibias and femurs (quadratic effect, P < 0.05). CONCLUSION: Increasing dietary 25OHD3 supplementation partly improved performance, immunity, antioxidant status, intestinal morphology, and bone properties of weaned piglets. © 2020 Society of Chemical Industry.

Effects of maternal 25-hydroxycholecalciferol during the last week of gestation and lactation on serum parameters, intestinal morphology and microbiota in suckling piglets.

This study was conducted to test the effects of maternal 25-hydroxycholecalciferol (25OHD3) supplementation on serum parameters, intestinal morphology and microbiota in suckling piglets. The experiment started on day 107 of gestation and lasted until piglets were weaned on day 21 of lactation. Thirty-two sows were allocated randomly to two treatments (ND diet, basal diet with 2000 IU/kg of vitamin D3; 25-D diet, basal diet with 50 µg/kg 25OHD3). Results showed that maternal 25-D treatment increased (p < 0.05) serum 25OHD3 concentration in the umbilical cords, which led to higher (p < 0.05) serum 25OHD3 concentration of suckling piglets from 25-D sows. The GSH-Px activity in colostrum was higher (p < 0.05), as well as SOD and GSH-Px activities in milk, were higher (p < 0.05) in 25-D sows than ND sows. Compared with piglets suckling ND sows, piglets suckling 25-D sows had higher (p < 0.05) serum SOD activity on day 7, 14 and 21 of lactation. On day 21 of lactation, piglets form 25-D sows had greater (p < 0.05) serum levels of GH and IGF-I and lower (p < 0.05) serum DAO activity than those from ND sows. Piglets from 25-D sows had higher (p < 0.05) jejunal villus height than those from ND sows. Feeding 25OHD3 to sows tended to increase (p < 0.10) the species richness in the colonic digesta of suckling piglets, as reflected by the α-diversity index of Chao-1. In the caecal digesta, the α-diversity for bacterial community analysis of Simpson and Shannon was lower (p < 0.05) in 25-D piglets than ND piglets. The relative abundances of colonic Alloprevotella and caecal Lactobacillus were significantly higher, while the population of caecal [Eubacterium]_coprostanoligenes_group was lower (p < 0.05) in 25-D piglets than ND piglets. In conclusion, maternal 25OHD3 supplementation partly improved antioxidant status in sows and suckling piglets and altered gut microbiota in the hindgut of piglets.

Stable and Photothermally Efficient Antibody-Covered Cu3(PO4)2@Polydopamine Nanocomposites for Sensitive and Cost-Effective Immunoassays.

Polydopamine (PDA)-coated or encapsulating Cu3(PO4)2 (Cu3(PO4)2@PDA) nanosheets were synthesized, allowing the C-reaction protein (CRP) antibody to be attached electrostatically for immunosensing of CRP with simple photothermal detection. The antibody-covered Cu3(PO4)2@PDA nanosheets replace the antibody-conjugated enzyme in the enzyme-linked immunosorbant assays. Owing to the high surface area of the 2-D-structured Cu3(PO4)2@PDA nanosheets and the coabsorption of light in the near-IR spectrum by Cu3(PO4)2 and PDA, a small amount of Cu3(PO4)2@PDA confined in the wells of a titer plate generates an easily detectable temperature change after irradiation at 808 nm. The temperature changes, measured by an inexpensive pen-type thermometer, increased linearly with the analyte concentration from 0.42 to 16 pM. We found that the linear relationship can be fitted by the isotherm derived from responses collected from heterogeneous sensors covered with different ligand or antibody densities. The low detection limit (0.11 pM) is largely due to the attachment of a great number of antibodies onto the flat nanosheets. The antibody-covered Cu3(PO4)2@PDA nanosheets are stable and can be used under conditions that are generally unfavorable to enzymatic activities. The excellent agreement between our results and immunoturbidimetric assays of CRP in serum samples from patients and healthy donors demonstrates its utility for disease diagnosis in clinical settings. This cost-effective, biocompatible, and convenient photothermal immunosensor affords a range of possibilities for detecting diverse protein biomarkers.

CD163+ macrophages predict a poor prognosis in patients with primary T1 high-grade urothelial carcinoma of the bladder.

