Decreased TAX1BP1 participates in systemic lupus erythematosus by regulating monocyte/macrophage function.

Systemic lupus erythematosus (SLE) involves disorders of innate and adaptive immune pathways. Tax1-binding protein 1 (TAX1BP1) modulates the production of antibodies in B cells and the T-cell cycle by regulating the NF-κB signaling pathway. However, the potential association of TAX1BP1 with SLE and its role in monocytes/macrophages have not been fully elucidated. In this study, we utilized whole-exome sequencing (WES) in combination with Sanger sequencing and identified 16 gene mutations, including in TAX1BP1, in an SLE family. TAX1BP1 protein expression with western blotting detection was reduced in SLE patients and correlated with disease activity negatively. Furthermore, RNA sequencing and 4D Label-Free Phosphoproteomic analysis were employed to characterize the transcriptome and phosphoproteome profiles in THP-1 and THP-1-differentiated M1 macrophages with TAX1BP1 knockdown. Silencing of TAX1BP1 in THP-1 and THP-1-differentiated M1 macrophages led to an increase in cluster of differentiation 80 (CD80) expression and differential changes in CD14 and CD16 expression, as assessed by flow cytometry. Additionally, western blot analysis showed that knockdown of TAX1BP1 led to a reduction in TRAF6 and p-p65 in THP-1-differentiated macrophages, with or without lipopolysaccharide (LPS) or tumor necrosis factor (TNF)-α stimulation. Taken together, our findings suggest that TAX1BP1 participates in SLE activity by regulating antigen presentation in monocytes and inflammatory responses in M1 macrophages.

Transcriptome Analysis Reveals the Mechanism of Exogenous Selenium in Alleviating Cadmium Stress in Purple Flowering Stalks (Brassica campestris var. purpuraria).

In China, cadmium (Cd) stress has a significant role in limiting the development and productivity of purple flowering stalks (Brassica campestris var. purpuraria). Exogenous selenium supplementation has been demonstrated in earlier research to mitigate the effects of Cd stress in a range of plant species; nevertheless, the physiological and molecular processes by which exogenous selenium increases vegetable shoots' resistance to Cd stress remain unclear. Purple flowering stalks (Brassica campestris var. purpuraria) were chosen as the study subject to examine the effects of treatment with sodium selenite (Na2SeO3) on the physiology and transcriptome alterations of cadmium stress. Purple flowering stalk leaves treated with exogenous selenium had higher glutathione content, photosynthetic capacity, and antioxidant enzyme activities compared to the leaves treated with Cd stress alone. Conversely, the contents of proline, soluble proteins, soluble sugars, malondialdehyde, and intercellular CO2 concentration tended to decrease. Transcriptome analysis revealed that 2643 differentially expressed genes (DEGs) were implicated in the response of exogenous selenium treatment to Cd stress. The metabolic pathways associated with flavonoid production, carotenoid synthesis, glutathione metabolism, and glucosinolate biosynthesis were among those enriched in these differentially expressed genes. Furthermore, we discovered DEGs connected to the production route of glucosinolates. This work sheds fresh light on how purple flowering stalks' tolerance to cadmium stress is improved by exogenous selenium.

Integrated network pharmacology to investigate the mechanism of Salvia miltiorrhiza Bunge in the treatment of myocardial infarction.

