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Context Ambrisentan is an oral endothelin-receptor antagonist (ERA). However, there is no report on the interaction between ambrisentan and shikonin. Objective To investigate the effect of shikonin on ambrisentan metabolism in vivo and in vitro. Materials and methods This study developed an ultra-performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) method for simultaneous determination of ambrisentan and (S)-4-hydroxymethyl ambrisentan in rat plasma. Twelve male Sprague-Dawley (SD) rats were divided into two groups (n = 6): the control group and shikonin (20 mg/kg) group. The pharmacokinetics of ambrisentan (2.5 mg/kg) were investigated after 30 min. Additionally, human and rat liver microsomes were used to investigate the herb-drug interaction. Results The UPLC-MS/MS method was shown to be accurate, precise and reliable, and was successfully applied to the herb-drug interaction study of ambrisentan with shikonin. When co-administrated with 20 mg/kg shikonin, the Cmax and AUC(0–∞) of ambrisentan were significantly increased by 44.96 and 16.65%, respectively (p

Rice blast (the causative agent the fungus Magnaporthe oryzae) represents a major constraint on the productivity of one of the world’s most important staple food crops. Genes encoding resistance have been identified in both the Xian and Geng subspecies genepools, and combining these within new cultivars represents a rational means of combating the pathogen. In this research, deeper allele mining was carried out on Pid2, Pid3, and Pid4 via each comprehensive FNP marker set in three panels consisting of 70 Xian and 58 Geng cultivars. Within Pid2, three functional and one non-functional alleles were identified; the former were only identified in Xian type entries. At Pid3, four functional and one non-functional alleles were identified; once again, all of the former were present in Xian type entries. However, the pattern of variation at Pid4 was rather different: here, the five functional alleles uncovered were dispersed across the Geng type germplasm. Among all the twelve candidate functional alleles, both Pid2-ZS and Pid3-ZS were predominant. Furthermore, the resistance functions of both Pid2-ZS and Pid3-ZS were assured by transformation test. Profiting from the merits of three comprehensive FNP marker sets, the study has validated all three members of the Pid family as having been strictly diverged into Xian and Geng subspecies: Pid2 and Pid3 were defined as Xian type resistance genes, and Pid4 as Geng type. Rather limited genotypes of the Pid family have been effective in both Xian and Geng rice groups, of which Pid2-ZS_Pid3-ZS has been central to the Chinese rice population.

Rice blast (the causative agent the fungus

Background: Rice blast (causative agent the fungus Pyricularia oryzae) represents a major constraint over the productivity of one of the world’s most important staple foods. Genes encoding resistance have been identified in both the indica and japonica subspecies genepools and combining these within new cultivars represents a rational means of combating the pathogen.Results: In this research, a deeper allele mining was carried out on Pid-2, Pid-3, and Pid-4 by each of comprehensive FNP marker set in the three panels consisting of 70 indica and 58 japonica cultivars. Within Pid-2, three functional and one non-functional alleles were identified; the former were only identified in indica type entries. At Pid-3, four functional and one non-functional alleles were identified, and once again, all of the formers were present in indica type entries. However, the pattern of variation at Pid-4 was rather different: here, the five functional alleles uncovered were dispersed across the japonica type germplasm. Among all the 12 candidate functional alleles, both Pid2-ZS and Pid3-ZS were predominant. Furthermore, the resistance functions of both Pid2-ZS and Pid3-ZS were assured by transformation test. Conclusions: Profiting from merits of three comprehensive FNP marker sets, the study has validated that all three members of the Pid family have been strictly diverged into indica and japonica subspecies: Pid-2 and Pid-3 were defined as indica type resistance genes, and Pid-4 as japonica one. Rather limited genotypes of the Pid family have been effective in both indica and japonica rice groups, of which Pid2-ZS+Pid3-ZS has been central to the Chinese rice population.

Background Millions of adults have been reported with hyperlipemia in the world. It is still unclear whether the plasma level of essential amino acids (AAs) will be influenced by the hyperlipemia. This study was aimed to investigate the AAs levels and the underlying metabolic relationship in hyperlipidemic subjects. Methods An ultra-high performance liquid chromatography-tandem mass spectrometric (UPLC-MS/MS) method was developed for the determination of phenylalanine (Phe), valine (Val), histidine (His), tryptophan (Trp), and methionine (Met). Plasma samples (100 μL) were precipitated by acetonitrile (300 μL) and analyzed on a BEH C18 (2.1 mm × 100 mm, 1.7 μm) column at 40 °C by gradient elution. The mobile phase composed of 0.1% formic acid and acetonitrile was used with flow rate at 0.2–0.4 ml/0-3 min. Five AAs were determined at positive electrospray ionization (ESI+) at m/z 118.1/72.1 (Val), 150.12/104.02(Met), 156.06/110.05(His), 166.1/120.1(Phe), and 205.2/188.02 (Trp). A total of 75 healthy subjects and 83 hyperlipidemic subjects, who had blood routine test and plasma lipid test were determined by developed UPLC-MS/MS. Results It was shown that there was good linearity for Val, Met, His, Phe, and Trp within 1–100 μg/mL. The relative standard deviations of precision and accuracy were all within 15%. The level of Val, Phe, Trp, His, and Met were 35.34 ± 15.64, 22.72 ± 9.13, 17.23 ± 4.94, 16.78 ± 13.64, and 6.24 ± 1.97 μg/mL in healthy subjects, while they were 38.04 ± 16.70, 22.41 ± 8.45, 15.62 ± 5.77, 18.35 ± 14.49, and 6.21 ± 1.97 μg/mL in hyperlipidemic subjects respectively. The Spearman’s correlations analysis showed that there were high correlations between Val, Phe, Trp, His, Met and triglyceride in healthy subjects. While, those correlations decreased in hyperlipemia cases. Conclusion A convenient and sensitive method for simultaneous determination of Val, Phe, Trp, His, and Met in human plasma was developed. There was a high correlation between Val, Phe, Trp, His, Met and triglyceride. Hyperlipemia influences the metabolic balance of His, Phe, Trp, Met and Val.

