A Label-Free Optical Biosensor Based on an Array of Microring Resonators for the Detection of Human Serum Albumin.

We introduced a label-free sensing system based on an array of microring resonators (MRRs) which was successfully employed for human serum albumin (HSA) detection. The sensing-ring surface was functionalized to immobilize anti-HSA, facilitating HSA binding. Our refractive index sensing system demonstrates high sensitivity at 168 nm/RIU and a low limit of detection (LOD) of 63.54 ng/mL, closely comparable to current HSA detection methods. These findings confirm the potential of MRRs as biocompatible sensors for HSA detection. This system holds great promise as an innovative platform for the detection of HSA, carrying significant importance in medical diagnostics.

Targeted delivery of a PD-1-blocking scFv by CD133-specific CAR-T cells using nonviral Sleeping Beauty transposition shows enhanced antitumour efficacy for advanced hepatocellular carcinoma.

BACKGROUND: CD133 is considered a marker for cancer stem cells (CSCs) in several types of tumours, including hepatocellular carcinoma (HCC). Chimeric antigen receptor-specific T (CAR-T) cells targeting CD133-positive CSCs have emerged as a tool for the clinical treatment of HCC, but immunogenicity, the high cost of clinical-grade recombinant viral vectors and potential insertional mutagenesis limit their clinical application. METHODS: CD133-specific CAR-T cells secreting PD-1 blocking scFv (CD133 CAR-T and PD-1 s cells) were constructed using a sleeping beauty transposon system from minicircle technology, and the antitumour efficacy of CD133 CAR-T and PD-1 s cells was analysed in vitro and in vivo. RESULTS: A univariate analysis showed that CD133 expression in male patients at the late stage (II and III) was significantly associated with worse progression-free survival (PFS) (P = 0.0057) and overall survival (OS) (P = 0.015), and a multivariate analysis showed a trend toward worse OS (P = 0.041). Male patients with advanced HCC exhibited an approximately 20-fold higher PD-L1 combined positive score (CPS) compared with those with HCC at an early stage. We successfully generated CD133 CAR-T and PD-1 s cells that could secrete PD-1 blocking scFv based on a sleeping beauty system involving minicircle vectors. CD133 CAR-T and PD-1 s cells exhibited significant antitumour activity against HCC in vitro and in xenograft mouse models. Thus, CD133 CAR-T and PD-1 s cells may be a therapeutically tractable strategy for targeting CD133-positive CSCs in male patients with advanced HCC. CONCLUSIONS: Our study provides a nonviral strategy for constructing CAR-T cells that could also secrete checkpoint blockade inhibitors based on a Sleeping Beauty system from minicircle vectors and revealed a potential benefit of this strategy for male patients with advanced HCC and high CD133 expression (median immunohistochemistry score > 2.284).

A bioresponsive diselenide-functionalized hydrogel with cascade catalytic activities for enhanced local starvation- and hypoxia-activated melanoma therapy.

Glutathione (GSH) consumption-enhanced cancer therapies represent important potential cancer treatment strategies. Herein, we developed a new multifunctional diselenide-crosslinked hydrogel with glutathione peroxidase (GPx)-like catalytic activity for GSH depletion-enhanced glucose oxidase (GOx)-mediated tumor starvation and hypoxia-activated chemotherapy. By increasing acid and H2O2 during GOx-induced tumor starvation, the degradation of the multiresponsive scaffold could be promoted, which led to accelerated release of the loaded drugs. Meanwhile, the overproduced H2O2 led to accelerated intracellular GSH consumption under the cascade catalysis of small molecular selenides released from the degraded hydrogel, further enhancing the curative effect of in situ H2O2 and subsequent multimodal cancer treatment. Following the GOx-induced amplification of hypoxia, tirapazamine (TPZ) was transformed into the highly toxic benzotriazinyl radical (BTZ·), exhibiting enhanced antitumor activity. This GSH depletion-augmented cancer treatment strategy effectively boosted GOx-mediated tumor starvation and activated the hypoxia drug, leading to significantly enhanced local anticancer efficacy. STATEMENT OF SIGNIFICANCE: There has been a growing interest in depleting intracellular GSH as a potential strategy for improving ROS-based cancer therapy. Herein, a bioresponsive diselenide-functionalized dextran-based hydrogel with GPx-like catalytic activity was developed for GSH consumption-enhanced local starvation- and hypoxia-activated melanoma therapy. Results showed that the overproduced H2O2 led to accelerated intracellular GSH consumption under the cascade catalysis of small molecular selenides released from the degraded hydrogel, further enhancing the curative effect of in situ H2O2 and subsequent multimodal cancer treatment.