Salvia miltiorrhiza Bunge is a natural drug for treating myocardial infarction (MI). However, the targets and mechanisms of S. miltiorrhiza Bunge in the treatment of MI are yet to be elucidated. Traditional Chinese medicine systems pharmacology (TCMSP) data were used to screen out chemical constituents, and UniProt was used to predict relevant targets. Disease targets were obtained from the Online Mendelian Inheritance in Man and GeneCards databases. We used the STRING platform to build a protein-protein interaction network and used Cytoscape_v3.8.1 software to make a Drug-Ingredients-Gene Symbols-Disease network map. The Metascape database was used to perform gene ontology and Kyoto Encyclopaedia of Genes and Genomes (KEGG) enrichment analyses for drug-disease overlapping gene symbols. The targets identified by network pharmacology were further verified by in vitro and in vivo experiments. Seventy-five active components of S. miltiorrhiza Bunge were obtained from the TCMSP database, while 370 disease targets and 29 cross-targets were obtained from the Genecards database. The KEGG pathway enrichment results suggested that the mechanism of S. miltiorrhiza Bunge in the treatment of MI was significantly related to the VEGF signalling pathway. In vitro and in vivo experiments were used to evaluate the reliability of some important active ingredients and targets. S. miltiorrhiza Bunge alleviated the damage to cardiac function, attenuated myocardial fibrosis and protected endothelial cell function by increasing the expression of TGF-ß and VEGFA. S. miltiorrhiza Bunge showed the therapeutic effect of MI by promoting the expression of VEGFA signalling pathway, providing a reliable basis for exploring herbal treatment of MI.

Genetic diversity and gene expression diversity shape the adaptive pattern of the aquatic plant Batrachium bungei along an altitudinal gradient on the Qinghai-Tibet plateau.

It is an intriguing issue of evolutionary biology how genetic diversity and gene expression diversity shape the adaptive patterns. Comparative transcriptomic studies of wild populations in extreme environments provide critical insights into the relative contribution of genetic and expressive components. In this study, we analyzed the genetic diversity and gene expression diversity of 20 populations of the aquatic plant Batrachium bungei along elevations ranging from 2690 to 4896 m on the Qinghai-Tibet plateau (QTP). Based on single nucleotide polymorphisms (SNPs) and gene expression data from 100 individuals of B. bungei, we found that variation in genetic sequence was more sensitive to detect weak differentiation than gene expression. Using 292,613 high-quality SNPs, we documented a significant phylogeographical structure, a low within-population genetic diversity, and a high inter-population genetic differentiation in B. bungei populations. Analysis of relationship between geographic distance, genetic distance, and gene expression similarity showed that geographic isolation shaped gene flow patterns but not gene expression patterns. We observed a negative relationship between genetic diversity and gene expression diversity within and among B. bungei populations, and we demonstrated that as environmental conditions worsen with increasing altitude, genetic diversity played an increased role in maintaining the stability of populations, while the corresponding role of gene expression diversity decreased. These results suggested that genetic diversity and gene expression diversity might act as a complementary mechanism contributing to the long-term survival of B. bungei in extreme environments.

Prevalence and genetic variation of the M, N, and S2 genes of feline coronavirus in Shandong Province, China.

Feline coronavirus (FCoV) is the causative agent of feline infectious peritonitis and diarrhoea in kittens worldwide. In this study, a total of 73 feline diarrhoeal faecal samples were collected from animal hospitals and pet markets in ShanDong province from 2017 to 2019. FCoV was detected in 58.23% (46/73) of the samples, using the RT-PCR method. The results showed that the detection rate of FCoV in healthy cats and sick cats was 41.7% (10/24) and 81.6% (40/49), respectively. Full gene amplification and sequencing of the N, M, and S2 genes of FCoV isolates were performed. An amino acid mutation (M1058L) in the S2 gene was found that can be used as a marker for distinguishing feline enteric coronavirus (FECV) from feline infectious peritonitis virus (FIPV). This study provides new epidemiological information about FCoV that will aid in the prevention of FCoV in China.

Multisequence magnetic resonance imaging-based radiomics models for the prediction of microsatellite instability in endometrial cancer.