Multiple myeloma (MM) is a pernicious plasma cell disorder and has a poor prognosis. N

Objective To analyse the clinical characteristics of extra-thyroid 99mTc-pertechnetate uptake in order to explore the effect of the phenomenon on radioactive iodine (RAI) therapy for differentiated thyroid carcinoma (DTC) and its clinical significance. Methods This study retrospectively selected patients with DTC and extra-thyroid 99mTc-pertechnetate uptake. The clinical features, location, location count and extra-thyroid 99mTc-pertechnetate uptake distribution were analysed, combined with the uptake rate, stimulated thyroglobulin (sTg) level, post-therapy whole-body scan and curative effect. Results A total of 38 patients were enrolled in the study and 65 extra-thyroid 99mTc-pertechnetate foci were detected. Thirty-four patients showed abnormal 99mTc-pertechnetate uptake in the lymph nodes (26 of 38; 68.4%), lungs (four of 38; 10.5%) and bones (four of 38; 10.5%). The corresponding uptake rates were 0.2%, 0.2% and 0.8%, respectively. The uptake rate and sTg were significantly positively correlated ( r = 0.36). 131I uptake was found in 36 patients at the 99mTc-pertechnetate uptake site. The number of iodine uptake foci was significantly higher than that of 99mTc-pertechnetate uptake foci. The sTg value and pathological staging significantly differed between the excellent and nonexcellent response groups (Z = –2.947 and Z = –2.348, respectively). Conclusion Extra-thyroid 99mTc-pertechnetate uptake mostly indicated metastases with specific clinical features, which may have prognostic value for the judgment of iodine uptake function and the RAI therapy plan.

The expression levels of microRNA (miR)-221-3p and miR-222-3p in thyroid cancer have been found to be upregulated compared with those in normal tissues. The present study aimed to determine the effects and potential underlying mechanisms of miR-221-3p and miR-222-3p on the regulation of radioactive iodine (131I) uptake and radiosensitivity of thyroid cancer cells. The potential regulatory target genes of miR-221-3p and miR-222-3p were predicted by bioinformatics analysis, and reverse transcription-quantitative polymerase chain reaction was used to verify miR-221-3p, miR-222-3p and target gene expression levels in thyroid cancer tissues and cell lines. Overexpression of miR-221-3p or miR-222-3p in cell models was performed using lentivirus infection. Knockdown of miR-221-3p and miR-222-3p in cells was achieved using oligonucleotide inhibitor transfection. Western blotting was used to analyze the expression levels of target proteins. In addition, the effects of miR-221-3p and miR-222-3p on the radiosensitivity of thyroid cancer cells were verified using a colony formation assay. The results of the present study revealed that the expression levels of miR-221-3p and miR-222-3p were significantly upregulated, while the expression levels of suppressor of cytokine signaling 3 (SOCS3) were downregulated in thyroid cancer tissues. Furthermore, miR-221-3p and miR-222-3p overexpression downregulated the expression levels of SOCS3, E-cadherin and solute carrier family 5 member 5 (NIS), and upregulated the expression levels of phosphorylated STAT3 and vimentin. Following the overexpression of miR-221-3p or miR-222-3p in the FTC133 and TPC1 cell lines, their radiosensitivity was suppressed. In conclusion, the findings of the present study suggested that miR-221-3p and miR-222-3p may downregulate the expression levels of NIS and promote radioresistance. The potential mechanism was hypothesized to be associated with the miR-221-3p and miR-222-3p targeting of the SOCS3 gene, which may subsequently activate the STAT3 signaling pathway.

Background: Over the last few decades, extra-thyroid 99mTc-pertechnetate uptake has rarely been reported. The clinical characteristics of extra-thyroid 99mTc-pertechnetate uptake were retrospectively analysed to explore the effect of the phenomenon on RAI therapy for DTC and its clinical significance. Methods: In this study, we retrospectively analysed 4930 RAI-treated DTC patients who had undergone 99mTc-pertechnetate scanning. Thirty-eight cases with extra-thyroid 99mTc-pertechnetate uptake were selected. The clinical features, location, location count and extra-thyroid 99mTc-pertechnetate uptake distribution were analysed, combined with the uptake rate, stimulated thyroglobulin (sTg) level, post-therapy whole-body scan and curative effect. Results: The results showed that sixty-five extra-thyroid 99mTc-pertechnetate foci were detected in 38 patients. The proportions of patients with abnormal uptake in the lymph nodes, lungs and bones were 68.4%, 10.5% and 10.5%, respectively. The corresponding uptake rates were 0.2%, 0.2% and 0.8%. The uptake rate was significantly lower in the lymph nodes than in the bones (Z = -2.722, p = 0.019). The uptake rate and sTg were positively correlated (r = 0.36, p = 0.027). 131I uptake was found in 36 cases at the technetium uptake site, and the number of iodine uptake foci was significantly higher than that of 99mTc-pertechnetate uptake foci. The sTg value and pathological staging significantly differed between the excellent and nonexcellent response groups (Z = 2.947, p = 0.003 and Z = 2.348, p = 0.019, respectively). Conclude: Extra-thyroid 99mTc-pertechnetate uptake mostly indicated metastases with specific clinical features, which may have prognostic value for the judgment of iodine uptake function and the RAI therapy plan.

