Prediction and Identification of GPCRs Targeting for Drug Repurposing in Osteosarcoma.

Background: Osteosarcoma (OS) is a malignant bone tumor common in children and adolescents. The 5-year survival rate is only 67-69% and there is an urgent need to explore novel drugs effective for the OS. G protein-coupled receptors (GPCRs) are the common drug targets and have been found to be associated with the OS, but have been seldom used in OS. Methods: The GPCRs were obtained from GPCRdb, and the GPCRs expression profile of the OS was downloaded from the UCSC Xena platform including clinical data. 10-GPCRs model signatures related to OS risk were identified by risk model analysis with R software. The predictive ability and pathological association of the signatures in OS were explored by bio-informatics analysis. The therapeutic effect of the target was investigated, followed by the investigation of the targeting drug by the colony formation experiment were. Results: We screened out 10 representative GPCRs from 50 GPCRs related to OS risk and established a 10-GPCRs prognostic model (with CCR4, HCRTR2, DRD2, HTR1A, GPR158, and GPR3 as protective factors, and HTR1E, OPN3, GRM4, and GPR144 as risk factors). We found that the low-risk group of the model was significantly associated with the higher survival probability, with the area under the curve (AUC) of the ROC greater than 0.9, conforming with the model. Moreover, both risk-score and metastasis were the independent risk factor of the OS, and the risk score was positively associated with the metastatic. Importantly, the CD8 T-cells were more aggregated in the low-risk group, in line with the predict survival rate of the model. Finally, we found that DRD2 was a novel target with approved drugs (cabergoline and bromocriptine), and preliminarily proved the therapeutic effects of the drugs on OS. These novel findings might facilitate the development of OS drugs. Conclusion: This study offers a satisfactory 10-GPCRs model signature to predict the OS prognostic, and based on the model signature, candidate targets with approved drugs were provided.

Resection and total femoral prosthesis reconstruction treatment of massive femoral hemophilic pseudotumor: A case report.

INTRODUCTION AND IMPORTANCE: Hemophilic pseudotumor is a rare and serious complication of hemophilia, often occurring in the long bones and muscle tissue of the extremities, with an incidence of about 1-2%. However, there is no effective surgical treatment for massive hemophilia pseudotumors of the extremities. Therefore, we would like to report this case to provide new ideas and methods for the treatment of massive hemophilic pseudotumors of the extremities through resection and total femoral prosthesis reconstruction. CASE PRESENTATION: After admission, the patient first underwent a month-long coagulation factor replacement therapy to maintain the patient's factor IX level at approximately 100%. Then, extensive resection of hemophilic pseudotumor and total femoral prosthesis reconstruction was performed in collaboration with several departments, and we collected the patient's preoperative, intraoperative and postoperative data for analysis. CLINICAL DISCUSSION: After coagulation factor replacement therapy the patient's factor IX level was significantly increased and approached 100%. The surgical procedure was uneventful, and postoperative X-ray suggested a well-positioned prosthesis with postoperative pathological features consistent with a hemophilic pseudotumor. The patient was able to move around with the assistance of a walker 14 days after surgery, and there was no recurrence after one year of follow-up. CONCLUSION: For treat the massive hemophilic pseudotumors of the extremities, extensive resection and prosthetic replacement with coagulation factor replacement therapy is a safe and effective treatment that can significantly improve the prognosis and quality of life of patients.

The Association Between Inflammatory and Oxidative Stress Biomarkers and Plasma Metabolites in a Longitudinal Study of Healthy Male Welders.