PURPOSE: To evaluate the performance of multisequence magnetic resonance imaging (MRI)-based radiomics models in the assessment of microsatellite instability (MSI) status in endometrial cancer (EC). MATERIALS AND METHODS: This retrospective multicentre study included 338 EC patients with available MSI status and preoperative MRI scans, divided into training (37 MSI, 123 microsatellite stability [MSS]), internal validation (15 MSI, 52 MSS), and external validation cohorts (30 MSI, 81 MSS). Radiomics features were extracted from T2-weighted images, diffusion-weighted images, and contrast-enhanced T1-weighted images. The ComBat harmonisation method was applied to remove intrascanner variability. The Boruta wrapper algorithm was used for key feature selection. Three classification algorithms, logistic regression (LR), random forest (RF), and support vector machine (SVM), were applied to build the radiomics models. The area under the receiver operating characteristic curve (AUC) was calculated to compare the diagnostic performance of the models. Decision curve analysis (DCA) was conducted to determine the clinical usefulness of the models. RESULTS: Among the 1980 features, Boruta finally selected nine radiomics features. A higher MSI prediction performance was achieved after running the ComBat harmonisation method. The SVM algorithm had the best performance, with AUCs of 0.921, 0.903, and 0.937 in the training, internal validation, and external validation cohorts, respectively. The DCA results showed that the SVM algorithm achieved higher net benefits than the other classifiers over a threshold range of 0.581-0.783. CONCLUSION: The multisequence MRI-based radiomics models showed promise in preoperatively predicting the MSI status in EC in this multicentre setting.

Radiomics based on multisequence magnetic resonance imaging for the preoperative prediction of peritoneal metastasis in ovarian cancer.

OBJECTIVES: To develop a radiomics signature based on multisequence magnetic resonance imaging (MRI) to preoperatively predict peritoneal metastasis (PM) in ovarian cancer (OC). METHODS: Eighty-nine patients with OC were divided into a training cohort including patients (n = 54) with a single lesion and a validation cohort including patients (n = 35) with bilateral lesions. Radiomics features were extracted from the T2-weighted images (T2WIs), fat-suppressed T2WIs, multi-b-value diffusion-weighted images (DWIs), and corresponding parametric maps. A radiomics signature and nomogram incorporating the radiomics signature and clinical predictors were developed and validated on the training and validation cohorts, respectively. RESULTS: The radiomics signature generated by 6 selected features showed a favorable discriminatory ability to predict PM in OC with an area under the curve (AUC) of 0.963 in the training cohort and an AUC of 0.928 in the validation cohort. The nomogram, comprising the radiomics signature, pelvic fluid, and CA-125 level, showed more favorable discrimination with an AUC of 0.969 in the training cohort and 0.944 in the validation cohort. Net reclassification index with values of 0.548 in the training cohort and 0.500 in the validation cohort. CONCLUSION: Radiomics signature based on multisequence MRI serves as an effective quantitative approach to predict PM in OC patients. A nomogram of radiomics signature and clinical predictors could further improve the prediction ability of PM in patients with OC. KEY POINTS: • Multisequence MRI-based radiomics showed a favorable discriminatory ability to predict PM in OC. • The nomogram incorporating the radiomics signature and clinical predictors was clinically useful to preoperatively predict PM in patients with OC.

Radiomics derived from dynamic contrast-enhanced MRI pharmacokinetic protocol features: the value of precision diagnosis ovarian neoplasms.

OBJECTIVES: To evaluate the efficiency of 2- and 3-class classification predictive tasks constructed from radiomics features extracted from dynamic contrast-enhanced magnetic resonance imaging (DCE-MRI) pharmacokinetic (PK) protocol in discriminating among benign, borderline, and malignant ovarian tumors. METHODS: One hundred and four ovarian lesions were evaluated using preoperative DCE-MRI. Radiomics features were extracted from 7 types of DCE-MR images. To explore the differential ability of radiomics between three types of ovarian tumors, two- and three-class classification tasks were established. The 2-class classification task was divided into three subtasks: benign vs. borderline (task A), benign vs. malignant (task B), and borderline vs. malignant (task C). For the 3-class classification task, 104 lesions were randomly divided into training (72 lesions) and validation (32 lesions) cohorts. The discrimination abilities of the radiomics signatures were established with the training cohort and tested with the independent validation cohort. The predictive performance of the task was evaluated by receiver operating characteristic (ROC) curve, calibration curve analysis, and decision curve analysis (DCA). RESULTS: For the 2-class classification task, the combination of PK radiomics signatures model (PK model) showed a good diagnostic ability with the highest area under the ROC curves (AUCs) of 0.899, 0.865, and 0.893 for tasks A, B, and C, respectively. Additionally, the 3-class classification task demonstrated a good discrimination performance with AUCs of 0.893, 0.944, and 0.891 for the benign, borderline, and malignant groups, respectively. CONCLUSIONS: Radiomics analysis based on the DCE-MRI PK protocol showed promise for discriminating among benign, borderline, and malignant ovarian tumors. KEY POINTS: • Two-class classification predictive task of DCE-MRI PK protocol enabled the classification of 3 categories of ovarian tumors through the pairwise comparison strategy with a perfect diagnostic ability. • Three-class classification predictive task maintained good performance to effectively judge each category of ovarian tumors directly.