Ivosidenib, as an oral mutant isocitrate dehydrogenase 1 (mIDH1) inhibitor, was awarded approval in the USA for the targeted therapy of relapsed or refractory acute myeloid leukemia (AML) in adult patients, who also had a susceptible enzyme to mIDH1. The aim of our present study was to develop and validate an accurate and fast assay based on the ultrahigh-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) technique for the quantification of ivosidenib in plasma and to investigate the possible effects of different CYP3A4 inhibitors (voriconazole, itraconazole and fluconazole) on ivosidenib metabolism in rats. After the fast protein crash with acetonitrile, chromatographic separation of ivosidenib and erlotinib (used as the internal standard in this experiment, IS) was accomplished using an Acquity BEH C18 (2.1 mm × 50 mm, 1.7 μm) column, and detection of the analyte was also performed using a Xevo TQ-S triple quadrupole tandem mass spectrometer in the positive ion electrospray ionization (ESI) interface. The assay showed enough linearity over a 0.5−6000 ng/mL calibration range. The application of the validated bioanalytical method based on the UHPLC-MS/MS technique was further successfully exhibited in an animal study of the drug-drug interaction between ivosidenib (50 mg/kg) and voriconazole (20 mg/kg)/itraconazole (20 mg/kg)/fluconazole (20 mg/kg) in rats. Voriconazole, itraconazole and fluconazole increased the exposure of ivosidenib in plasma by different degrees and also had a potential inhibitory effect on the metabolism of ivosidenib. Thus, a dose reduction or interruption of ivosidenib may be important to guide the practice of clinical medicine.

Additional file 1 Supplement Table 1 The equation of calibration curves of 5 AAs in human plasma

Additional file 5 Supplement Table 5 Peaks area of 5 AAs (10 μg/mL) in pure standard and blood sample determined by Hilic and BEH C18 column

Additional file 4 Supplement Table 4 RSD values of ruggedness determined on two different columns and by different analysts

Additional file 2 Supplement Table 2 The extraction recovery, matrix effect and stability of 5 AA in human plasma (n = 3)

Abstract: Objective To investigate the effect of pomalidomide on the maturation of monocyte derived dendritic cells (moDCs) from healthy donors (HDs) and multiple myeloma (MM) patients. Method Peripheral blood mononuclear cells (PBMCs) were isolated from the peripheral blood of HDs and MM patients. The generation of moDCs from monocytes in PBMCs was conducted by the incubation of 7 days in a medium consisting of RPMI 1640 medium, 5% human serum, 800U/mL GM-CSF, 500U/mL IL-4, 100U/mL penicillin and 0.1mg/mL streptomycin. During this period, the incubation system was administrated with pomalidomide at 10 µM or 1×PBS as the control group. On the 8 th day, cells were harvested, and the immunophenotyping of cells were analyzed by the flow cytometry. The CD80+CD86+ cell population in total cells is gated as DCs in the FACS analyzing system. Then, the median fluorescence intensity (MFI) of surface markers CD40 and HLA-DR as well as the proportion of CD40+ DCs and HLA-DR+ DCs in total DCs were analyzed respectively. In addition, supernatant from the incubation system with or without pomalidomide administration was collected and detected for the concentration of cytokines IL-12, TNF-α and MIP-1α. Results The proportion of CD80+CD86+ cells in total cells was higher in HD group (n=15) than in MM patient group (n=11), but there was no statistically significant difference (93.49%±6.43% vs 77.04%±27.17%, P=0.094). When analyzing all the HD-derived moDCs (n=15), pomalidomide significantly enhanced the MFI of CD40 (P=0.003) and HLA-DR (P=0.040) on moDCs when compared with the control group. Meanwhile, the proportion of CD40+ DCs (P=0.008) and HLA-DR+ DCs (P=0.032) in total DCs was significantly higher in pomalidomide group than in control group. When analyzing all MM patients-derived moDCs (n=11), pomalidomide significantly enhanced the MFI of CD40 (P=0.047) and HLA-DR (P=0.006) on moDCs when compared with the control group. Meanwhile, the proportion of HLA-DR+ DCs in total DCs was significantly higher in pomalidomide group than in the control group (P=0.000). Moreover, pomalidomide treated HD-derived moDCs (n=8) produced 192% IL-12 (P=0.020), 110% TNF-α (P=0.006) and 112% MIP-1α (P=0.055) of untreated moDCs. However, when analyzing MM patients-derived moDCs (n=10), the expression of IL-12 (P=0.458), TNF-α (P=0.377) and MIP-1α (P=0.248) from moDCs showed no significant difference between pomalidomide group and the control group. Conclusions Pomalidomide at 10 µM can promote the maturation of both HD-derived moDCs and MM patients-derived moDCs. Pomalidomide shows potential to be a DC adjuvant for DC-based therapeutic strategies, such as DC vaccine and DC cell-therapy in MM. Key words: Pomalidomide; Monocyte derived Dendritic Cells; Multiple Myeloma; DC Adjuvant Disclosures No relevant conflicts of interest to declare.

