Purification and Structural Analyses of Sulfated Polysaccharides from Low-Value Sea Cucumber Stichopus naso and Anticoagulant Activities of Its Oligosaccharides.

Three polysaccharides (SnNG, SnFS and SnFG) were purified from the body wall of Stichopus naso. The physicochemical properties, including monosaccharide composition, molecular weight, sulfate content, and optical rotation, were analyzed, confirming that SnFS and SnFG are sulfated polysaccharides commonly found in sea cucumbers. The highly regular structure {3)-L-Fuc2S-(α1,}n of SnFS was determined via a detailed NMR analysis of its oxidative degradation product. By employing ß-elimination depolymerization of SnFG, tri-, penta-, octa-, hendeca-, tetradeca-, and heptadeca-saccharides were obtained from the low-molecular-weight product. Their well-defined structures confirmed that SnFG possessed the backbone of {D-GalNAc4S6S-ß(1,4)-D-GlcA}, and each GlcA residue was branched with Fuc2S4S. SnFS and SnFG are both structurally the simplest version of natural fucan sulfate and fucosylated glycosaminoglycan, facilitating the application of low-value sea cucumbers S. naso. Bioactivity assays showed that SnFG and its derived oligosaccharides exhibited potent anticoagulation and intrinsic factor Xase (iXase) inhibition. Moreover, a comparative analysis with the series of oligosaccharides solely branched with Fuc3S4S showed that in oligosaccharides with lower degrees of polymerization, such as octasaccharides, Fuc2S4S led to a greater increase in APTT prolongation and iXase inhibition. As the degree of polymerization increases, the influence from the sulfation pattern diminishes, until it is overshadowed by the effects of molecular weight.

Breast Magnetic Resonance Imaging Can Predict Ki67 Discordance Between Core Needle Biopsy and Surgical Samples.

BACKGROUND: Disagreement in assessments of Ki67 expression based on core-needle biopsy and matched surgical samples complicates decisions in the treatment of breast cancer. PURPOSE: To examine whether preoperative breast MRI could be useful in predicting Ki67 discordance between core-needle biopsy and surgical samples. STUDY TYPE: Retrospective. POPULATION: Three hundred and sixty-five breast cancer patients with MRI scans and having both core-needle biopsy and surgical samples from 2017 to 2019. FIELD STRENGTH/SEQUENCE: 3.0 T, T2-weighted iterative decomposition of water and fat with echo asymmetry and least squares estimation sequence, diffusion-weighted sequence using b-values 0/1000, dynamic contrast enhanced image by volume imaged breast assessme NT. ASSESSMENT: We collected clinicopathologic variables and preoperative MRI features (tumor size, lesion type, shape of mass, spiculated margin, internal enhancement, peri-tumoral edema, intra-tumoral necrosis, multifocal/multicentric, apparent diffusion coefficient [ADC] minimum, ADC mean, ADC maximum, ADC difference). STATISTICAL TESTS: K-means clustering, multivariable logistic regression, receiver operating characteristic curve. RESULTS: Sixty-one patients showed Ki67 discordance and 304 patients show Ki67 concordance according to our definition using K-means clustering. Multivariable regression analysis showed that the following parameters were independently associated with Ki67 discordance: peri-tumoral edema, odds ratio (OR) 2.662, 95% confidence interval (CI) 1.432-4.948; ADCmin ≤ 0.829 × 10-3  mm2 /sec, OR 2.180, 95% CI 1.075-4.418; and ADCdiff > 0.317 × 10-3  mm2 /sec, OR 3.365, 95% CI 1.698-6.669. This multivariable model resulted in an AUC of 0.758 (95% CI 0.711-0.802) with sensitivity and specificity being 0.803 and 0.621, respectively. CONCLUSION: Presence of peri-tumoral edema, smaller ADCmin and greater ADCdiff in preoperative breast MRI may indicate high risk of Ki67 discordance between core-needle biopsy and surgical samples. For patients with these MRI-based risk factors, clinicians should not rely on Ki67 assessment only from core-needle biopsy.

A functional liver imaging score for preoperative prediction of liver failure after hepatocellular carcinoma resection.