Regulatory roles of differentially expressed MicroRNAs in metabolic processes in negative Lens-induced myopia Guinea pigs.

BACKGROUND: Myopia is one of the most common vision defects worldwide. microRNAs can regulate the target gene expression, influencing the development of diseases. RESULTS: To investigate the alterations of microRNA profiling in negative lens-induced myopia (NLIM) guinea pigs and to explore the regulatory role of microRNAs in the occurrence and the development of myopia, we first established the NLIM guinea pig model after induction for 2 weeks. Further, we isolated sclera to purify total messenger RNA (mRNA) in both NLIM and NLIM fellow sclera. Using next generation sequencing technique and bioinformatics analysis, we identified the differentially expressed microRNAs in NLIM guinea pigs, performed the bioinformatics annotation for the differentially expressed microRNAs, and validated the expression of differentially expressed microRNAs. As a result, we successfully established an NLIM model in guinea pigs, identified 27 differentially expressed microRNAs in NLIM guinea pig sclera, including 10 upregulated and 17 downregulated microRNAs. The KEGG annotation showed the main signaling pathways were closely associated with PPAR signaling, pyruvate and propanoate metabolisms, and TGF-beta signaling pathways. CONCLUSIONS: Our findings indicate that the development of myopia is mainly involved in the disorder of metabolic processes in NLIM guinea pigs. The PPAR signaling, pyruvate and propanoate metabolism pathways may play roles in the development of myopia.

Intravoxel Incoherent Motion Diffusion-weighted MRI of Infiltrated Marrow for Predicting Overall Survival in Newly Diagnosed Acute Myeloid Leukemia.

Background Acute myeloid leukemia (AML) features relatively low overall survival (OS). Intravoxel incoherent motion (IVIM) diffusion-weighted MRI separates tissue microcapillary perfusion and diffusivity and may have potential for helping to assess prognosis in infiltrated marrow disease apart from solid tumor. Thus, a study of overall survival would contribute to clarifying the value of IVIM for assessing long-term prognosis in AML. Purpose To determine whether the IVIM-derived parameters of infiltrated bone marrow may be associated with OS in newly diagnosed AML. Materials and Methods This prospective study enrolled participants with newly diagnosed AML between July 2014 to March 2016 consecutively. Participants underwent MRI of the lumbar spine by using an IVIM sequence. Participant clinical characteristics and OS were collected. The median of follow-up period was 20 months (range, 1-56 months). The IVIM parameters (pseudoperfusion fraction, f; diffusion coefficient, D; and pseudodiffusion coefficient, D*) were obtained. A nonparametric log-rank test was used to identify the threshold of IVIM parameters for OS. Univariable Kaplan-Meier and multivariable Cox proportional hazards regression analyses were performed to investigate prognostic significance of possible indicators. Results Fifty-three participants (mean age, 42 years ± 17; 30 men) were evaluated. Nonparametric log-rank test results showed that the thresholds of f and D values for OS were 31.0% and 0.2 × 10-3 mm2/sec, respectively. Univariable analyses indicated that high f value (>31.0%) and low D value (≤0.2 × 10-3 mm2/sec) were associated with shorter OS (P = .003 and .01, respectively). An f value greater than 31.0% (hazard ratio, 2.4; 95% confidence interval: 1.0, 5.6; P = .046) was associated with OS, independent of clinical confounders (age, karyotype, and white blood cell counts) in a multivariable analysis. Conclusion Pseudoperfusion fraction and diffusion coefficient from intravoxel incoherent motion diffusion-weighted MRI may be viable prognosis predictors of newly diagnosed acute myeloid leukemia. © RSNA, 2020.