Backgroud: Reactivation of nasopharyngeal carcinoma virus (EBV) post allogeneic hematopoietic stem cell transplantation(allo-HSCT) is very common, but sustained EBV activation maybe induce EBV-associated lymphoproliferative disease (PTLD), transplant-associated hemophagocytic syndrome and thrombocytopenia after transplantation, which is extremely harmful for long-term survival. Currently, the first-line therapeutic strategy for sustained EBV activation is rituximab, but rituximab leads to increased rates of infections and delayed immune reconstitution. Therefore, for the patients with EBV-DNA copies under 1000000/mL, the pros and cons of rituximab are worth weighing. Objective: To evaluate the efficacy and safety of high-dose intravenous acyclovir in the treatment of EBV reactivation post allo-HSCT. Method: Retrospective analysis of clinical data of patients with EBV reactivation post transplantation treated with high-dose intravenous acyclovir (0.5g Q8H) in Sichuan Provincial People's Hospital from January 2019 to May 2021. A total of 49 patients post allo-HSCT from from January 2019 to May 2021 were enrolled in this study. In this population, 38 patients accepted haplo-SCT and 11 patients accepted sibling-SCT. These patients all don't suffer from PTLD and transplant-associated hemophagocytic syndrome, but are resistant to oral antiviral drugs (Acyclovir, valacyclovir, and famciclovir). Simultaneously with intravenous acyclovir, all patients received human immunoglobulin with one dose of 0.4g/kg. The clinical efficacy was evaluated as follows: Overall response(ORR) was defined as the decrease of EBV copies; Complete remission(CR) was defined as the negative turn of EBV copies; Partial response(PR), Stable disease(SD) and Progression of disease(PD) were respectively defined as the decrease, no significant change and increase of EBV copies. Results: The median transplantation time was +86 (+38 to +1587) days. The median number of EBV-DNA copies/ml was 177000 (6620-8200000). After treatment with high-dose intravenous acyclovir, 29 (59.2%) of 49 patients achieved CR, 10 (20.4%) of 49 patients achieved PR, and 10 (20.4%) of 49 patients had no response, including 2 patients with SD and 8 patients with PD. All 49 patients had responded to this regimen with 79.6% ORR. The median time of response and negative turn was 4 days (2 to 11 days) and 9 days (2 to 23 days) respectively. After 100 days for follow-up, 14 of 39 responding patients suffered from EBV activation again, and the EBV copies were higher than that when high-dose acyclovir was initiated. All 24 patients who did not turn negative were recommended with the treatment of rituximab, and 19 of 20 patients who completed treatment turned negative during the three months post acyclovir treatment. The main adverse events (AEs) of this regimen were neutropenia (CTCAE grade 2-3, 8 in 49 patients), thrombocytopenia (CTCAE grade 2-3, 10 in 49 patients), increased serum creatinine (CTCAE grade 1-2, 4 in 49 patients), phlebitis (6 in 49 patients). These AEs were all reversible, which recovered after withdrawal. There was no significant change in the counts of CD4 + T cells and CD8 + T cells before and after intravenous high-dose acyclovir. Conclusion: High-dose intravenous acyclovir is safe and effective in the treatment of EBV reactivation post allo-HSCT, although nearly half of patients still need rituximab treatment. It is a reasonable strategy for patients with oral antiviral resistance. Most patients can avoid the prescription of rituximab in the first six months post allo-HSCT, without interference of immune reconstitution. Disclosures No relevant conflicts of interest to declare.

The purpose of the experiment was mainly to establish and verify a precise and straightforward ultra-performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) approach for simultaneously analyzing the concentration levels of amiodarone, dronedarone, and their metabolites (desethylamiodarone and desbutyldronedarone) in the plasma of Sprague-Dawley (SD) rats and to investigate the pharmacokinetics of all analytes in SD rats. After rapid protein precipitation by using acetonitrile, we accomplished the chromatographic separation of amiodarone, desethylamiodarone, dronedarone, desbutyldronedarone and ivabradine (internal standard, IS) by using Acquity BEH C18 column and detected through a mass spectrometer with Xevo TQ-S triple quadrupole tandem, choosing the positive ion mode. The approach showed wonderful linearity, and the range of calibration curve for amiodarone was 1–200 ng/mL, desethylamiodarone was 0.1–20 ng/mL, dronedarone was 0.5–100 ng/mL, and desbutyldronedarone was 0.25–50 ng/mL, respectively. For lower limit of quantification (LLOQ), the current method of UPLC-MS/MS can achieve values of 1.0 ng/mL for amiodarone, 0.1 ng/mL for desethylamiodarone, 0.5 ng/mL for dronedarone, and 0.25 ng/mL for desbutyldronedarone, respectively. The accuracy of intra-day and inter-day of all analytes was between −14.8% to 10.9%, while the precision was ≤ 13.3%. For each substance, the recovery rate was > 82.1%, besides, obvious matrix effect was not found. In all conditions, the stability of all analytes was comfirmed to the plasma sample quantification. In addition, the method of UPLC-MS/MS we developed could also be applied to measure the pharmacokinetic characteristics including amiodarone, desethylamiodarone, dronedarone, and desbutyldronedarone in the plasma of SD rats.

In this study, low temperature plasma nitriding (LPN) was primarily used in 38CrMoAl hydraulic plunger to overcome the peeling problem of the compound layer in real application. The cross-sectional microstructures of the treated samples were observed by optical metallography, phase constituents were determined by X-ray diffraction (XRD), the microhardness profile and brittleness of surface were measured by microhardness tester. The results showed that no compound layer was formed while plasma nitriding at 450 °Cfor 6 h, and XRD also confirmed that there existed no γ′-Fe 4 N. Meanwhile, the brittleness was decreased at lower temperature plasma nitriding comparing with normal plasma nitriding (NPN), due to no brittle compound layer formed on the outmost layer.

Purpose: The purpose of this study is to examine the effectiveness of introducing both rituximab (RTX) and 131I for active Graves' ophthalmopathy (GO) with hyperthyroidism. Methods: In total, 217 patients suffering from active GO with hyperthyroidism were included in this research. All subjects were randomly assigned to 3 groups. Patients in group A solely received 131I treatment; group B1 underwent a methylprednisolone treatment in combination with 131I treatment; and group B2 received an RTX in combination with 131I treatment. Hyperthyroidism treatment outcomes, orbital volumetry, ophthalmic assessments, serum cytokine levels, and adverse effects were measured after treatment. Results: The orbital volumetry principle was significantly different from 24 weeks after the start of treatment among all 3 groups, and improvements in most ophthalmic parameters were regarded significantly different among 3 groups (all p < 0.05). The expression levels of miR-146a and most serum cytokines were regarded significantly different from 24 weeks after the start of treatment among 3 groups (all p < 0.05). Conclusions: In comparison with other therapies, RTX treatment in combination with 131I treatment is considered to be more effective for hyperthyroidism with active GO.