OBJECTIVES: Posthepatectomy liver failure (PHLF) is a challenging complication after resection to treat hepatocellular carcinoma (HCC), and it is associated with high mortality. Preoperative prediction of PHLF may improve patient subsequent and reduce such mortality. This study examined whether a functional liver imaging score (FLIS) based on preoperative gadoxetic acid-enhanced magnetic resonance imaging (MRI) could predict PHLF. MATERIALS AND METHODS: The study included 502 patients who underwent preoperative gadoxetic acid-enhanced MRI, followed by HCC resection. Significant preoperative predictors of PHLF were identified using logistic regression analysis. The ability of FLIS to predict PHLF was evaluated using receiver operating characteristic curves, and its predictive power was compared to that of the model for end-stage liver disease (MELD) score, albumin-bilirubin (ALBI) score, and indocyanine green 15-min retention rate (ICG-R15). RESULTS: In multivariate analysis, PHLF was independently associated with FLIS (OR 0.452, 95% CI 0.361 to 0.568, p < 0.001) and major resection (OR 1.898, 95% CI 1.057 to 3.408, p = 0.032). FLIS was associated with a higher area under the receiver operating characteristic curve (0.752) than the MELD score (0.557), ALBI score (0.609), or ICG-R15 (0.605) (all p < 0.05). Patients with FLIS ≤ 4 who underwent major resection were at 9.4-fold higher risk of PHLF than patients with lower FLIS who underwent minor resection. CONCLUSION: FLIS is an independent predictor of PHLF, and it may perform better than the MELD score, ALBI score, and ICG-R15 clearance. We propose treating elevated FLIS and major resection as risk factors for PHLF. KEY POINTS: • A functional liver imaging score can independently predict posthepatectomy liver failure in patients with HCC. • The score may predict such failure better than MELD and ALBI scores and ICG-R15. • Patients with scores ≤ 4 who undergo major hepatic resection may be at nearly tenfold higher risk of posthepatectomy liver failure.

Preoperative normalized iodine concentration derived from spectral CT is correlated with early recurrence of hepatocellular carcinoma after curative resection.

OBJECTIVES: To investigate whether normalized iodine concentration (NIC) correlates with tumor microvessel density and early recurrence in patients with HCC. MATERIALS AND METHODS: We included 71 patients with surgically resected single HCC in this prospective study who underwent preoperative spectral CT between November 2014 and June 2016. Two observers independently measured the NIC in the arterial phase (AP) and portal venous phase (PVP). The relationship between NIC and microvessel density was evaluated. Univariate and multivariate logistic regression was performed to evaluate independent predictors of early recurrence. RESULTS: Early recurrence occurred in 28 of 71 patients (39.4%) during the 2-year follow-up. NIC-AP positively correlated with microvessel density for the two observers (r = 0.593 and 0.527). Based on multivariate analysis, independent risk factors for early HCC recurrence were tumor size (odds ratio, 1.200; p = 0.043) and NIC-AP (odds ratio, 2.522; p = 0.005). For the two observers, areas under the receiver operating characteristic curve for predicting early HCC recurrence were 0.719 and 0.677. Early recurrence rates were significantly higher among patients with NIC-AP values higher than the optimal cutoff than among those with values below the cutoff. CONCLUSION: Normalized iodine concentration in the arterial phase from spectral CT reflects tumor-derived angiogenesis and is a potential predictive biomarker for early recurrence of hepatocellular carcinoma. KEY POINTS: • Normalized iodine concentration in the arterial phase positively correlated with microvessel density of hepatocellular carcinoma. • In the patients with hepatocellular carcinoma, tumor size and normalized iodine concentration in the arterial phase were independent risk factors for early hepatocellular carcinoma recurrence. • Early hepatocellular carcinoma recurrence rates were significantly higher when normalized iodine concentration in the arterial phase values was above the optimal cutoff.

Value of Diffusion-Weighted Imaging Combined with Susceptibility-Weighted Imaging in Differentiating Benign from Malignant Parotid Gland Lesions.

BACKGROUND The aim of this study was to investigate the diagnostic value of diffusion-weighted imaging (DWI) in combination with susceptibility-weighted imaging (SWI) for differentiating benign parotid gland lesions from malignant ones. MATERIAL AND METHODS This retrospective study was approved by the Ethics Committee of our hospital. A total of 36 patients (26 benign cases and 10 malignant cases) were confirmed by surgical pathology. The apparent diffusion coefficient (ADC), normalized ADC (ADCNormalized), intratumoral susceptibility signals (ITSS), and morphological characteristics were analyzed with SPSS 19.0 software. RESULTS The mean ADC values of parotid gland lesions was not different between malignant and benign lesions (P=0.07), while the differences between ADCNormalized (P=0.026) and ITSS grading (P=0.014) were statistically significant. Logistic regression analysis identified use of ADCNormalized and ITSS as the only independent predictor of malignant lesions (odds ratio 0.038; 95% confidence interval 0.001~0.988; P=0.011) and (odds ratio 4.867; 95% confidence interval 1.442~16.423; P=0.049), respectively. The optimum threshold of the ADCNormalized values was -0.45%, ITSS grade was 2, the corresponding areas under the receiver operating characteristic curve (AUC) were 0.750 and 0.787 respectively, and the combination of the 2 was 0.846. CONCLUSIONS DWI integrated with SWI can significantly improve the diagnostic efficacy in distinguishing benign from malignant parotid lesions.

GSTT1 and GSTM1 polymorphisms predict treatment outcome for breast cancer: a systematic review and meta-analysis.