INTRODUCTION: Human metabolism and inflammation are closely related modulators of homeostasis and immunity. Metabolic profiling is a useful tool to understand the association between metabolism and inflammation at a systemic level. OBJECTIVE: To investigate the longitudinal associations between the concentration of plasma metabolites and biomarkers related to inflammation and oxidative stress. METHODS: We conducted a repeated cross-sectional analysis consisting of 8 short-term panels that included 88 healthy adult male welders in Massachusetts, USA. In each panel, we collected 1-6 repeated measurements of blood and urine. We used a human vascular injury panel assay and custom cytokine/chemokine assay to quantify inflammatory biomarker plasma levels, liquid chromatography-mass spectrometry to quantify the concentrations of 665 plasma metabolites, and a competitive enzyme-linked immunoassay to quantify urinary 8-OHdG and 8-isoprostane levels. We used linear mixed effects models to estimate the longitudinal association between each inflammatory and oxidative stress biomarker and each metabolite. RESULTS: At a 5% FDR threshold, we detected ≥1metabolite association for 8 unique inflammatory and oxidative stress biomarkers: urinary 8-isoprostane, plasma C-reactive protein (CRP), serum amyloid A (SAA), intercellular adhesion molecule 1, circulating vascular cell adhesion molecule-1, interleukin 8 (IL-8), interleukin 10 (IL-10) and vascular endothelial growth factor. Specifically, 3 metabolites in the androgenic steroids pathway were negatively associated with SAA; 3 dihydrosphingomyelins metabolites were positively associated with 1 or more of CRP, SAA, IL-8 and IL-10; 4 metabolites in acyl choline metabolism pathways were negatively associated with IL-8; 7 lysophospholipid metabolites were negatively associated with 1 or more of CRP, SAA and IL-8; 4 sphingomyelins were positively associated with CRP and/or SAA; and 10 metabolites in the xanthine pathway were positively associated with urinary 8-isoprostane. CONCLUSION: We found that metabolites in phospholipid groups had strong associations with multiple inflammatory biomarkers, especially CRP, SAA and IL-8. The mechanism of these associations warrants further investigation.

Metabolomic profiling identifies plasma sphingosine 1-phosphate levels associated with welding exposures.

BACKGROUND: Despite a number of known health hazards of welding fume exposure, it is unclear how exposure affects the human metabolome. OBJECTIVE: We assessed the metabolic profiles of welders before and after a 6-hour welding shift, controlling for circadian rhythm of metabolism on a non-welding day. METHODS: Welders were recruited from a training centre in Quincy, Massachusetts, in 2006 and 2010-2012 and donated blood samples on a welding shift day before and after work, as well as on a non-welding day spent in an adjacent classroom. In total, we collected 509 samples from 74 participants. Liquid chromatography-mass spectrometry quantified 665 metabolites from thawed plasmas. Metabolites with significant time (afternoon compared with morning) and day (welding/classroom) interactions were identified by two-way analysis of variance, and the overnight changes were evaluated. RESULTS: Sphingosine 1-phosphate (S1P) and sphingasine 1-phosphate (SA1P) exhibited significant interaction effects between day and time with false discovery rate-adjusted p values of 0.03 and <0.01, respectively. S1P, SA1P and sphingosine shared similar trends over time: high relative levels in the morning of a non-welding day declining by afternoon, but with lower starting levels on a welding day and no decline. There was no obvious pattern related to current smoking status. CONCLUSION: S1P and SA1P profiles were different between welding day and classroom day. The S1P pathway was disrupted on the day of welding exposure. The levels of S1P, SA1P and sphingosine were highly correlated over time. S1P is a signalling lipid with many vital roles; thus, the underlying mechanism and clinical implications of this alteration need further investigation.

Quantitative methods for metabolomic analyses evaluated in the Children's Health Exposure Analysis Resource (CHEAR).

With advances in technologies that facilitate metabolome-wide analyses, the incorporation of metabolomics in the pursuit of biomarkers of exposure and effect is rapidly evolving in population health studies. However, many analytic approaches are limited in their capacity to address high-dimensional metabolomics data within an epidemiologic framework, including the highly collinear nature of the metabolites and consideration of confounding variables. In this Children's Health Exposure Analysis Resource (CHEAR) network study, we showcase various analytic approaches that are established as well as novel in the field of metabolomics, including univariate single metabolite models, least absolute shrinkage and selection operator (LASSO), random forest, weighted quantile sum (WQSRS) regression, exploratory factor analysis (EFA), and latent class analysis (LCA). Here, in a Bangladeshi birth cohort (n = 199), we illustrate research questions that can be addressed by each analytic method in the assessment of associations between cord blood metabolites (1H NMR measurements) and birth anthropometric measurements (birth weight and head circumference).

Determinants of arsenic methylation efficiency and urinary arsenic level in pregnant women in Bangladesh.