Cancer-Associated Fibroblast-Mediated Cellular Crosstalk Supports Hepatocellular Carcinoma Progression.

BACKGROUND AND AIMS: Cancer-associated fibroblasts (CAFs) are key players in multicellular, stromal-dependent alterations leading to HCC pathogenesis. However, the intricate crosstalk between CAFs and other components in the tumor microenvironment (TME) remains unclear. This study aimed to investigate the cellular crosstalk among CAFs, tumor cells, and tumor-associated neutrophils (TANs) during different stages of HCC pathogenesis. APPROACH AND RESULTS: In the HCC-TME, CAF-derived cardiotrophin-like cytokine factor 1 (CLCF1) increased chemokine (C-X-C motif) ligand 6 (CXCL6) and TGF-ß secretion in tumor cells, which subsequently promoted tumor cell stemness in an autocrine manner and TAN infiltration and polarization in a paracrine manner. Moreover, CXCL6 and TGF-ß secreted by HCC cells activated extracellular signal-regulated kinase (ERK) 1/2 signaling of CAFs to produce more CLCF1, thus forming a positive feedback loop to accelerate HCC progression. Inhibition of ERK1/2 or CLCF1/ciliary neurotrophic factor receptor signaling efficiently impaired CLCF1-mediated crosstalk among CAFs, tumor cells, and TANs both in vitro and in vivo. In clinical samples, up-regulation of the CLCF1-CXCL6/TGF-ß axis exhibited a marked correlation with increased cancer stem cells, "N2"-polarized TANs, tumor stage, and poor prognosis. CONCLUSIONS: This study reveals a cytokine-mediated cellular crosstalk and clinical network involving the CLCF1-CXCL6/TGF-ß axis, which regulates the positive feedback loop among CAFs, tumor stemness, and TANs, HCC progression, and patient prognosis. These results may support the CLCF1 cascade as a potential prognostic biomarker and suggest that selective blockade of CLCF1/ciliary neurotrophic factor receptor or ERK1/2 signaling could provide an effective therapeutic target for patients with HCC.

The efficacy and safety of the combination of axitinib and pembrolizumab-activated autologous DC-CIK cell immunotherapy for patients with advanced renal cell carcinoma: a phase 2 study.

OBJECTIVES: Although axitinib has achieved a preferable response rate for advanced renal cell carcinoma (RCC), patient survival remains unsatisfactory. In this study, we evaluated the efficacy and safety of a combination treatment of axitinib and a low dose of pembrolizumab-activated autologous dendritic cells-co-cultured cytokine-induced killer cells in patients with advanced RCC. METHODS: All adult patients, including treatment-naive or pretreated with VEGF-targeted agents, were enrolled from May 2016 to March 2019. Patients received axitinib 5 mg twice daily and pembrolizumab-activated dendritic cells-co-cultured cytokine-induced killer cells intravenously weekly for the first four cycles, every 2 weeks for the next four cycles, and every month thereafter. RESULTS: The 43 patients (22 untreated and 21 previously treated) showed a median progression-free survival (mPFS) of 14.7 months (95% CI, 11.16-18.30). mPFS in treatment-naive patients was 18.2 months, as compared with 14.4 months in pretreated patients (log-rank P-value = 0.07). Overall response rates were 25.6% (95% CI, 13.5-41.2%). Grade 3 or higher adverse events occurred in 5% of patients included hypertension (11.6%) and palmar-plantar erythrodysesthesia (7.0%). Peripheral blood lymphocyte immunophenotype and serum cytokine profile analyses demonstrated increased antitumor immunity after combination treatment particularly in patients with a long-term survival benefit, while those with a minimal survival benefit demonstrated an elevated proportion of peripheral CD8+TIM3+ T cells and lower serum-level immunostimulatory cytokine profile. CONCLUSIONS: The combination therapy was active and well tolerated for treatment of advanced RCC, either as first- or second-line treatment following other targeted agents. Changes in immunophenotype and serum cytokine profile may be used as prognostic biomarkers.