Observational studies have reported controversial results on the association between GSTT1 and GSTM1 genotypes and treatment outcome of breast cancer. The purpose of this study is to evaluate the association between GSTT1 and GSTM1 and treatment outcome in breast cancer patients. Eligible studies were searched in PubMed, EMBASE, Cochrane Library, and China National Knowledge Infrastructure databases. A random-effect model or fixed-effect model was used to calculate the overall combined risk estimates. Twenty-one studies with a total of 4990 patients were included in this meta-analysis. The GSTM1 null genotype (odds ratio (OR) = 1.33, 95 % confidence interval (CI) 1.01-1.75, P = 0.046) and GSTT1/GSTM1 double null genotype (OR = 2.22, 95 % CI 1.02-4.84, P = 0.045) were significantly associated with an increased tumor response. A reduced overall survival (hazard ratio (HR) = 0.84, 95 % CI 0.72-0.98, P = 0.024) was observed in GSTM1 null genotype, especially in mixed descent (HR = 0.77, 95 % CI 0.61-0.96, P = 0.018) and large sample size (HR = 0.85, 95 % CI 0.72-0.99, P = 0.033). Evidence of publication bias was observed in GSTM1 genotype rather than in GSTT1 genotype. This meta-analysis suggests that GSTM1 null and GSTT1/GSTM1 double null polymorphisms might be significantly associated with an increased tumor response. However, the GSTM1 null genotype might be significantly associated with a reduced overall survival. Future studies are warranted to confirm these findings.

Regulation of histone acetyltransferase TIP60 function by histone deacetylase 3.

The key member of the MOZ (monocyticleukaemia zinc finger protein), Ybf2/Sas3, Sas2, and TIP60 acetyltransferases family, Tat-interactive protein, 60 kD (TIP60), tightly modulates a wide array of cellular processes, including chromatin remodeling, gene transcription, apoptosis, DNA repair, and cell cycle arrest. The function of TIP60 can be regulated by SIRT1 through deacetylation. Here we found that TIP60 can also be functionally regulated by HDAC3. We identified six lysine residues as its autoacetylation sites. Mutagenesis of these lysines to arginines completely abolishes the autoacetylation of TIP60. Overexpression of HDAC3 increases TIP60 ubiquitination levels. However, unlike SIRT1, HDAC3 increased the half-life of TIP60. Further study found that HDAC3 colocalized with TIP60 both in the nucleus and the cytoplasm, which could be the reason why HDAC3 can stabilize TIP60. The deacetylation of TIP60 by both SIRT1 and HDAC3 reduces apoptosis induced by DNA damage. Knockdown of HDAC3 in cells increased TIP60 acetylation levels and increased apoptosis after DNA damage. Together, our findings provide a better understanding of TIP60 regulation mechanisms, which is a significant basis for further studies of its cellular functions.

Monitoring of Chlorophyll Content of Potato in Northern Shaanxi Based on Different Spectral Parameters.

Leaf chlorophyll content (LCC) is an important physiological index to evaluate the photosynthetic capacity and growth health of crops. In this investigation, the focus was placed on the chlorophyll content per unit of leaf area (LCCA) and the chlorophyll content per unit of fresh weight (LCCW) during the tuber formation phase of potatoes in Northern Shaanxi. Ground-based hyperspectral data were acquired for this purpose to formulate the vegetation index. The correlation coefficient method was used to obtain the "trilateral" parameters with the best correlation between potato LCCA and LCCW, empirical vegetation index, any two-band vegetation index constructed after 0-2 fractional differential transformation (step size 0.5), and the parameters with the highest correlation among the three spectral parameters, which were divided into four combinations as model inputs. The prediction models of potato LCCA and LCCW were constructed using the support vector machine (SVM), random forest (RF) and back propagation neural network (BPNN) algorithms. The results showed that, compared with the "trilateral" parameter and the empirical vegetation index, the spectral index constructed by the hyperspectral reflectance after differential transformation had a stronger correlation with potato LCCA and LCCW. Compared with no treatment, the correlation between spectral index and potato LCC and the prediction accuracy of the model showed a trend of decreasing after initial growth with the increase in differential order. The highest correlation index after 0-2 order differential treatment is DI, and the maximum correlation coefficients are 0.787, 0.798, 0.792, 0.788 and 0.756, respectively. The maximum value of the spectral index correlation coefficient after each order differential treatment corresponds to the red edge or near-infrared band. A comprehensive comparison shows that in the LCCA and LCCW estimation models, the RF model has the highest accuracy when combination 3 is used as the input variable. Therefore, it is more recommended to use the LCCA to estimate the chlorophyll content of crop leaves in the agricultural practices of the potato industry. The results of this study can enhance the scientific understanding and accurate simulation of potato canopy spectral information, provide a theoretical basis for the remote sensing inversion of crop growth, and promote the development of modern precision agriculture.

Mitochondrial Lon protease promotes CD4+ T cell activation by activating the cGAS-STING-TBK1 axis in systemic lupus erythematosus.