BACKGROUND: Prenatal inorganic arsenic (iAs) exposure is associated with pregnancy outcomes. Maternal capabilities of arsenic biotransformation and elimination may influence the susceptibility of arsenic toxicity. Therefore, we examined the determinants of arsenic metabolism of pregnant women in Bangladesh who are exposed to high levels of arsenic. METHODS: In a prospective birth cohort, we followed 1613 pregnant women in Bangladesh and collected urine samples at two prenatal visits: one at 4-16 weeks, and the second at 21-37 weeks of pregnancy. We measured major arsenic species in urine, including iAs (iAs%) and methylated forms. The proportions of each species over the sum of all arsenic species were used as biomarkers of arsenic methylation efficiency. We examined the difference in arsenic methylation using a paired t-test between first and second visits. Using linear regression, we examined determinants of arsenic metabolism, including age, BMI at enrollment, education, financial provider income, arsenic exposure level, and dietary folate and protein intake, adjusted for daily energy intake. RESULTS: Comparing visit 2 to visit 1, iAs% decreased 1.1% (p <  0.01), and creatinine-adjusted urinary arsenic level (U-As) increased 21% (95% CI: 15, 26%; p <  0.01). Drinking water arsenic concentration was positively associated with iAs% at both visits. When restricted to participants with higher adjusted urinary arsenic levels (adjusted U-As > 50 µg/g-creatinine) gestational age at measurement was strongly associated with DMA% (ß = 0.38, p <  0.01) only at visit 1. Additionally, DMA% was negatively associated with daily protein intake (ß = - 0.02, p <  0.01) at visit 1, adjusting for total energy intake and other covariates. CONCLUSIONS: Our findings indicate that arsenic metabolism and adjusted U-As level increase during pregnancy. We have identified determinants of arsenic methylation efficiency at visit 1.

Gene-environment interaction and maternal arsenic methylation efficiency during pregnancy.

BACKGROUND: Single nucleotide polymorphisms (SNPs) may influence arsenic methylation efficiency, affecting arsenic metabolism. Whether gene-environment interactions affect arsenic metabolism during pregnancy remains unclear, which may have implications for pregnancy outcomes. OBJECTIVE: We aimed to investigate main effects as well as potential SNP-arsenic interactions on arsenic methylation efficiency in pregnant women. METHOD: We recruited 1613 pregnant women in Bangladesh, and collected two urine samples from each participant, one at 4-16 weeks, and the second at 21-37 weeks of pregnancy. We determined the proportions of each arsenic metabolite [inorganic As (iAs)%, monomethylarsonic acid (MMA)%, and dimethylarsinic acid (DMA)%] from the total urinary arsenic level of each sample. A panel of 63 candidate SNPs was selected for genotyping based on their reported associations with arsenic metabolism (including in As3MT, N6AMT1, and GSTO2 genes). We used linear regression models to assess the association between each SNP and DMA% with an additive allelic assumption, as well as SNP-arsenic interaction on DMA%. These analyses were performed separately for two urine collection time-points to capture differences in susceptibility to arsenic toxicity. RESULT: Intron variants for As3MT were associated with DMA%. rs9527 (ß = -2.98%, PFDR = 0.008) and rs1046778 (ß = 1.64%, PFDR = 0.008) were associated with this measure in the early gestational period; rs3740393 (ß = 2.54%, PFDR = 0.002) and rs1046778 (ß = 1.97%, PFDR = 0.003) in the mid-to-late gestational period. Further, As3MT, GSTO2, and N6AMT1 polymorphisms showed different effect sizes on DMA% conditional on arsenic exposure levels. However, SNP-arsenic interactions were not statistically significant after adjusting for false discovery rate (FDR). rs1048546 in N6AMT1 had the highest significance level in the SNP-arsenic interaction test during mid-to-late gestation (ß = -1.8% vs. 1.4%, PGxE_FDR = 0.075). Finally, As3MT and As3MT/CNNM2 haplotypes were associated with DMA% at both time points. CONCLUSION: We found that not all genetic associations reported in arsenic methylation efficiency replicate in pregnant women. Arsenic exposure level has a limited effect in modifying the association between genetic variation and arsenic methylation efficiency.

Regulation of birthweight by placenta-derived miRNAs: evidence from an arsenic-exposed birth cohort in Bangladesh.