Intravoxel Incoherent Motion Imaging in Differentiation Borderline From Malignant Ovarian Epithelial Tumors: Correlation With Histological Cell Proliferation and Vessel Characteristics.

BACKGROUND: The differentiation of borderline from malignant ovarian epithelial tumors (OETs) is difficult based on morphologic characteristics. Diffusion and perfusion information from intravoxel incoherent motion (IVIM) diffusion-weighted imaging (DWI) might be useful for this distinction. PURPOSE: To investigate the potential of IVIM-DWI in discriminating borderline from malignant OETs by correlating with cell proliferation and microvessel density (MVD). STUDY TYPE: Prospective. SUBJECTS: Sixty-six patients with OETs (22 borderline, BOETs; 44 malignant, MOETs) underwent IVIM-DWI before surgery. FIELD STRENGTH: 3.0T IVIM-DWI sequence with 12 b-values (0-1000 sec/mm2 ). ASSESSMENT: Apparent diffusion coefficient (ADC) and IVIM-DWI parameters (diffusion coefficient [D], microvascular volume fraction [f], and pseudodiffusion coefficient [D*]) were measured. Cell proliferation and MVD was determined by immunohistochemical staining of Ki-67 and CD-31, respectively. STATISTICAL TESTS: Mann-Whitney U-test; two-sample t-test; intraclass correlation coefficient; Bland-Altman analysis; receiver operating characteristics (ROC) curves; and Spearman correlation. RESULTS: ADC and D in BOETs was significantly higher than those in MOETs (P < 0.001), while f in BOETs was significantly lower than those in MOETs (P < 0.001). No significant difference was found in D* between borderline and malignancies (P = 0.324). In the differential diagnosis of borderline from malignant OETs; D demonstrated the highest area under the curve (AUC) of 0.951, while ADC and f had a lower AUC of 0.921 and 0.847, respectively. The ADC and D was negatively correlated with cell proliferation (r = -0.682, r = -0.694, respectively, P < 0.001), while f was positively correlated with MVD of the OETs (r = 0.558, P < 0.001). DATA CONCLUSION: IVIM-DWI may be a useful tool for differentiating BOETs from MOETs. D and f could be image biomarkers to reflect histopathological characteristics of cell proliferation and MVD in OETs. LEVEL OF EVIDENCE: 1 Technical Efficacy Stage: 2 J. Magn. Reson. Imaging 2020;51:928-935.

Correlation of Intravoxel Incoherent Motion Parameters and Histological Characteristics From Infiltrated Marrow in Patients With Acute Leukemia.

BACKGROUND: An accurate and noninvasive method is of great importance to assess angiogenesis and cellularity of bone marrow in acute leukemia (AL). PURPOSE: To investigate whether the intravoxel incoherent motion (IVIM) parameters correlate with the histological characteristics of infiltrated marrow in AL patients and compare the difference between acute lymphoblastic leukemia (ALL) and acute myeloid leukemia (AML). STUDY TYPE: Prospective. POPULATION MODEL: Forty newly diagnosed patients with AL, including 20 AML and 20 ALL. FIELD STRENGTH/SEQUENCE: 1.5T/T1 WI and IVIM. ASSESSMENT: IVIM-derived parameters (true diffusion coefficient D, pseudodiffusion coefficient D*, and perfusion fraction, f) were measured in lumbar marrow. Histopathological analyses were performed from samples of marrow biopsy. STATISTICAL TESTS: The correlations between IVIM parameters and histological parameters used the Spearman correlation test. The difference of IVIM parameters and histological parameters between ALL and AML groups used the unpaired t-test or Mann-Whitney U-test, as appropriate. RESULTS: The f was positively correlated with microvessel density (MVD) in patients with ALL, AML, and AL (r = 0.740, P = 0.006; r = 0.771, P < 0.001; and r = 0.750, P < 0.001, respectively). There were no significant correlations between D and bone marrow cellularity in the three groups (r = -0.289, P = 0.362; r = 0.281, P = 0.292; and r = 0.058, P = 0.769, respectively). D and f values of ALL were higher than that of AML group (t = 3.332, P = 0.003 and t = 2.600, P = 0.014, respectively). MVD was higher in ALL than AML (t = 2.120, P = 0.044), whereas bone marrow cellularity had no significant difference between AML and ALL (t = -0.682, P = 0.501). DATA CONCLUSION: The f value derived from IVIM in bone marrow was positively correlated with MVD, while f might be able to show a difference of vascularity between ALL and AML. Therefore, the f value can be used as an alternative imaging marker of angiogenesis in marrow of AL patients. LEVEL OF EVIDENCE: 1 Technical Efficacy Stage: 3 J. Magn. Reson. Imaging 2020;51:1720-1726.