PURPOSE: Macrophages are a major cell type that can infiltrate solid tumors and exhibit distinct phenotypes in different tumor microenvironments. This study investigates the prognostic value of tumor-infiltrated CD163+ macrophages in patients with T1 high-grade (T1HG) bladder cancer. METHODS: CD163+ macrophages were assessed by immunohistochemistry in 94 T1HG bladder cancer samples. Kaplan-Meier analyses and Cox proportional hazards' regression models were applied to evaluate recurrence-free survival, progression-free survival and disease-specific survival. RESULTS: With a median follow-up of 60 months, 37 (39.4%) patients experienced disease recurrence, 14 (14.9%) progression, 11 (11.7%) disease-specific mortality. High CD163+ macrophages were associated with higher risk of disease recurrence and progression (P < 0.05, for both). In multivariate Cox proportional hazards regression analysis, high CD163+ macrophages were a significant negative predictor of recurrence-free survival, progression-free survival and disease-specific survival (P < 0.05 for all). CONCLUSION: CD163+ macrophages are a poor prognostic factor in T1HG bladder cancer. This finding provide the ground for further testing it in predicting the outcome of this challenging disease.

Maternal 25-hydroxycholecalciferol during lactation improves intestinal calcium absorption and bone properties in sow-suckling piglet pairs.

Lower maternal vitamin D status during lactation is a common health problem. The objectives of this study were to investigate the effects of maternal 25-hydroxycholecalciferol (25-OH-D3) supplementation during lactation on maternal and neonatal bone health in a sow model. 32 Large White × Landrace sows were assigned randomly to one of two diets supplemented with 2000 IU/kg vitamin D3 (ND) or 50 µg/kg 25-OH-D3 (25-D). The experiment began on day 107 of gestation and continued until weaning on day 21 of lactation. Maternal 25-OH-D3 supplementation significantly decreased milk n-6:n-3 PUFA ratio, which supported bone formation of piglets. Supplementation with 25-OH-D3 altered bone turnover rate of sows and piglets, as evidenced by higher bone-specific alkaline phosphatase (BALP) concentration in serum. 25-D sows had significantly higher bone density and mechanical properties of tibias and femurs than ND sows. Calcium (Ca) absorption rate was higher in 25-D sows than ND sows, which was caused partially by the increased mRNA expressions of renal 1α-hydroxylase (CYP27B1) and duodenal vitamin D receptor (VDR), transient receptor potential vanilloid 6 (TRPV6), and calcium-binding protein D9k (CaBP-D9k). Maternal 25-OH-D3 supplementation increased tibial and femoral Ca content by up-regulating Ca-related gene expression in kidney (CYP27B1), ileum (VDR and claudin-2), and colon (VDR and CaBP-D9k), thus, activating 1,25-dihydroxyvitamin D3 [1,25-(OH)2-D3]-dependent Ca transport in piglets. In conclusion, improved milk fatty acids and higher mRNA expressions of calcitropic genes triggered by maternal 25-OH-D3 supplementation would be the potential mechanism underlying the positive effects of 25-OH-D3 on maternal and neonatal bone health.

Ratiometric fluorescent immunoassay for the cardiac troponin-I using carbon dots and palladium-iridium nanocubes with peroxidase-mimicking activity.

A nanozyme-linked immunosorbent assay is described for cardiac troponin I which is a biomarker for myocardial infarction. The method is based on the use of Pd-Ir nanocubes with excellent peroxidase-like activity. The nanocubes catalyze the oxidization of nonfluorescent o-phenylenediamine (OPD) by H2O2 to form a yellow fluorescent product (oxOPD) with excitation/emission maxima at 400/570 nm. Carbon dots are added as a reference fluorophore. Under the same excitation wavelength, they display blue fluorescence (450 nm). The ELISA uses the Pd-Ir nanocubes as a label for the secondary antibody and OPD as substrate. The ratio of fluorescence intensities at 570 and 450 nm increases in the 1 pg·mL-1 to 1 ng·mL-1 cardiac troponin I concentration range, and the detection limit is 0.31 pg·mL-1. The method was applied to analyze spiked serum samples, and the results compared well with those obtained by a commercial chemiluminescence assay. Graphical abstract Schematic presentation of the ratiometric fluorescence immunoassay for cardiac troponin-I. Pd-Ir nanocubes were employed to fabricate nanozyme-based signal labels for its excellent peroxidase-mimicking activity.

Effects of dietary calcium and phosphorus levels and supplementation of 25-hydroxycholecalciferol on performance and bone properties of broiler starters.