Altered expression of microRNAs (miRNAs) is implicated in fetal growth. However, the mechanisms by which placenta-derived miRNAs regulate birthweight are not well understood. In Phase 1, we compared the expression of 754 miRNAs in the placenta of mothers from two extreme birthweight groups (0.8-2.2 kg vs. 3.3-3.9 kg, n = 77 each) selected from an arsenic-exposed Bangladeshi birth cohort (n = 1,141). We identified 49 miRNAs associated with the extreme birthweight groups and/or gestational age in Phase 1, which were further analyzed in Phase 2 among 364 randomly selected mother-infant pairs. Gestational age was determined by ultrasound. Causal mediation analysis was used to estimate the effect of miRNAs on birthweight considering gestational age a mediator, adjusting for core blood arsenic and other risk factors. miR-1290, miR-195, and let-7g showed significant inverse associations with gestational age, while miR-328 showed significant positive association [false discovery rate (FDR) <0.05]. Via changing gestational age, miR-1290, miR-195, and miR-27a showed significant inverse associations with birthweight, while miR-328 and miR-324-5p showed significant positive associations (FDR <0.05). The effect of miRNAs on birthweight varied by gestational age (for miR-1290, miR-195, miR-328) and in utero arsenic exposure (for miR-1290): stronger effect was observed among infants delivered early in gestation or exposed to higher concentrations of arsenic in cord blood. Gene enrichment analysis with in silico predicted targets identified cell proliferation, inflammation, apoptosis, insulin, and IGF family signaling cascades associated with these miRNAs. Future studies are warranted to replicate these findings and assess these miRNAs as early biomarkers of fetal growth.

A comparison of bone mineral densities and body composition between Southeast Asia college students and Chinese college students.

The aim of this study was to compare bone mineral densities (BMDs) and body composition between Southeast Asia college students and Chinese college students, in order to provide a certain reference enhancing college students' physical fitness.A total of 1694 Chinese college students (294 men and 1400 women, aged 18-22 years) and 250 Southeast Asia college students (148 men and 102 women, aged 19-22 years) were included in the study. Weight, height, and body mass index were measured anthropometrically. BMD values were determined by ultrasound bone densitometer and body composition was determined by body composition analyzer.Southeast Asia college students were overweight than Chinese college students (250 vs 1694) (P < 0.05). Chinese college students had a significantly lower body weight, fat mass, lean tissue mass, lean body weight, estimation of bone mass, protein, and metabolic rate but higher BMD at the calcaneus compared with Southeast Asia college students (P < 0.05 for all parameters). However, body water, intracellular fluid, and extracellular fluid were not significantly different between Chinese college students and Southeast Asia college students (P > 0.01 for all parameters).The results of this cross-sectional study suggest that Chinese college students had a higher BMD but lower body composition than Southeast Asia college students, which may be associated with genes, diet, exercise, and other factors.

Stringent homology-based prediction of H. sapiens-M. tuberculosis H37Rv protein-protein interactions.

BACKGROUND: H. sapiens-M. tuberculosis H37Rv protein-protein interaction (PPI) data are essential for understanding the infection mechanism of the formidable pathogen M. tuberculosis H37Rv. Computational prediction is an important strategy to fill the gap in experimental H. sapiens-M. tuberculosis H37Rv PPI data. Homology-based prediction is frequently used in predicting both intra-species and inter-species PPIs. However, some limitations are not properly resolved in several published works that predict eukaryote-prokaryote inter-species PPIs using intra-species template PPIs. RESULTS: We develop a stringent homology-based prediction approach by taking into account (i) differences between eukaryotic and prokaryotic proteins and (ii) differences between inter-species and intra-species PPI interfaces. We compare our stringent homology-based approach to a conventional homology-based approach for predicting host-pathogen PPIs, based on cellular compartment distribution analysis, disease gene list enrichment analysis, pathway enrichment analysis and functional category enrichment analysis. These analyses support the validity of our prediction result, and clearly show that our approach has better performance in predicting H. sapiens-M. tuberculosis H37Rv PPIs. Using our stringent homology-based approach, we have predicted a set of highly plausible H. sapiens-M. tuberculosis H37Rv PPIs which might be useful for many of related studies. Based on our analysis of the H. sapiens-M. tuberculosis H37Rv PPI network predicted by our stringent homology-based approach, we have discovered several interesting properties which are reported here for the first time. We find that both host proteins and pathogen proteins involved in the host-pathogen PPIs tend to be hubs in their own intra-species PPI network. Also, both host and pathogen proteins involved in host-pathogen PPIs tend to have longer primary sequence, tend to have more domains, tend to be more hydrophilic, etc. And the protein domains from both host and pathogen proteins involved in host-pathogen PPIs tend to have lower charge, and tend to be more hydrophilic. CONCLUSIONS: Our stringent homology-based prediction approach provides a better strategy in predicting PPIs between eukaryotic hosts and prokaryotic pathogens than a conventional homology-based approach. The properties we have observed from the predicted H. sapiens-M. tuberculosis H37Rv PPI network are useful for understanding inter-species host-pathogen PPI networks and provide novel insights for host-pathogen interaction studies.