Systemic lupus erythematosus (SLE) is an autoimmune disease in which autoreactive CD4+ T cells play an essential role. We extracted CD4+ T cells from SLE-prone Fcgr2b-/- mice to elaborate the mechanism of mitochondrial Lon protease in CD4+ T cell activation in SLE. Transcriptome sequencing was performed in SLE-prone Fcgr2b-/- mice, and the stimulator of interferon gene (STING) related to SLE was obtained. It was demonstrated that STING expression was elevated in CD4+ T cells in SLE-prone Fcgr2b-/- mice. The downstream genes and pathways of STING were predicted by GO and KEGG approaches. The data indicated that STING regulated IFN signaling to promote CD4+ T cell activation in SLE-prone Fcgr2b-/- mice. Next, the interaction of cGAS, STING, TBK1, and IFN-I was verified by Co-IP assay. Moreover, the roles of cGAS, STING, and TBK1 in activating CD4+ T cells from SLE-prone Fcgr2b-/- mice were evaluated using gain- or loss-of-function experiments. Mechanistically, cGAS upregulated the IFN-I signaling pathway by directly interacting with STING and TBK1, contributing to CD4+ T cell activation. Besides, cytosolic mtDNA could activate CD4+ T cell activation in SLE-prone Fcgr2b-/- mice by upregulating the cGAS-STING-TBK1 axis. The function of mitochondrial Lon protease in oxidative damage and mtDNA release in CD4+ T cells of SLE-prone Fcgr2b-/- mice were explored. Mitochondrial Lon protease enhanced mtDNA release into the cytoplasm under oxidative stress. Collectively, our work indicates that mitochondrial Lon protease enhances CD4+ T cell activation by inducing mtDNA leakage and offers new candidate targets for developing diagnostic and therapeutic strategies.

A Reliable and Repeatable Model for Predicting Microvascular Invasion in Patients With Hepatocellular Carcinoma.

RATIONALE AND OBJECTIVES: The reproducibility of imaging models for predicting microvascular invasion (MVI) in patients with hepatocellular carcinoma (HCC) remains questionable due to inconsistent interpretation of image signs. Our aim was to screen for high-consensus MRI features to develop a repeatable model for predicting MVI. MATERIALS AND METHODS: We included 219 patients with HCC who underwent surgical resection, and patients were divided into a training cohort (n = 145) and a validation cohort (n = 74). Morphological characteristics, signal features on hepatobiliary phases, and dynamic enhancement patterns were qualitatively interobserver evaluated. Interobserver agreement was assessed using Cohen's κ for selecting features with high interobserver agreement. Risk factors that were significant in stepwise multivariate analysis and that could be measured with good interobserver agreement were used to construct a predictive model, which was assessed in the validation cohort. The diagnostic performance of the model was evaluated based on area under the receiver operating characteristic curve (AUC). RESULTS: Multivariate analysis identified nonsmooth tumor margin, absence of radiologic capsule, and intratumoral artery as independent risk factors of MVI. These MRI-based features showed good or nearly perfect interobserver agreement between radiologists (κ > 0.6). The predictive model predicted MVI well in the training (AUC 0.734) and validation cohorts (AUC 0.759) and fitted well to calibration curves. CONCLUSION: MRI features included nonsmooth tumor margin, absence of radiologic capsule, and intratumoral artery that can be assessed with high interobserver agreement can predict MVI in HCC patients. The predictive model described here may be useful to radiologists, regardless of experience level.

Efficacy and Safety of CMAB008 Compared with Innovator Infliximab in Patients with Moderate-to-Severe Rheumatoid Arthritis Receiving Concomitant Methotrexate: A Randomized, Double-blind, Multi-center, Phase III Non-inferiority Study.

OBJECTIVES: The aim of this work is to verify the non-inferior efficacy and safety of CMAB008 compared with innovator infliximab in rheumatoid arthritis patients combined with methotrexate. METHODS: We conducted a randomized, double-blinded, parallel, positive control design, multicenter study, with a stable dose of methotrexate. Patients were enrolled randomly with a ratio of 1:1 to receive intravenously CMAB008 3 mg/kg or innovator infliximab 3 mg/kg at weeks 0, 2, 6, 14, 22 and 30. The primary efficacy endpoint was American College of Rheumatology 20% improvement criteria (ACR20) response rate at week 30. The non-inferiority was established if the lower limit of the one-sided 97.5% confidence interval (CI) for the difference was more than - 15% and the equivalence was established if the two-sided 95% CI was within ± 15% in an exploratory equivalence analysis. The secondary endpoints included other efficacy assessment parameters, as well as immunogenicity, safety, and pharmacokinetics. RESULTS: In the full analysis population (FAS), 110 (57.6%) of 191 patients in the CMAB008 group and 120 (62.2%) of 193 patients in the innovator infliximab group reached the primary outcome of ACR20 at week 30. The differences of the rates were - 4.6% and the lower limit of one-sided 97.5% confidence interval was - 14.29%, not less than the lower limit of the non-inferiority margin (- 15%); so CMAB008 was non-inferior to innovator infliximab. Further, CMAB008 was equivalent to innovator infliximab both in FAS (difference - 4.6%, 95% CI - 14.29% to 5.12%) and PPS (difference - 3.3%, 95% CI - 13.18% to 6.62%). The efficacy, safety, immunogenicity, and pharmacokinetics are highly similar between CMAB008 and innovator infliximab. CONCLUSIONS: Non-inferior efficacy of CMAB008 to innovator infliximab is illustrated with similar early and lasting therapeutic effects, and the equivalence is further demonstrated. CMAB008 is well tolerated and has semblable safety compared with the innovator infliximab. TRIAL REGISTRATION NUMBER: NCT03478111.