The overall survival of multiple myeloma (MM) patients significantly improved with the use of proteasome inhibitor such as bortezomib. However, resistance to sorafenib limits its use. Bortezomib-resistant MM cells were generated and their bortezomib-resistant properties were confirmed by cell viability and apoptosis assays. To explore functions and underlying mechanisms of long non-coding RNA nuclear paraspeckle assembly transcript 1 (NEAT1) on bortezomib resistance in MM, MTT assays, flow cytometry analyses, dual luciferase report gene assays, RNA pulldown assays and chromatin immunoprecipitation assays were carried out. NEAT1 and specific protein 1 (Sp1) was upregulated while miR-29b-3p was down regulated in bortezomib-resistant MM cells. NEAT1 promoted Sp1 expression by sponging miR-29b-3p and then enhanced the tolerance of MM cells to bortezomib. Sp1 targeted to NEAT1 promoter region promoting NEAT1 transcription and formed a positive feedback loop. NEAT1 and Sp1 levels were higher and miR-29b-3p was levels were lower in bortezomib-resistant MM patients. NEAT1/miR-29b-3p/Sp1 feedback loop enhanced the tolerance of MM cells to bortezomib. These results indicate potentially valuable targets for overcoming bortezomib resistance for MM.

In the present study, an accurate, simple and fast bioanalytical method based on ultra performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) technique for simultaneous quantification of plasma selexipag and its main metabolite ACT-333679 concentrations in rats was optimized and established. The purpose of chromatographic separation of selexipag, ACT-333679 and the internal standard (IS, diazepam) was accomplished using an Acquity BEH C18 (2.1 mm × 50 mm, 1.7 μm) column. The mobile phase was consisted of acetonitrile (solution A) and 0.1 % formic acid in water (solution B) in a linear gradient elution procedure with a flow rate of 0.40 mL/min. The measurement of the analytes and IS was explored using a XEVO TQ-S triple quadrupole tandem mass spectrometer, which was comprised with electrospray ionization (ESI) source in positive ion mode. Selected multiple reaction monitoring (MRM) mode was employed to detect the parent-to-daughter ion transitions as follows: m/z 497.4 → 302.2 for selexipag, m/z 420.1 → 378.2 for ACT-333679, and m/z 285.0 → 154.0 for diazepam (IS), respectively. The new UPLC-MS/MS method showed good linearity respectively at the calibration curve range of 0.05-50 ng/mL for selexipag, and 0.05-250 ng/mL for ACT-333679. The intra- and inter-day of accuracy and precision were all within the acceptable limits in the bioanalytical method, and the results of recovery and matrix effect were also met the requirements. The newly developed UPLC-MS/MS assay was forward successfully used to describe the pharmacokinetic profiles of selexipag and ACT-333679 in rats after oral treatment with 6.0 mg/kg selexipag.

Eliglustat is an oral substrate reduction therapy drug and has been approved as a first-line treatment for adults with Gaucher disease type 1 (GD 1). In the present study, we aimed to develop and establish an accurate and simple ultra performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) method for the measurement of eliglustat concentration in rat plasma. The goal of chromatographic separation of eliglustat and the internal standard (bosutinib) was finished on an Acquity BEH C18 (2.1 mm × 50 mm, 1.7 μm) column. Acetonitrile and 0.1% formic acid in water were employed as the mobile phase in a mode of gradient elution with the 0.40 mL/min flow rate. The detection was carried out on a XEVO TQ-S triple quadrupole tandem mass spectrometer coupled with electrospray ionization (ESI) interface in the positive-ion mode. Multiple reaction monitoring (MRM) was used to monitor the precursor-to-product ion transitions of m/z 405.4 → 84.1 for eliglustat and m/z 530.2 → 141.2 for bosutinib (IS), respectively. It was found that the linearity of the method in the range of 1–500 ng/mL was good for eliglustat. The values of intra- and inter-day accuracy and precision were all within the acceptance limits, and no matrix effect was found in this method. The current developed method was further performed to support in vivo pharmacokinetic study of eliglustat after oral treatment with 10 mg/kg eliglustat to rats.

Linezolid has been widely used in serious infections for its effective inhibiting effect against multidrug-resistant gram-positive pathogens. However, linezolid caused severe adverse reactions, such as thrombocytopenia, anaemia, optic neuropathy, and near-fatal serotonin syndrome. In order to investigate the toxicity of linezolid, twenty-four Sprague-Dawley rats were randomly divided into: control group (n=7), low-group (n=8), and high-group (n=9). The rats of low-group and high-group were given by gavage with linezolid 60 and 120 mg/kg/day for 7 days, respectively. The serum concentration of linezolid was determined by high performance liquid chromatography (HPLC); blood metabolic change was analyzed by gas chromatography-mass spectrometer (GC-MS). Adenosine triphosphate (ATP) concentration in HepG2-C3A after being cultured with linezolid was determined by HPLC. The results showed that there were six metabolites and nine metabolites had statistical differences in low-group and high-group (Pα-linolenic acid, eicosapentaenoic acid, 9,12-Octadecadienoic acid, and docosahexaenoic acid were all decreased in low-group and high-group. ATP concentration was decreased in HepG2-C3A after cultured with linezolid. In conclusion, the toxicity of linezolid is related to its serum concentration. Linezolid may inhibit the synthesis of ATP and fatty acid.