Development and validation of a clinical score to predict late seizures after intracerebral hemorrhage in Chinese.

BACKGROUND: Seizures are a frequent complication after intracerebral hemorrhage (ICH). The CAVE score was developed in Europeans to predict late seizures after ICH. Given the higher incidence of ICH in Asians, we aimed to develop and validate a clinical scoring tool for predicting post-ICH late seizures in Chinese. METHODS: We retrospectively included patients admitted with ICH to a major stroke center in Shandong province, China, in the derivation cohort, who were followed up for occurrence of late seizures (more than seven days after ICH). We applied Cox regression model to identify significant clinical factors which were used to derive a predictive scoring model. The performance of this model was compared with CAVE, and validated in a separate cohort of patients with ICH admitted to another stroke center. RESULTS: In the derivation cohort (n = 602; median age 65 years; 57 % male；median follow up 24 months), 47 (7.8 %) patients had late seizures during follow up. Four significant risk factors were identified and selected to derive the LANE score (Lobar hemorrhage, Age <65 years, NIHSS score ≥15, Early seizures). The total possible points ranged from 0 to 6, corresponding to positive predictive values of 10.1%-100%, and negative predictive values of 96.8%-92.2%, respectively. The c-statistics of the LANE score in the derivation cohort and validation cohort (n = 521) were 0.83 and 0.78, respectively, while those of the CAVE score were 0.81 and 0.74, respectively. CONCLUSION: We have developed and validated a clinical scoring tool for predicting late seizures after ICH in Chinese. This tool may be used to identify high risk patients for closer monitoring and clinical trials of therapies to prevent post-ICH epilepsy in the future.

Integration of PET/CT Radiomics and Semantic Features for Differentiation between Active Pulmonary Tuberculosis and Lung Cancer.

PURPOSE: We aim to accurately differentiate between active pulmonary tuberculosis (TB) and lung cancer (LC) based on radiomics and semantic features as extracted from pre-treatment positron emission tomography/X-ray computed tomography (PET/CT) images. PROCEDURES: A total of 174 patients (77/97 pulmonary TB/LC as confirmed by pathology) were retrospectively selected, with 122 in the training cohort and 52 in the validation cohort. Four hundred eighty-seven radiomics features were initially extracted to quantify phenotypic characteristics of the lesion region in both PET and CT images. Eleven semantic features were additionally defined by two experienced nuclear medicine physicians. Feature selection was performed in 5 steps to enable derivation of robust and effective signatures. Multivariable logistic regression analysis was subsequently used to develop a radiomics nomogram. The calibration, discrimination, and clinical usefulness of the nomogram were evaluated in both the training and independent validation cohorts. RESULTS: The individualized radiomics nomogram, which combined PET/CT radiomics signature with semantic features, demonstrated good calibration and significantly improved the diagnostic performance with respect to the semantic model alone or PET/CT signature alone in training cohort (AUC 0.97 vs. 0.94 or 0.91, p = 0.0392 or 0.0056), whereas did not significantly improve the performance in validation cohort (AUC 0.93 vs. 0.89 or 0.91, p = 0.3098 or 0.3323). CONCLUSION: The radiomics nomogram showed potential for individualized differential diagnosis between solid active pulmonary TB and solid LC, although the improvement of performance was not significant relative to semantic model.

Hypoxia Induces Mitochondrial Defect That Promotes T Cell Exhaustion in Tumor Microenvironment Through MYC-Regulated Pathways.