RETRACTED: Notoginsenoside R1 suppresses miR-301a via NF-κB pathway in lipopolysaccharide-treated ATDC5 cells.

This article has been retracted: please see Elsevier Policy on Article Withdrawal (https://www.elsevier.com/about/our-business/policies/article-withdrawal). This article has been retracted at the request of the Editor-in-Chief. The journal was initially contacted by the corresponding author to request the retraction of the article as the data were not reliable. Given the comments of Dr Elisabeth Bik regarding this article " … the Western blot bands in all 400+ papers are all very regularly spaced and have a smooth appearance in the shape of a dumbbell or tadpole, without any of the usual smudges or stains. All bands are placed on similar looking backgrounds, suggesting they were copy/pasted from other sources, or computer generated", the journal requested the authors to provide the raw data. However, the authors were not able to fulfil this request and therefore the Editor-in-Chief decided to retract the article.

Effects of verapamil on the pharmacokinetics of puerarin in rats.

The oral bioavailability of puerarin is poor which hindered its clinical performance. This study investigates the effects of verapamil on the pharmacokinetics of puerarin in rats. The pharmacokinetics of orally administered puerarin (50 mg/kg) with or without verapamil pretreatment (10 mg/kg/day for 7 days) were investigated. The plasma concentration of puerarin was determined using LC-MS/MS method, and the pharmacokinetics profiles were calculated and compared. Caco-2 cell transwell model was also used to investigate the effects of verapamil on the transport pf puerarin. The results showed that when the rats were pretreated with verapamil, the maximum concentration (Cmax) of puerarin increased from 683.7 ± 51.2 to 933.5 ± 75.8 ng/mL (p < 0.05), and the area under the concentration-time curve from zero to infinity (AUC0-inf) also increased from 3687.3 ± 444.6 to 5006.1 ± 658.6 µg·h/L (p < 0.05). The Caco-2 cell transwell experiments indicated that verapamil could decrease the efflux ratio of puerarin from 1.90 to 1.19 through inhibiting the activity of P-gp. In conclusion, these results indicated that verapamil could affect the pharmacokinetics of puerarin, possibly by increasing the systemic exposure of puerarin by inhibiting the activity of P-gp.

In vitro Inhibitory Effects of Isofraxidin on Human Liver Cytochrome P450 Enzymes.

Isofraxidin is a Coumarin compound widely distributed in plants, such as the Umbelliferae or Chloranthaceae, and it possesses numerous pharmacological activities. However, whether isofraxidin affects the activity of human liver cytochrome P450 (CYP) enzymes remains unclear. In this study, the inhibitory effects of isofraxidin on the 8 human liver CYP isoforms (i.e., 1A2, 3A4, 2A6, 2E1, 2D6, 2C9, 2C19, and 2C8) were investigated in vitro using human liver microsomes. The results showed that isofraxidin inhibited the activity of CYP1A2, 3A4, and 2E1, with IC50 values of 23.01, 15.49, and 15.98 µmol/L, respectively, but that other CYP isoforms were not affected. Enzyme kinetic studies showed that isofraxidin was not only a noncompetitive inhibitor of CYP3A4 but also a competitive inhibitor of CYP1A2 and 2E1, with Ki values of 7.91, 10.14, and 9.30 µmol/L, respectively. In addition, isofraxidin is a time-dependent inhibitor for CYP3A4 with Kinact/KI value of 0.047/12.33 µmol/L-1min-1. The in vitro studies of isofraxidin with CYP isoforms indicate that isofraxidin has the potential to cause pharmacokinetic drug interactions with other coadministered drugs metabolized by -CYP1A2, 3A4, and 2E1. Further clinical studies are needed to evaluate the significance of this interaction.