Plasma post-oxidizing was conducted immediately after plasma nitriding in the same equipment for AISI 4140 steel, and plain air was used as the oxygen bearing gas. The cross-sectional microstructures of the treated samples were observed by optical metallography and scanning electron microcopy (SEM), and the thickness of compound layer was measured accordingly. The phases were determined by X-ray diffraction (XRD), corrosion resistance was evaluated by electrochemical polarization, and the surface morphology before and after polarization test was also observed by SEM. Meanwhile, standard Gibbs free energy of the oxidation reactions existed in Fe–O system was calculated. The results show that a thin iron oxide layer composed of magnetite (Fe 3 O 4 ) and hematite (Fe 2 O 3 ) is formed on top of the compound layer during plasma post-oxidizing process, and the ratio of magnetite (Fe 3 O 4 ) to hematite (Fe 2 O 3 ) is depended on plasma post-oxidizing temperature and time. Highest ratio of Fe 3 O 4 to Fe 2 O 3 is obtained while post-oxidizing at 673 K for 60 min due to lower standard Gibbs free energy and appropriate forming rate for the formation of Fe 3 O 4 at this temperature. The thin oxide layer brings out significant enhancement of corrosion resistance, especially at higher ratios of Fe 3 O 4 to Fe 2 O 3 , due to the dense and adherent characteristic of Fe 3 O 4 oxide. Surface images of the post-oxidizing specimen show that a dense oxide layer was formed when post-oxidized at 673 K for 60 min, and those after polarization test show that there exists no corrosion pit for the specimen post-oxidized at 673 K for 60 min, however some corrosion pits occur for those treated under other conditions. Meanwhile, the thickness of the compound layer formed during nitriding process tends to decrease with the increase of post-oxidizing temperature and time due to spallation by air plasma bombardment during post-oxidizing process.

AISI 4140 steel was selected as the material and plasma post-oxidation by plain air in the same nitriding system just after plasma nitriding was investigated primarily. The results show that a thin oxide layer composed of Fe 3 O 4 and Fe 2 O 3 is formed, and the ratio of Fe 3 O 4 to Fe 2 O 3 is closely related to the plasma post-oxidizing temperature and time. The highest ratio of Fe 3 O 4 to Fe 2 O 3 is obtained while post-oxidizing at 673 K for 60 min due to lower standard Gibbs free energy and appropriate forming rate for the formation of Fe 3 O 4 at this condition. The thin oxide layer brings out significant improvement of corrosion resistance, especially at higher ratio of Fe 3 O 4 to Fe 2 O 3 . Surface images of the corroded specimens after polarization test shows that there exists no corrosion pit for the specimen post-oxidized at 673 K for 60 min, though some corrosion pits exist for those treated under other conditions.

Propane was used as carbon-bearing gas in plasma nitrocarburizing due to its lower ionization energy and the effect of its concentration on the nitrocarburizing rate and phase compositions for AISI 1045 steel was investigated. The surfaces of nitrocarburized samples were analysed by optical microscopy, X-ray diffraction and by using a microhardness tester. The results showed that the case depth increased initially with the increasing concentration of propane, and then decreased when it exceeded a limit of 1.5%. The maximum case depth of 40 μm was achieved at this optimum concentration of 1.5% when nitrocarburizing at 510 °C for 4 h, and a maximum surface hardness of 7.63 GPa and desirable phases of dominant e nitride with a little cementite were obtained. The amount of cementite increased with the increasing concentration of propane above 1.5%, which hindered the nitrogen and carbon up-take, thus resulting in the decrease of case depth and surface hardness.

The multi-target kinase inhibitor sorafenib has been approved for the treatment of patients with advanced differentiated thyroid cancer. However, different sensitivities to sorafenib have been observed, and few patients have benefited from sorafenib treatment in the long term. In the event of acquired resistance to sorafenib it is not beneficial to continue treatment in most patients. Autophagy can be induced in a variety of cancer treatments and plays an important role in cancer treatment. The role of autophagy in sorafenib treatment of thyroid cancer has not been fully demonstrated. The present study investigated whether autophagy is activated by sorafenib during the treatment of thyroid cancer, examined the underlying mechanisms, and explored potential strategies to enhance the therapeutic sensitivity of sorafenib. Chloroquine (CQ) is an autophagy inhibitor that has been reported to increase sensitivity to various cancer treatments. Thyroid cancer xenograft model mice were treated with sorafenib, CQ, or a combination of sorafenib and CQ. We observed that CQ or sorafenib treatment suppressed tumor growth, while mice treated with the combination of sorafenib and CQ displayed significantly reduced tumor growth compared with those treated with sorafenib or CQ alone. Western blotting results indicated that sorafenib concurrently inhibited the activities of the MAPK and AKT/mTOR pathways in thyroid cancer. Autophagy was activated by sorafenib in thyroid cancer, both in vitro and in vivo, which was at least in part due to suppression of the AKT/mTOR pathway. Combination treatment including CQ could inhibit the autophagic flux induced by sorafenib. Silencing the key autophagy gene ATG5 using small interfering RNA also increased the anticancer effect of sorafenib. In summary, the present study revealed that inhibition of autophagy enhances the anticancer effect of sorafenib, and the combination of CQ with sorafenib treatment represents a potential therapeutic strategy for treating advanced differentiated thyroid cancer.

Sorafenib is a multikinase inhibitor that has been approved for the treatment of patients with advancedThe thyroid cancer cell lines 8505C and FTC133 were treated with sorafenib in the presence or absence of BEZ235 or small interfering RNA (siRNA) directed against AKT. A CCK8 kit was used to evaluate cell viability. Protein expression levels of relevant genes were determined by Western blotting analysis, whereas messenger RNA expression levels were determined by real-time PCR analysis. Flow cytometry was performed to assess the number of apoptotic cells.The results indicate that sorafenib simultaneously inhibited the activities of the MAPK and PI3K/AKT/mTOR pathways in thyroid cancer cells. Treatment of 8505C and FTC133 cells with NVP-BEZ235, siRNA against AKT, or sorafenib induced tumor cell apoptosis and led to reduced tumor cell proliferation. Sorafenib in combination with PI3K/AKT/mTOR inhibition by NVP-BEZ235 or AKT siRNA enhanced apoptosis and proliferation suppression.The evidence of this study suggests that a combinatorial approach that inhibits both the MAPK and PI3K/AKT/mTOR pathways exerts a greater antitumor effect than sorafenib alone in thyroid cancer cell lines.