Pegmolesatide for the treatment of anemia in patients undergoing dialysis: a randomized clinical trial.

Background: Pegmolesatide, a synthetic peptide-based erythropoietin (EPO) receptor agonist, is being evaluated as an alternative to epoetin alfa for treating anemia of chronic kidney disease (CKD) in Chinese dialysis patients. There is a critical need for a long-acting, cost-effective erythropoiesis-stimulating agent that does not produce EPO antibodies. Methods: A randomized, open-label, active-comparator, non-inferiority phase three trial was conducted at 43 dialysis centers in China between May 17th, 2019, and March 28th, 2022. Eligible patients aged 18-70 years were randomly assigned (2:1) to receive pegmolesatide once every four weeks or epoetin alfa one to three times per week, with doses adjusted to maintain a hemoglobin level between 10.0 and 12.0 g/dL. The primary efficacy endpoint was the mean change in hemoglobin level from baseline to the efficacy evaluation period in the per-protocol set (PPS) population. Non-inferiority of pegmolesatide to epoetin alfa was established if the lower limit of the two-sided 95% confidence interval for the between-group difference was ≥ -1.0 g/dL. Safety assessment included adverse events and potential anaphylaxis reactions. This trial is registered at ClinicalTrials.gov, NCT03902691. Findings: Three hundreds and seventy-two patients were randomly assigned to the pegmolesatide group (248 patients) or the epoetin alfa group (124 patients). A total of 347 patients (233 in the pegmolesatide group and 114 in the epoetin alfa group) were included in the PPS population. In the PPS, the mean change (standard deviation, SD) in hemoglobin level from baseline to the efficacy evaluation period was 0.07 (0.92) g/dL in the pegmolesatide group and -0.22 (0.97) g/dL in the epoetin alfa group. The between-group difference was 0.29 g/dL (95% confidence interval: 0.11-0.47), verifying non-inferiority of pegmolesatide to epoetin alfa. Adverse events occurred in 231 (94%) participants in the pegmolesatide group and in 110 (89%) in the epoetin alfa group. Hypertension was the most common treatment-related adverse event. No fatal cases of anaphylaxis or hypotension were reported. Interpretation: Monthly subcutaneously injection of pegmolesatide was as effective and safe as conventional epoetin alfa administrated one to three times a week in treating anemia in Chinese dialysis patients. Funding: The study was supported by Hansoh Medical Development Group.

[Peripheral blood leukocyte double strand RNA-dependent protein kinase gene expression in patients with systemic lupus erythematosus].

OBJECTIVE: To investigate the expression of the double-stranded RNA-dependent protein kinase (PKR) gene in the peripheral blood leukocyte of patients with systemic lupus erythematosus (SLE), and to evaluate the relationship between the gene expression and the disease activity. METHODS: The clinical data of 100 SLE patients, 40 non-SLE patients with rheumatic diseases, and 40 normal controls were collected. Total RNA was extracted from the peripheral blood and then reverse transcribed into cDNA. Sybr green dye based real-time quantitative PCR method was used to compare the expression levels (indicated as 2(-ΔCt) value) of PKR in the three groups. RESULTS: (1) The 2(-ΔCt) value of PKR expression level in the SLE patients was (14.69 ± 7.62), which was significantly higher than those in the non-SLE patients (5.09 ± 4.73, P = 0.012)and normal controls(4.79 ± 3.49, P = 0.005). (2) The 2(-ΔCt) value of PKR expression level in the SLE patients with severe activity was (22.57 ± 2.61), which was significantly higher than those in the SLE patients with mild activity and no activity (12.94 ± 2.41, P = 0.000; 8.85 ± 2.17, P = 0.000). (3) The 2(-ΔCt) value of PKR expression level in the SLE patients with lupus nephritis was significantly higher than that in the SLE patients without lupus nephritis (16.85 ± 7.32 vs 8.35 ± 2.04, P = 0.034). (4) The 2(-ΔCt) value of PKR was correlated with the systemic lupus erythematosus index (SLEDAI) scores (r = 0.32, P = 0.000), WBC (r = 0.46, P = 0.000), Hb (r = -0.22, P = 0.035), the quantitation of urine protein in 24 hours (r = 0.21, P = 0.000), HDL-C (r = 0.21, P = 0.022), and anti-RNP antibody (r(s) = -0.21, P = 0.025). CONCLUSIONS: The expression of PKR in the SLE patients is up-regulated, especially in those with severe activity. The expression level of PKR gene is associated with SLE disease activity.