The efficacy of RGD modified liposomes loaded with vinorelbine plus tetrandrine in treating resistant brain glioma.

Brain glioma is one of the most common and devastating intracranial malignancies with a high mortality. Chemotherapy for brain glioma is not ideal due to blood brain barrier (BBB) and multidrug resistance (MDR). The objectives of the present study were to develop a kind of RGD (Arg-Gly-Asp) tripeptide modified vinorelbine plus tetrandrine liposomes to achieve BBB transportation, MDR reversion and glioma cell targeting simultaneously. The studies were performed on glioma cells, resistant glioma cells and glioma-bearing mice. Results showed that the constructed liposomes with suitable physicochemical properties could significantly enhance the transport across BBB, obviously accumulate in glioma cells, and exhibit evident capabilities in diminishing brain glioma in mice. Action mechanism studies indicated that the enhanced anticancer efficacy could be attribute to the follows: prolonged elimination half-life (7.093 ± 1.311 h); increased AUC0-24 h (28.92 ± 2.66 mg/L*h); transporting across BBB; enhanced cellular uptake; down-regulation on P-gp (0.49 ± 0.06 fold); inducing apoptosis via activating caspase 8, 9, and 3 (2.40 ± 0.22, 3.57 ± 0.29, and 4.33 ± 0.30 folds, respectively). In conclusion, the RGD modified vinorelbine plus tetrandrine liposomes may offer a promising therapeutic strategy for treatment of brain glioma.

In vitro inhibitory effects of bergenin on human liver cytochrome P450 enzymes.

CONTEXT: Bergenin, isolated from the herb of Bergenia purpurascens (Hook. f. et Thoms.) Engl., has anti-inflammatory, antitussive, and wound healing activities. However, whether bergenin affects the activity of human liver cytochrome P450 (CYP) enzymes remains unclear. MATERIALS AND METHODS: In this study, the inhibitory effects of bergenin (100 µM) on the eight human liver CYP isoforms (i.e., 1A2, 3A4, 2A6, 2E1, 2D6, 2C9, 2C19 and 2C8) were investigated, enzyme kinetics and time-dependent inhibition studies were also performed in vitro using human liver microsomes (HLMs). RESULTS: The results showed that bergenin inhibited the activity of CYP3A4, 2E1 and 2C9, with IC50 values of 14.39, 22.83 and 15.11 µM, respectively, but other CYP isoforms were not affected. Enzyme kinetic studies showed that bergenin was not only a non-competitive inhibitor of CYP3A4, but also a competitive inhibitor of CYP2E1 and 2C9, with Ki values of 7.71, 11.39 and 8.89 µM, respectively. In addition, bergenin is a time-dependent inhibitor for CYP3A4 with Kinact/KI value of 0.025/3.50 µM-1 min-1. DISCUSSION AND CONCLUSIONS: The in vitro studies of bergenin with CYP isoforms indicate that bergenin has the potential to cause pharmacokinetic drug interactions with other co-administered drugs metabolized by CYP3A4, 2E1 and 2C9. Further clinical studies are needed to evaluate the significance of this interaction.

[Construction of cTnC-linker-TnI (P) Genes, Expression of Fusion Protein and Preparation of Lyophilized Protein].