The sharply increasing incidence of thyroid cancer has attracted considerable attention over the last few years. The combination of surgery, radioiodine ablation and thyroid-stimulating hormone suppression is usually efficient for the majority of thyroid tumors. However, advanced thyroid cancer that is recurrent, metastatic and 131I-refractory, or medullary thyroid cancer, pose a therapeutic challenge. Autophagy is a process that metabolizes damaged cytoplasmic organelles and long-lived proteins in order to recycle cellular materials and maintain homeostasis. It has been confirmed that autophagy plays a dual role during cancer development, progression and treatment, mainly depending on the type and stage of the tumor. Autophagy modulation has become a potential therapeutic target for diverse diseases. The mechanism of thyroid tumorigenesis and cancer progression was largely demonstrated to be correlated with the dysregulation of the Ras/Raf/mitogen-activated protein kinase kinase/extracellular signal-regulated kinase and the phosphoinositide 3-kinase/Akt/mammalian target of rapamycin pathways, as well as with abnormal epigenetic modifications. Those mechanisms are associated with autophagy regulation and may be beneficial for the treatment of advanced thyroid cancer. However, the number of available studies on the role of autophagy in thyroid cancer development, progression and treatment outcome, is currently limited. The aim of this review was to elaborate on the relevant knowledge and future prospectives of autophagy in the treatment of thyroid cancer.

Background Omega-3 polyunsaturated fatty acids (PUFAs), including alpha-linolenic acid (ALA), eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA), and docosapentaenoic acid (DPA), play critical roles in numerous biochemical reactions. Our aim is to develop a rapid and sensitive method for simultaneous determination of ALA, EPA, DHA and DPA in the plasma of hyperlipidemic and normolipidemic subjects. Methods An ultra-high-performance liquid chromatography-tandem mass spectrometric (UPLC-MS/MS) method of ALA, EPA, DHA, and DPA was developed with chlorzoxazone as the internal standard (IS). The analytes were separated on an Acquity BEH C18 column (2.1 mm × 100 mm, 1.7 μm) with gradient elution by acetonitrile and 0.1% ammonia water. ALA, EPA, DHA, DPA, and IS were determined by negative electrospray ionization (ESI-) with multiple reaction monitoring (MRM) at m/z 277.42/259.05, 301.20/257.00, 327.30/283.40, 329.24/285.32, and 168.03/132.02. A total of 80 normolipidemic subjects and 83 hyperlipidemic subjects, who underwent testing for plasma lipids, liver and kidney functions, and blood routine blood test (BRT), were enrolled. Results There was good linearity for ALA within 1–10 μg/mL, and EPA, DHA and DPA were within 0.125–10 μg/mL. The relative standard deviation (RSD) of precision was below 15%. The concentrations of ALA, EPA, DHA and DPA were 3.47 ± 2.58, 0.41 ± 0.26, 2.93 ± 1.39 and 0.25 ± 0.21 μg/mL, respectively, in normolipidemic subjects, increasing to 4.14 ± 3.71, 0.57 ± 0.46, 3.43 ± 2.13, 0.27 ± 0.25 μg/mL, respectively in hyperlipidemic subjects. Among them, only the EPA concentration was significantly different between two groups. There was a high correlation between ALA, EPA, DHA and DPA. Conclusion We developed a rapid and sensitive method for simultaneously determination of ALA, EPA, DHA and DPA in hyperlipidemic and normolipidemic subjects. In hyperlipidemic and normolipidemic subjects, concentrations of ALA were highest, followed by DHA, EPA and DPA; there were high degrees of correlation between each value.

In teleoperation, the operator is often required to command the motion of the remote robot and monitor its behavior. However, such an interaction demands a heavy workload from a human operator when facing with complex tasks and dynamic environments. In this article, we propose a shared control method to assist the operator in the manipulation tasks to reduce the workload and improve the efficiency. We adopt a task-parameterized hidden semi-Markov model to learn a manipulation skill from several human demonstrations. We utilize the learned model to predict the manipulation target given the current observed robotic motion trajectory and subsequently estimate the desired robotic motion given the current input of the operator. The estimated robotic motion is then utilized to correct the input of the operator to provide manipulation assistance. In addition, a set of virtual reality devices are used to capture the operator’s motion and display the vision feedback from the remote site. We evaluate our approach through two manipulation tasks with a dual-arm robot. The experimental results show the effectiveness of the proposed method.

In some nonlinear dynamic systems, the state variables function usually can be separated from the control variables function, which brings much trouble to the identification of such systems. To well solve this problem, an improved least squares support vector regression (LSSVR) model with multiple-kernel is proposed and the model is applied to the nonlinear separable system identification. This method utilizes the excellent nonlinear mapping ability of Morlet wavelet kernel function and combines the state and control variables information into a kernel matrix. Using the composite wavelet kernel, the LSSVR includes two nonlinear functions, whose variables are the state variables and the control ones respectively, in this way, the regression function can gain better nonlinear mapping ability, and it can simulate almost any curve in quadratic continuous integral space. Then, they are used to identify the two functions in the separable nonlinear dynamic system. Simulation results show that the multiple-kernel LSSVR method can greatly improve the identification accuracy than the single kernel method, and the Morlet wavelet kernel is more efficient than the other kernels.

The purpose of this study is to examine the effectiveness of introducing both rituximab (RTX) and 131I for active Graves' ophthalmopathy (GO) with hyperthyroidism.In total, 217 patients suffering from active GO with hyperthyroidism were included in this research. All subjects were randomly assigned to 3 groups. Patients in group A solely received 131I treatment; group B1 underwent a methylprednisolone treatment in combination with 131I treatment; and group B2 received an RTX in combination with 131I treatment. Hyperthyroidism treatment outcomes, orbital volumetry, ophthalmic assessments, serum cytokine levels, and adverse effects were measured after treatment.The orbital volumetry principle was significantly different from 24 weeks after the start of treatment among all 3 groups, and improvements in most ophthalmic parameters were regarded significantly different among 3 groups (all p0.05). The expression levels of miR-146a and most serum cytokines were regarded significantly different from 24 weeks after the start of treatment among 3 groups (all p0.05).In comparison with other therapies, RTX treatment in combination with 131I treatment is considered to be more effective for hyperthyroidism with active GO.