To investigate effects of dietary calcium (Ca) and phosphorus (P) levels and 25-hydroxycholecalciferol (25OHD3) supplementation on performance and bone properties of broiler starters, 224 male Arbor Acre broilers were used in a 21-d trial. Broilers were allotted to one of four treatments in a 2 × 2 factorial arrangement including diets either normal or low in Ca and P, which were further supplemented or not with 69 µg 25OHD3/kg feed. Feeding low Ca and P diets significantly reduced performance of boilers and decreased ash, Ca, P and hydroxyproline contents in tibias and femurs (p < 0.05). Ultimate load, bending moment, stiffness and energy to fail were decreased (p < 0.05) in broilers fed diets deficient in Ca and P. Addition of 25OHD3 did not influence performance but significantly increased serum 25OHD3 levels. Furthermore, the addition of 25OHD3 caused an increased tibial and femoral bone density and femoral hydroxyproline content (p < 0.05), increased bending moment in tibias (p < 0.05), and enhanced ultimate load and bending moment in femurs (p < 0.05). No significant interactions were observed for bone properties. Overall, feeding 25OHD3 at 69 µg/kg feed to broilers had no effect on growth performance but partly improved bone biochemical and biomechanical properties of broiler starters.

Effects of maternal 25-hydroxycholecalciferol on nutrient digestibility, milk composition and fatty-acid profile of lactating sows and gut bacterial metabolites in the hindgut of suckling piglets.

To investigate the effects of maternal 25-hydroxycholecalciferol (25OHD3) supplementation during lactation on nutrient digestibility and milk composition of sows and gut bacterial metabolites and their metabolites in the hindgut of suckling piglets, 24 Large White × Landrace sows were assigned randomly to one of two dietary treatments (Diet ND: 2000 IU vitamin D3/kg feed; Diet 25-D: 50 µg 25OHD3/kg feed). The experiment began on d 107 of gestation and continued until weaning on d 21 of lactation. Maternal 25OHD3 supplementation increased (p < 0.05) total litter weight gain during lactation. Milk fat content, immunoglobulin G level on d 21 of lactation and 25OHD3 concentration on d 7, 14, and 21 of lactation were higher (p < 0.05) in sows fed with 25OHD3. Apparent total tract digestibility of dietary calcium was higher (p < 0.05) in 25-D sows than ND sows. With respect to fatty-acid profile, C16:0 and saturated fatty acids in milk were higher (p < 0.05), but C20:4n-6, the ratios of monounsaturated fatty acids to saturated fatty acids and polyunsaturated fatty acids to saturated fatty acids were lower (p < 0.05) in 25-D sows than ND sows. 25OHD3 supplementation increased the mRNA expressions of acetyl-CoA carboxylase α and fatty-acid synthase in the mammary gland of lactating sows. For gut bacterial metabolites, concentration of butyrate in the caecal digesta was higher (p < 0.05) in piglets suckling 25-D sows than piglets suckling ND sows. In conclusion, 25OHD3 supplementation in maternal diets changed dietary calcium digestibility, milk composition and milk fatty-acid profile of lactating sows and altered gut bacterial metabolites in the hindgut of suckling piglets.

Mixed organic acids improve nutrients digestibility, volatile fatty acids composition and intestinal microbiota in growing-finishing pigs fed high-fiber diet.

OBJECTIVE: The objective of this study was to investigate effects of mixed organic acids (MOA) on nutrient digestibility, volatile fatty acids composition and intestinal microbiota in growing-finishing pigs fed high wheat bran diet. METHODS: Six crossbred barrows (Duroc×Landrace×Yorkshire), with an average body weight 78.8±4.21 kg, fitted with T-cannulas at the distal ileum, were allotted to a double 3×3 Latin square design with 3 periods and 3 diets. Each period consisted of a 5-d adjustment period followed by a 2-d total collection of feces and then a 2-d collection of ileal digesta. The dietary treatments included a corn-soybean-wheat bran basal diet (CTR), mixed organic acid 1 diet (MOA1; CTR+3,000 mg/kg OA1), mixed organic acid 2 diet (MOA2; CTR+2,000 mg/kg OA2). RESULTS: Pigs fed MOA (MOA1 or MOA2) showed improved (p<0.05) apparent total tract digestibility (ATTD) of gross energy, dry matter and organic matter, and pigs fed MOA2 had increased (p<0.05) ATTD of neutral detergent fiber compared to CTR. Dietary MOA supplementation decreased (p<0.05) pH value, and improved (p<0.01) concentrations of lactic acid and total volatile fatty acids (TVFA) in ileum compared to CTR. Pigs fed MOA showed higher (p<0.05) concentration of acetic acid, and lower (p<0.05) content of formic acid in feces compared to CTR. Pigs fed MOA1 had increased (p<0.05) concentration of TVFA and butyric acid in feces. Pigs fed MOA1 showed higher concentration of Lactobacillus and lower concentration of Escherichia in feces compared to CTR. CONCLUSION: Dietary supplementation of MOA 1 or 2 could improve nutrients digestibility, TVFA concentration and intestinal flora in growing-finishing pigs fed high fiber diet.