In order to construct and express human cardiac troponin C-linker-troponin I(P) [ cTnC-linker-TnI(P)] fusion protein, detect its activity and prepare lyophilized protein, we searched the CDs of human cTnC and cTnI from GenBank, synthesized cTnC and cTnI(30-110aa) into cloning vector by a short DNA sequence coding for 15 neutral amino acid residues. pCold I-cTnC-linker-TnI(P) was constructed and transformed into E. coli BL21(DE3). Then, cTnC-linker-TnI(P) fusion protein was induced by isopropyl-ß-D-thiogalactopyranoside (IPTG). Soluable expression of cTnC-linker-TnI(P) in prokaryotic system was successfully obtained. The fusion protein was purified by Ni²âº Sepharose 6 Fast Flow affinity chromatography with over 95% purity and prepared into lyophilized protein. The activity of purified cTnC-linker-TnI(P) and its lyophilized protein were detected by Wondfo Finecare™ cTnI Test. Lyophilized protein of cTnC-linker-TnI(P) was stable for 10 or more days at 37 °C and 4 or more months at 25 °C and 4 °C. The expression system established in this research is feasible and efficient. Lyophilized protein is stable enough to be provided as biological raw materials for further research.

Intravoxel incoherent motion diffusion-weighted MRI for the evaluation of early spleen involvement in acute leukemia.

Background: The spleen is a frequent organ of leukemia metastasis. This study aimed to investigate the value of intravoxel incoherent motion (IVIM) diffusion-weighted magnetic resonance imaging (MRI) for assessing pathologic changes in the spleen and identifying early spleen involvement in patients with acute leukemia (AL). Methods: Patients with newly diagnosed AL and healthy controls were recruited between June 2020 and November 2022. All participants underwent abdominal IVIM diffusion-weighted imaging (DWI) at our hospital. IVIM parameters [pure diffusion coefficient (D); pseudo-diffusion coefficient (D*); and pseudo-perfusion fraction (f)] of the spleen were calculated by the segmented fitting method, and perfusion-diffusion ratio (PDR) was further calculated from the values of D, D* and f. Spleen volumes (SVs) were obtained by manually segmenting the spleen layer by layer. Clinical biomarkers of AL patients were collected. Patients were divided into splenomegaly group and normal SV group according to the individualized reference intervals for SV. IVIM parameters were compared among the control group, AL with normal SV group, and AL with splenomegaly group using one-way analysis of variance, followed by pairwise post hoc comparisons. The correlations of IVIM parameters with clinical biomarkers were analyzed in AL patients. The diagnostic performances of IVIM parameters and their combinations for differentiating among the three groups were compared. Results: Seventy-nine AL patients (AL with splenomegaly: n=54; AL with normal SV: n=25) and 55 healthy controls were evaluated. IVIM parameters were significantly different among the three groups (P<0.001 for D, D* and f; P=0.001 for PDR). D and PDR showed significant differences between the control and AL with normal SV groups in pairwise comparisons (P<0.001, and P=0.031, respectively). D was correlated with white blood cell (WBC) counts (r=-0.424; 95% CI: -0.570, -0.211; P<0.001), lactate dehydrogenase (LDH) (r=-0.285; 95% CI: -0.486, -0.011; P=0.011), and bone marrow blasts (r=-0.283; 95% CI: -0.476, -0.067; P=0.012). D* (r=-0.276; 95% CI: -0.470, -0.025; P=0.014), f (r=0.514; 95% CI: 0.342, 0.664; P<0.001) and PDR (r=0.343; 95% CI: 0.208, 0.549; P=0.002) were correlated with LDH. The combination of IVIM parameters (AUC: 0.830; 95% CI: 0.729, 0.905) demonstrated better diagnostic efficacy than the single D* (AUC: 0.721; 95% CI: 0.608, 0.816; Delong test: Z=2.012, P=0.044) and f (AUC: 0.647; 95% CI: 0.532, 0.752; Delong test: Z=2.829, P=0.005), but was not significantly different from the single D (AUC: 0.756; 95% CI: 0.647, 0.846; Delong test: Z=1.676, P=0.094) in differentiating the splenomegaly group and normal SV group. Conclusions: IVIM diffusion-weighted MRI could be a potential alternative for assessing pathologic changes in the spleen from cellularity and angiogenesis, and D and PDR may be viable indicators to identify early spleen involvement in patients with AL.