Ultrasound and magnetic resonance imaging of cyclic arginine glycine aspartic acid-gadopentetic acid-polylactic acid in human breast cancer by targeting αvß3 in xenograft-bearing nude mice.

Effective early detection shows the potential to reduce breast cancer mortality. This study aimed to establish a targeted contrast agent for Magnetic Resonance Imaging (MRI)/ultrasound dual-modality molecular radiography for breast cancer. The cyclic arginine-glycine-aspartate-gadopentetic acid-polylactic acid (cRGD and Gd-DTPA) coated by multi-functional blank poly (lactic-co-glycolic acid) (PLGA) nanoparticles) was successfully constructed by chemical synthesis method with high stability. The safety of cRGD-Gd-DTPA-PLGA was demonstrated in vitro and in vivo, and their affinity to breast cancer cells was revealed. Moreover, MRI/ultrasound dual-modality molecular radiography in vitro showed that as the concentration of contrast agent increased, the echo enhancement and signal intensity of MRI imaging were also elevated. The mouse models of human breast cancer also indicated significant target enhancements of cRGD-Gd-DTPA-PLGA magnetic nanoparticles in the mouse tumor. Thus, cRGD-Gd-DTPA-PLGA magnetic nanoparticles were suggested as qualified MRI/ultrasound dual-modality molecular radiography contrast agent. We further explored the targeting mechanism of cRGD-Gd-DTPA-PLGA in breast cancer. The results showed that αvß3 was highly expressed in breast cancer tissues, and cRGD-Gd-DTPA-PLGA used for MRI/ultrasound dual-modality molecular radiography by targeting αvß3. Additionally, we found that the signal-to-noise ratio of MRI was positively correlated with microvessel density (MVD). The cRGD-Gd-DTPA-PLGA dynamicly and quantitatively monitored breast cancer by monitoring the state of neovascularization. In conclusion, in the present study, we successfully constructed the cRGD-Gd-DTPA-PLGA magnetic nanoparticles for MRI/ultrasound dual-modality molecular radiography. The cRGD-Gd-DTPA-PLGA showed potential in early detection and diagnosis of metastasis, and dynamic evaluation of the efficacy of molecular targeted therapy of integrin αvß3.

Comparison of the efficacy and safety of the adalimumab biosimilar TQ-Z2301 and adalimumab for the treatment of Chinese patients with active ankylosing spondylitis: a multi-center, randomized, double-blind, phase III clinical trial.

OBJECTIVE: To assess the clinical equivalence of TQ-Z2301, a biosimilar of adalimumab, to the reference adalimumab in the treatment of Chinese patients with active ankylosing spondylitis. METHODS: This multicenter, randomized, double-blind, positive-controlled phase III clinical trial was conducted in 19 centers across China. Chinese adults with active ankylosing spondylitis despite being treated with non-steroidal anti-inflammatory drugs for ≥ 4 weeks were randomized in a 1:1 ratio to subcutaneously receive 40 mg of TQ-Z2301 or adalimumab every other week for 24 weeks. The primary endpoint was the percentage of patients who achieved at least 20% improvement according to the Assessment of Spondyloarthritis International Society criteria (ASAS20) at week 24. The equivalence was established if the 90% CI for RR of ASAS20 between two groups at week 24 fell within (0.80, 1.25). Secondary endpoints included efficacy measures of disease activity, spinal mobility, physical function and quality of life, immunogenicity, and pharmacokinetic parameters. Safety analysis was done for all patients who received at least one study drug. RESULTS: A total of 380 patients were enrolled in the study between September 2018 and October 2019, including 188 in the TQ-Z2301 group and 192 in the adalimumab group. In the full analysis population, the ASAS20 response rate at week 24 was 86.70% in the TQ-Z2301 group, and 80.73% in the adalimumab group, the RR of ASAS20 for TQ-Z2301 versus adalimumab was 1.074, 90% CI (0.997, 1.157), fell within the predefined equivalence boundary (0.80, 1.25). Except for the SF-36 at week 12, there was no statistical difference between the two groups for all the secondary endpoints (P>0.05). The incidence of adverse events group was 82.45% in the TQ-Z2301, and 83.85% in the adalimumab group, the safety profile of the two groups was similar. The profiles of immunogenicity and pharmacokinetics were also similar between the two groups. CONCLUSION: TQ-Z2301 is equivalent to adalimumab for the treatment of Chinese patients with active ankylosing spondylitis. The safety, immunogenicity, and pharmacokinetic characteristics of both drugs are similar. TRIAL REGISTRATION: The study (CTR20181863) was registered in the Chinese Clinical Trial Registry on 19 October 2018. Key Points • TQ-Z2301 showed the equivalence of efficacy compared with the reference adalimumab for the treatment of Chinese patients with active ankylosing spondylitis. • The safety, immunogenicity, and pharmacokinetics profiles of TQZ-2301 were similar to those of the reference adalimumab.