Stringent DDI-based prediction of H. sapiens-M. tuberculosis H37Rv protein-protein interactions.

BACKGROUND: H. sapiens-M. tuberculosis H37Rv protein-protein interaction (PPI) data are very important information to illuminate the infection mechanism of M. tuberculosis H37Rv. But current H. sapiens-M. tuberculosis H37Rv PPI data are very scarce. This seriously limits the study of the interaction between this important pathogen and its host H. sapiens. Computational prediction of H. sapiens-M. tuberculosis H37Rv PPIs is an important strategy to fill in the gap. Domain-domain interaction (DDI) based prediction is one of the frequently used computational approaches in predicting both intra-species and inter-species PPIs. However, the performance of DDI-based host-pathogen PPI prediction has been rather limited. RESULTS: We develop a stringent DDI-based prediction approach with emphasis on (i) differences between the specific domain sequences on annotated regions of proteins under the same domain ID and (ii) calculation of the interaction strength of predicted PPIs based on the interacting residues in their interaction interfaces. We compare our stringent DDI-based approach to a conventional DDI-based approach for predicting PPIs based on gold standard intra-species PPIs and coherent informative Gene Ontology terms assessment. The assessment results show that our stringent DDI-based approach achieves much better performance in predicting PPIs than the conventional approach. Using our stringent DDI-based approach, we have predicted a small set of reliable H. sapiens-M. tuberculosis H37Rv PPIs which could be very useful for a variety of related studies. We also analyze the H. sapiens-M. tuberculosis H37Rv PPIs predicted by our stringent DDI-based approach using cellular compartment distribution analysis, functional category enrichment analysis and pathway enrichment analysis. The analyses support the validity of our prediction result. Also, based on an analysis of the H. sapiens-M. tuberculosis H37Rv PPI network predicted by our stringent DDI-based approach, we have discovered some important properties of domains involved in host-pathogen PPIs. We find that both host and pathogen proteins involved in host-pathogen PPIs tend to have more domains than proteins involved in intra-species PPIs, and these domains have more interaction partners than domains on proteins involved in intra-species PPI. CONCLUSIONS: The stringent DDI-based prediction approach reported in this work provides a stringent strategy for predicting host-pathogen PPIs. It also performs better than a conventional DDI-based approach in predicting PPIs. We have predicted a small set of accurate H. sapiens-M. tuberculosis H37Rv PPIs which could be very useful for a variety of related studies.

[The clinical study on the surgical treatment of thoracic aortic aneurysm associated with coronary artery disease].

OBJECTIVE: To analyze the factors which influence the safety and prognosis of aorta replacement combined with coronary artery bypass grafting (CABG) for thoracic aortic aneurysm associated with coronary artery disease. METHODS: From May 1982 to October 2002, 67 patients with thoracic aortic aneurysm were admitted, and 24 of them combined with CABG. Of the 24 patients, 9 received descending aorta replacement combined with CABG, and the other 15 received the ascending aorta replacement combined with CABG. The treatment results were compared with the other 43 patients only undergoing the thoracic aortic replacement. RESULTS: The mortality rate of the patients with aorta replacement combined with CABG was 13% (3/24). Though the descending aorta replacement combined with CABG could make the cardiopulmonary bypass time and selective cerebral perfusion time longer, (278 +/- 54) min and (188 +/- 59) min respectively, no significant difference was observed in postoperative complications, 3-year survival rate, 3-year-cardiac-event-free rate compared with the patients only undergoing the thoracic aortic replacement (P > 0.05). CONCLUSIONS: The aorta replacement combined with CABG can be performed safely, and the revascularization for coronary artery disease is useful for preventing occurrence of cardiac events.