T cell exhaustion is an obstacle to immunotherapy for solid tumors. An understanding of the mechanism by which T cells develop this phenotype in solid tumors is needed. Here, hypoxia, a feature of the tumor microenvironment, causes T cell exhaustion (TExh) by inducing a mitochondrial defect. Upon exposure to hypoxia, activated T cells with a TExh phenotype are characterized by mitochondrial fragmentation, decreased ATP production, and decreased mitochondrial oxidative phosphorylation activity. The TExh phenotype is correlated with the downregulation of the mitochondrial fusion protein mitofusin 1 (MFN1) and upregulation of miR-24. Overexpression of miR-24 alters the transcription of many metabolism-related genes including its target genes MYC and fibroblast growth factor 11 (FGF11). Downregulation of MYC and FGF11 induces TExh differentiation, reduced ATP production and a loss of the mitochondrial mass in T cell receptor (TCR)-stimulated T cells. In addition, we determined that MYC regulates the transcription of FGF11 and MFN1. In nasopharyngeal carcinoma (NPC) tissues, the T cells exhibit an increased frequency of exhaustion and loss of mitochondrial mass. In addition, inhibition of miR-24 signaling decreases NPC xenograft growth in nude mice. Our findings reveal a mechanism for T cell exhaustion in the tumor environment and provide potential strategies that target mitochondrial metabolism for cancer immunotherapy.

Efficacy of adjuvant cytokine-induced killer cell immunotherapy in patients with colorectal cancer after radical resection.

Adjuvant chemotherapy after surgery is the standard treatment modality for stage III and part of stage II or stage IV colorectal cancer (CRC) patients. However, the 5-year overall survival (OS) rate remains unsatisfactory. Thus, developing combination therapies is essential to improve the prognosis of patients with CRC. The present study aimed to determine the effect of a sequential combination of cytokine-induced killer cell (CIK) infusion and chemotherapy for patients with CRC. 122 patients with CRC treated with postoperative adjuvant chemotherapy were retrospectively included in this study. Among them, 62 patients received adjuvant chemotherapy only (control group), while the other 60 patients, with similar demographic and clinical characteristics, received adjuvant chemotherapy and sequential CIK cell immunotherapy (CIK group). Survival analysis showed significantly improved disease free survival (DFS) and OS rates in the CIK group compared with the control group (log-rank test, P = .0024; P = .008, respectively). Univariate and multivariate analyses indicated that sequential CIK cell treatment was an independent prognostic factor for patients' DFS and OS. Subgroup analyses showed that sequential CIK cell treatment significantly improved the DFS and OS of patients with high-risk T4 stage and insufficient chemotherapy duration. In conclusion, these data indicate that sequential adjuvant CIK cell treatment combined with chemotherapy is an effective therapeutic strategy to prevent disease recurrence and prolong survival of patients with CRC, particularly for patients with high-risk T4 stage and insufficient chemotherapy duration.

[18F-FDG PET/CT manifestations of massive type active pulmonary tuberculosis and its differentiation from lung cancer].

OBJECTIVE: To investigate 18F-FDG PET/CT manifestations of massive type active tuberculosis and lung cancer and the differential diagnosis of the two diseases based on 18F-FDG PET/CT findings. METHODS: We retrospectively collected the data from 74 patients with active tuberculosis and 64 patients with lung cancer, whose lesions presented as solid masses on CT. The demographic and clinical data of the patients, 18F-FDG PET characteristics including SUVmax, 18F-FDG uptake (higher than mediastinal blood pool or not), radioactive defect within the lesion, and the CT findings including the lesion size, signs of cavity, vacuoles, lobulation, smooth border, and mediastinal/lung window ratio (M/L ratio) of the lesions were analyzed. Univariate and multivariate analyses were used to compare the variables between the two groups, and a logistic regression model was established for differentiation of the two diseases. The diagnostic efficiency was evaluated by area under the receiver-operating characteristic (ROC) curve analysis. RESULTS: No significant differences were found in the quantitative index (SUVmax >2.5 or not) or in the qualitative index (uptake of lesion higher than mediastinal blood pool or not) in PET between massive type active tuberculosis and lung cancer (P>0.05). Univariate analysis revealed that SUVmax, 18F-FDG uptake of the lesion, age, lesion size, signs of cavity, or M/L ratio were not significantly different (P>0.05), but gender, signs of radioactive defect, vacuoles, smooth border and lobulation were significantly different (P < 0.05) between the two diseases. Multivariate analysis showed that gender, signs of radioactive defect, smooth border and lobulation of the lesion were independent factors for discrimination of the two diseases (P < 0.05). A risk prediction model for active tuberculosis was established based on logistic regression analysis: P=1/(1+e-x), X=-0.530+1.978×gender+3.343×radioactive defect +2.846×smooth border-2.116×lobulation. For diagnosis of active tuberculosis, the sensitivity, specificity, accuracy, positive predictive value and negative predictive value of this model were 78.4%, 92.2%, 84.8%, 92.1%, and 78.7%, respectively. CONCLUSIONS: The combined analysis of gender, signs of radioactive defect, smooth border and lobulation of the lesions is useful for discriminating massive type active tuberculosis from lung cancer in the majority of the patients, whereas 18F-FDG uptake alone has only limited value for a differential diagnosis.