Enzyme-Free Immunosorbent Assay of Prostate Specific Antigen Amplified by Releasing pH Indicator Molecules Entrapped in Mesoporous Silica Nanoparticles.

An enzyme-free titer plate-based colorimetric assay utilizing functionalized mesoporous silica nanoparticles (MSNs) entrapping pH-indicator molecules has been developed. Pores in the silica nanoparticles were functionalized with phenyltrimethyloxysilane so that pH indicator molecules (thymolphthalein or TP in the present case) can be tightly entrapped through π-π conjugation. To detect prostate specific antigen (PSA), the TP-containing MSNs were coated with polyethylenimine (PEI), which favors the attachment of the negatively charged secondary anti-PSA antibody. The entrapped thymolphthalein molecules can be readily released from the pores with a simple addition of alkaline solution. The resultant bifunctional MSNs were used for signal-amplified detection of PSA captured by the primary antibody preimmobilized in the wells of a plate. Our method possesses a wide dynamic range (0.5 to 8000 pg/mL) wherein the adsorption of the bifunctional MSNs obeys a modified Langmuir isotherm. A detection limit (LOD) down to as low as 0.36 pg/mL can be attained. Owing to the size uniformity of the MSNs and the obviation of enzyme molecules employed in the enzyme-linked immunosorbent assay (ELISA), excellent reproducibility (RSD = 1.12%) was achieved. The selective detection of PSA in human serum samples demonstrates the amenability of our method to detect important biomarkers in complex biological samples, whereas the performance of the assay in a titer plate ensures high throughput and obviates the use of expensive instruments. Both of these features are prerequisites for clinical settings wherein a great number of samples need to be analyzed in a timely fashion.

Dental health and treatment needs in people with leprosy in China.

OBJECTIVES: To evaluate the dental health status and treatment needs of people affected by leprosy in China, and provide a basis for the development of national or regional dental health programmes to cover the treatment needs of this population. DESIGN: A cross-sectional study with 613 former leprosy patients was carried out in six leprosy villages, in three provinces in China (Nanjing, Taixing and Jiangyan in Jiangsu Province, Hanzhong and Shangluo in Shanxi Province and Yongzhou in Hunan Province). A questionnaire about demographic and clinical data was used. The World Health Organization's (WHO) basic methods were used to determine the tooth-based treatment needs. Periodontal status was determined by using the Community Periodontal Index of Treatment Needs (CPITN). In addition, prosthetic normative needs were assessed. RESULTS: Among the 613 people affected by leprosy, there were 472 people (77%) who had never visited a dentist and 172 people (28.1%) had never brushed their teeth; 302 (49.3%) brushed their teeth once a day. However, there were 267 people (43.6%) who thought their dental health was at an average level and 108 (17.6%) thought they had good dental health. 55.6% of the subjects required dental fillings, 32.7% required pulp care and restoration, and 71.1% required extraction. On CPITN, 23.2% of the subjects scored 2, 28.6% scored three and 48.0% scored four, showing that these people required systematic periodontal treatment. In addition, 84.5% of the subjects needed normative prosthetic treatment. CONCLUSIONS: Most of the subjects with leprosy in this study lacked self-care knowledge on dental health, and especially self-awareness of dental conditions. Normative treatment needs of people affected by leprosy were very high. This result calls for improved oral health education and oral health care in people with leprosy. Oral health education might preferably be integrated into already existing leprosy rehabilitation programs.

Promotion of the Clostridium acetobutylicum ATCC 824 growth and acetone-butanol-ethanol fermentation by flavonoids.

An unexpected promotion effect of Ginkgo leaf on the growth of Clostridium acetobutylicum ATCC 824 and acetone-butanol-ethanol (ABE) fermentation was investigated. Component analysis of Ginkgo leaf was carried out and flavonoids were determined as the potential key metabolites. Then the flavonoids feeding experiments were carried out. Results showed that addition of only 10 mg/L flavonoids to the fermentation broth can promote butanol and ABE titre up to 14.5 and 17.8 g/L after 5 days of fermentation, that is, 74 and 68% higher than the control. A 2.2-fold biomass also has been achieved. Furthermore, by employing such novel founding, we easily exploited flavonoids from soybean and some agriculture wastes as the wide-distributed and economic feasible ABE fermentation promoter. The mechanism of the above effects was investigated from the perspective of oxidation-reduction potential. This work opens a new way in the efforts to increase the titer of butanol.