Effect of four crystalloid cardioplegias on immature rabbit hearts during global ischaemia.

OBJECTIVE: Controversy surrounds the reported beneficial effects of crystalloid cardioplegic solutions in the immature myocardium. In the present study, we investigated the efficacy of four clinical cardioplegic solutions in the immature myocardium to determine if cardioplegic protection could be demonstrated and, if yes, the relative efficacy of the four solutions. METHODS: Isolated, working hearts (n=6 per group) from neonatal rabbits (age, 7-14 days) were perfused aerobically (37 C) for 15 minutes in the Langendorff mode and 30 minutes in the working mode before a 2-minute infusion of one of four cardioplegic solutions: the modified St. Thomas' Hospital no. 1 cardioplegic solution, Tyers solution, Bretschneider solution or Roe solution. Hearts were then rendered globally ischaemic for 120 minutes at 14C before reperfusion for 15 minutes in the Langendorff mode and 30 minutes in the working mode. The post-ischaemic recovery of cardiac function and leakage of myocardial enzymes (GOT, CK, CK-MB, LDH, LDH1) were compared with results in non-cardioplegic control hearts. RESULTS: Good protection was observed with modified St. Thomas' Hospital and Tyers solutions: postischaemic recovery of cardiac output was increased from 80.43+/-3.62% in the non-cardioplegic group to 85.19+/-3.12% and 70.66+/-3.48% in the St. Thomas' Hospital and Tyers groups (p<0.05), respectively. In contrast, no obvious protection was observed with either the Bretschneider or Roe solutions: cardiac output recovered to 45.08+/-3.16% and 30.06+/-2.59%, respectively. Post-ischaemic CK leakage was 19.83+/-2.14 IU/mL and 21.17+/-2.32 IU/mL in the St. Thomas' Hospital and Tyers groups (p>0.05). In the Bretschneider group, CK leakage increased to 30.00+/-3.16 IU/mL (p<0.01 vs. non-cardioplegic control hearts), and in the Roe group, CK leakage was 31.00+/-5.10 IU/mL (p<0.05 vs. cardioplegic-free hearts). Post-ischaemic ATP was 1.98+/-0.54 micromol/g*dry weight and 1.35+/-0.39 micromol/g*dry weight in the St. Thomas' Hospital and Tyers groups (p<0.01 vs. non-cardioplegic control group), respectively. In the Bretschneider group, ATP decreased to 0.91+/-0.16 micromol/g*dry weight (p<0.05 vs. non-cardioplegic control hearts), and in the Roe group to 0.88+/-0.10 micromol/g*dry weight (p<0.01 vs.cardioplegic-free hearts). CONCLUSION: In conclusion, cardioplegic protection can be achieved in the immature rabbit myocardium with both St. Thomas' Hospital and Tyers solutions, but acalcaemic solutions such as Bretschneider and Roe solutions increased damage. The reported lack of cardioplegic efficacy in the immature myocardium may, therefore, reflect the choice of cardioplegic solution rather than a greater vulnerability to injury in the neonatal heart.

Surgical treatment for giant solid tumors of the mediastinum: a study of 26 cases.

Twenty-six patients with giant solid tumors of the mediastinum (GSTM) were treated surgically from 1975 to 2000. Femorofemoral cardiopulmonary bypass (CPB) was used before induction of anesthesia as a precaution against total tracheal occlusion in two cases. Resection of the tumor was accomplished in all patients, combined with partial pericardium resection in five cases, left upper lobe of lung resection in two cases, and reconstruction of the superior vena cava (SVC) and innominate vein in four cases. The weights of resected tumors ranged from 1.1 to 4.8 kg, with an average of 2.2 kg. The majority were benign (22 of 26, 84.6%). The postoperative complications included two cases with recurrent laryngeal nerve injury, three cases with wound infection, and two cases with dilatant pneumonedema. Diagnosis of GSTM was not difficult based on imaging and needle biopsy. Femorofemoral CPB is recommended before induction of anesthesia for patients with superior airway obstruction and superior vena cava occlusion. Perioperative management includes strict hemostasis, proper chest wall reconstruction, and prevention of re-expansion pulmonary edema.