Effect of Tacrolimus vs Intravenous Cyclophosphamide on Complete or Partial Response in Patients With Lupus Nephritis: A Randomized Clinical Trial.

Importance: Lupus nephritis (LN) is typically treated with intravenous cyclophosphamide (IVCY), which is associated with serious adverse effects. Tacrolimus may be an alternative for initial treatment of LN; however, large-scale, randomized clinical studies of tacrolimus are lacking. Objective: To assess efficacy and safety of tacrolimus vs IVCY as an initial therapy for LN in China. Design, Setting, and Participants: This randomized (1:1), open-label, parallel-controlled, phase 3, noninferiority clinical trial recruited patients aged 18 to 60 years with systemic lupus erythematosus and LN class III, IV, V, III+V, or IV+V primarily from outpatient settings at 35 centers in China. Inclusion criteria included body mass index of 18.5 or greater to less than 27, 24-hour urine protein of 1.5 g or greater, and serum creatinine of less than 260 µmol/L. Of 505 patients screened, 191 failed screening (163 ineligible, 25 withdrawn consent, and 3 other reasons). Overall, 314 were randomized. The first patient was enrolled March 10, 2015, and the study finished September 13, 2018. The follow-up period was 24 weeks. Data were analyzed from December 2019 to March 2020. Interventions: Oral tacrolimus (target trough level, 4-10 ng/mL) or IVCY for 24 weeks plus prednisone. Main Outcomes and Measures: Complete or partial response rate at week 24 (prespecified). Results: A total of 314 patients were randomized (158 [50.3%] to tacrolimus and 156 [49.7%] to IVCY). Overall, 299 patients (95.2%) were treated (tacrolimus group, 157 [52.5%]; IVCY group, 142 [47.5%]). Baseline demographic and clinical characteristics were generally similar between groups (mean [SD] age, 34.2 [9.5] years; 262 [87.6%] female). Tacrolimus was found to be noninferior to IVCY for LN response at week 24. There was a complete or partial response rate of 83.0% (117 of 141 patients) in the tacrolimus group and 75.0% (93 of 124 patients) in the IVCY group (difference, 7.1%; 2-sided 95% CI, -2.7% to 16.9%; lower limit of 95% CI greater than -15%). At week 24, least-square mean change in Systemic Lupus Erythematosus Disease Activity Index score was -8.6 with tacrolimus and -6.4 with IVCY (difference, -2.2; 95% CI, -3.1 to -1.3). Changes in other immune parameters and kidney function were generally similar between groups. Serious treatment-emergent adverse events (TEAEs) were reported in 29 patients in the tacrolimus group (18.5%) and 35 patients in the IVCY group (24.6%). Most common serious study drug-related TEAEs were infections (14 [8.9%] and 23 [16.2%], respectively). Seven patients in each group withdrew due to AEs. Conclusions and Relevance: In this study, oral tacrolimus appeared noninferior to IVCY for initial therapy of active LN, with a more favorable safety profile than IVCY. Tacrolimus may be an alternative to IVCY as initial therapy for LN. Trial Registration: ClinicalTrials.gov Identifier: NCT02457221.

Glycogen synthase kinase 3 drives thymocyte egress by suppressing ß-catenin activation of Akt.

Molecular pathways controlling emigration of mature thymocytes from thymus to the periphery remain incompletely understood. Here, we show that T cell­specific ablation of glycogen synthase kinase 3 (GSK3) led to severely impaired thymic egress. In the absence of GSK3, ß-catenin accumulated in the cytoplasm, where it associated with and activated Akt, leading to phosphorylation and degradation of Foxo1 and downregulation of Klf2 and S1P1 expression, thereby preventing emigration of thymocytes. A cytoplasmic membrane-localized ß-catenin excluded from the nucleus promoted Akt activation, suggesting a new function of ß-catenin independent of its role as a transcriptional activator. Furthermore, genetic ablation of ß-catenin, retroviral expression of a dominant negative Akt mutant, and transgenic expression of a constitutively active Foxo1 restored emigration of GSK3-deficient thymocytes. Our findings establish an essential role for GSK3 in thymocyte egress and reveal a previously unidentified signaling function of ß-catenin in the cytoplasm.

[Effect and mechanism of LY294002 on growth of fibrosarcoma cell line HT1080].