Lung carcinoma is the leading cause of cancer-related deaths and is the primary source for choroidal metastasis in over 20% cases. Non-small-cell lung cancer (NSCLC) accounts for 85% of all lung cancer cases. Patients with metastatic NSCLC have a median survival of one year. Successful treatment of systemic metastasis from NSCLC using erlotinib has been documented. The effect of oral erlotinib on choroidal metastasis has been rarely reported. We document a case and study the effect of oral erlotinib on choroidal metastasis from NSCLC. A 48-year-old Caucasian female presented with biopsy-proven primary NSCLC with systemic metastasis and solitary choroidal metastasis of 4.8 mm thickness in the right eye. The patient was treated with 100 mg daily dose of oral erlotinib. Two weeks after starting erlotinib therapy, the patient showed complete regression of choroidal metastasis to a flat scar with resolution of subretinal fluid and improvement of visual acuity from 20/100 to 20/25. There was no evidence of recurrence at five-month follow-up. Erlotinib is an alternative therapy for choroidal metastasis from NSCLC.

Differentiated thyroid carcinoma (DTC) is the most common endocrine malignancy. Surgical removal with radioactive iodine therapy is recommended for recurrent thyroid carcinoma, and the postsurgical thyroid removal is critical. This study evaluated the clinical values of radiofrequency ablation (RFA) in the postsurgical thyroid removal for DTC. 35 DTC patients who had been treated by subtotal thyroidectomy received RFA for postsurgical thyroid removal. Before and two weeks after RFA, the thyroid was examined by ultrasonography and 99mTcO4 - thyroid imaging, and the serum levels of free triiodothyronine (FT3), free thyroxin (FT4), thyroid stimulating hormone (TSH) and thyroglobulin (Tg) were detected. The efficacy and complications of RFA were evaluated. Results showed that, the postsurgical thyroid removal by RFA was successfully performed in 35 patients, with no significant complication. After RFA, the average largest diameter and volume were significantly decreased in 35 patients (P > 0.05), and no obvious contrast media was observed in ablation area in the majority of patients. After RFA, the serum FT3, FT4 and Tg levels were markedly decreased (P < 0.05), and TSH level was significantly increased (P < 0.05). After RFA, radioiodine concentration in the ablation area was significantly reduced in the majority of patients. The reduction rate of thyroid update was 0.69±0.20%. DTC staging and interval between surgery and RFA had negative correlation (Pearson coefficient = -0.543; P = 0.001), with no obvious correlation among others influential factors. RFA is an effective and safe method for postsurgical thyroid removal of DTC.

Background. Induction of hyperthermia by radiofrequency ablation is gaining popularity in treating a variety of solid tumors. This study examined an impact of sublethal heat treatment interacted with chemotherapeutic drugs on the survival of head and neck squamous carcinoma cells using in vitro model. Materials and methods. FaDu cells were subjected to heat treatment at 42°C or 45°C for 15 min either before or after exposure to cisplatin or hydroxyurea. The survival of cells was determined by 3-(4,5-dimethylthiazolyl-2)-2,5-diphenyltetrazolium bromide assay. The RNA and protein levels of various heat shock proteins were examined by reverse transcription polymerase chain reaction and Western blot analysis, respectively. Cell cycle progression was analyzed by flow cytometry with propidium iodide staining. Results. FaDu cells preheated to 45°C exhibited an increased resistance to hydroxyurea but not to cisplatin. The heat treatment resulted in induction of HSP70 expression at transcript and protein levels, but there was no change in expression of HSP90β and HSP27. After heat treatment, cells accumulated in S-phase at 3 h and proceeded to G 2 /M phase at 24 h. When cells pre-exposed to drugs for 24 h, the cisplatin-treated cells exhibited a higher thermotolerance than the hydroxyurea-treated cells at heat treatment of 45°C. Cisplatin and hydroxyurea caused cells to accumulate in S-phase and increased the protein expression of HSP27 but not HSP90β and HSP70. Conclusion. FaDu cells surviving the heat treatment expressed HSP70 and disrupted cell cycle progression, which resulted in developing a resistance to subsequent hydroxyurea treatment. However, the heat treatment did not have an effect on the sensitivity to cisplatin. In the reversed procedure, pre-exposure to hydroxyurea and cisplatin resulted in developing a thermotolerance.

As an important parameter for critical load calculation and soil acidification simulation, weathering rates of soils in China were studied using different methods of calculation. The approaches used were the mass balance approach, the soil mineralogical classification, the total analysis correlation, the PROFILE model, the MAGIC model and a simulated leaching experiment. Since chemical weathering of secondary minerals usually plays a much more important role in neutralizing the long-term acidification of soils in China than that of parent material, soil mineralogy rather than parent rock/material type, which is regarded as the most suitable factor representing weathering rates in Europe, should be adopted as the basis for soil classification. The weathering rate assigned to each soil should also be corrected when the effect of temperature is considered. Due to the variation in experimental conditions, the weathering rates of soils from laboratory experiment may be difficult to compare with field determined rates, and should be adjusted by pH and percolation rate. The comparison of various methods in this study shows that the weathering rates of soils estimated by the PROFILE model coincide well with those from other independent methods such as the dynamic modeling by MAGIC and the modified leaching experiment. The weathering rates were very low (usually lower than 1.0 kEq·ha−1·year−1) for Allites (including Latosol, Lateritic Red Earth, Red Earth, Yellow Earth and Yellow-Brown Earth) in south China and Silalsols (consisting of Dark Brown Forest Soil, Black Soil and Podzolic Soil) in northeast China, and very high for Alpine Soils, Desert Soils and Pedocals in west China. The content of weatherable minerals in soil is the most important factor in determining the spatial distribution of weathering rate in China, while the difference in temperature may be the reason why the weathering rate of soil in northeast China was lower than that in southeast China.