OBJECTIVE: To investigate the effect and mechanism of LY294002 on growth of fibrosarcoma cell line HT1080. METHODS: The proliferation inhibitory rate of HT1080 cells treated by LY294002 at doses of 5, 10, 25, 50, 100 micromol/L for 12 and 24 h, respectively, was evaluated using SunBio Am-Blue method. HT1080 cells were divided into two groups, that is, A and B. The A group was control group without treatment. The B group received LY294002 (100 micromol/L) for 24 h. The changes of cell morphology and quantity were observed by phase contrast microscope. The apoptosis rate of HT1080 cells was detected by flow cytometry. And the protein expression of p-Akt and p-mTOR in HT1080 cells were detected by Western Blotting. RESULTS: The proliferation of HT1080 cells was inhibited in time- and dose- dependent manner by LY294002. After the treatment of 100 micromol/L LY294002 for 24 h, the growth of HT1080 cell line was remarkably inhibited by LY294002. The rate of apoptosis increased. And the protein expression of p-Akt (0.23 +/- 0.01) and p-mTOR (0.32 +/- 0.06) in LY294002 group was lower than p-Akt (0.63 +/- 0.02) and p-mTOR (0.71 +/- 0.02) in control group (P<0. 01). CONCLUSION: LY294002 can inhibit the growth of HT1080 cells through PI3K-mTOR pathway. PI3K-mTOR pathway presents an appealing therapeutic target on fibrosarcoma.

[Interferon-inducible genes 2', 5'-ollgoadenylate synthetase-like and tetratricopeptide repeats 2 are correlated with clinical features of patients with systemic lupus erythematosus].

OBJECTIVE: To investigate the expression levels of 2', 5'-oligoadenylate synthetase-like (OASL) and interferon induced protein with tetratricopeptide repeats 2 (IFIT2) genes in the peripheral blood leukocyte of patients with systemic lupus erythematosus (SLE), and to evaluate the relations between these gene expression levels and disease activity. METHODS: The clinical data of 50 SLE patients, 25 non-SLE patients with rheumatic diseases, and 25 normal controls were collected. Specimens of peripheral blood were collected; total RNA was extracted and transcribed into cDNA. SYBR green dye based real-time quantitative PCR method was applied to compare the expression levels (indicated as deltaCT value) of OASL and IFIT2 in patients with SLE and those in the controls. RESULTS: The deltaCT value of OASL expression level of the SLE patients was (4.83 +/- 0.41), significantly higher than those of the non-SLE patients (3.26 +/- 0.47) and normal controls (3.07 +/- 0.54, both P < 0.05), The deltaCT value of IFIT2 expression level of the SLE patients was (2.85 +/- 0.41), significantly higher than those of the non-SLE patients (1.19 +/- 0.52) and normal controls (1.07 +/- 0.47, both P < 0.05). The deltaCT value of OASL is related with the SLEDAI scores and immunoglobulin A. (Both P < 0.05). The deltaCT value of IFIT2 is related with the SLEDAI scores and IgA, white blood cell (both P < 0.05). CONCLUSION: The value of IFIT2 and OASL expression level of the SLE patients is up-regulation, the real time expression levels of OASL and IFIT2 genes are associated with SLE disease activity. To inhibit the expression of OASL and IFIT2 may become a novel therapeutic approach for SLE.

Sequential activation of protein kinase C delta and JNK is required for interferon-alpha-induced expression of IFIT4.

A multitude of interferon (IFN)-inducible genes (IFIGs) are coordinately expressed in peripheral blood mononuclear cells (PBMCs) of patients with systemic lupus erythematosus (SLE), emphasising the globle activating of signal pathway mediated by IFN-I in SLE. In this study, we investigated the mechanisms of expression regulation of IFIT4 (interferon induced protein with tetratricopeptide repeats 4) by IFN-alpha. We found that IFN-alpha failed in inducing IFIT4 in STAT1-negative U3A cells. Ectopic expression of STAT1, but not mutant STAT1-S727A, almost completely restored IFN-alpha2a-induced IFIT4 expression. IFN-alpha induced the expression of IFIT4 and STAT1 in THP-1 cells, and this process was significantly antagonized by the specific inhibitors of both PKCdelta and JNK or their dominant negative mutants respectively. The inhibition of JNK activity by its specific inhibitor or its dominant negative mutant suppressed both IFIT4 expression and serine phosphorylation of STAT1 but not the activation of PKCdelta, while inhibition of PKCdelta suppressed activation of IFIT4, STAT1, and JNK. Our results suggest that the induction of IFIT4 transcription by IFN-alpha depends upon sequential activation of PKCdelta, JNK and STAT1, and that the influence of PKCdelta or JNK on IFN-alpha-mediated induction of IFIT4 is dependent upon the phosphorylation of STAT1 at Ser-727. The results in our experiment provide an in vitro model of the signaling mechanisms of IFIGs regulated by IFN-alpha, that is putatively thought to occur in vivo as the one of pathogenesis of SLE.