Benefits of 18F-FDG PET/CT for the preoperative characterisation or staging of disease in the ampullary and duodenal papillary.

OBJECTIVES: Diagnosing ampullary and duodenal papillary carcinomas (ADPCs) is challenging. In the present study, we investigated the application value of 18F-fluorodeoxyglucose positron emission tomography/computed tomography (18F-FDG PET/CT) in the preoperative evaluation of these tumours. METHODS: 18F-FDG PET/CT images of 58 patients with ADPC and 28 patients with benign disease were retrospectively analysed. Preoperative 18F-FDG PET/CT was compared to contrast-enhanced (CE) CT and magnetic resonance imaging (MRI) in terms of diagnostic efficacy, certainty, staging and impact on treatment decisions. RESULTS: 18F-FDG PET/CT showed a high sensitivity (93.1%) and a medium specificity (78.6%) for diagnosing ADPC. Compared to CE CT/MRI, 18F-FDG PET/CT had a higher diagnostic specificity (78.6 vs. 35.7%, p = 0.001) but a similar sensitivity (93.1 vs. 89.6%, p = 0.508). 18F-FDG PET/CT provided a much higher diagnostic certainty than CE CT/MRI (definite reports, 88.4 vs. 50.0%, χ2 = 29.698, p < 0.001), especially for small tumours ≤ 1.5 cm, and found distant metastases in five patients. The 18F-FDG PET/CT findings affected the treatment plans of 11 patients and improved the confidence in the diagnoses of 28 patients. CONCLUSIONS: The present study demonstrated that 18F-FDG PET/CT can supplement CE CT/MRI to provide a more accurate diagnosis for ADPC, and thus, plays an important role in the decision-making process before complicated pancreaticoduodenectomy procedures. KEY POINTS: • It is a challenge for CT and MRI to diagnose ampullary carcinoma, especially at their early stage. • Our study demonstrated that the benefit of PET/CT was improving the diagnostic confidence for ampullary and duodenal papillary carcinomas. • 18F-FDG PET/CT can change the treatment decision for ampullary and duodenal papillary carcinomas.

Retrospective analysis of the efficacy of cytokine-induced killer cell immunotherapy combined with first-line chemotherapy in patients with metastatic colorectal cancer.

OBJECTIVES: Fluoropyrimidine-based chemotherapy regimens are the current first-line treatment for metastatic colorectal cancer (mCRC); however, the outcome is often unsatisfactory. The present study aimed to determine the effect of combined cytokine-induced killer (CIK) cell immunotherapy and first-line chemotherapy in patients with mCRC. METHODS: This retrospective study included 252 patients with mCRC treated with first-line chemotherapy. Among them, 126 patients received first-line chemotherapy only (control group), while the other 126 patients, with similar demographic and clinical characteristics, received CIK cell immunotherapy combined with first-line chemotherapy (CIK group). Overall survival (OS) and progression-free survival (PFS) were compared between the two groups using the Kaplan-Meier method. RESULTS: The median OS for the CIK group was 54.7 versus 24.1 months for the controls, and the median PFS for the CIK group was 25.7 versus 14.6 months for the controls. Univariate and multivariate analyses indicated that CIK cell treatment was an independent prognostic factor for patients' OS and PFS. Subgroup analyses showed that CIK cell treatment significantly improved the OS and PFS of patients with metastatic colon cancer, but not those with metastatic rectal cancer. Additionally, the change in CD3+CD56+ subsets after the fourth treatment cycle might be an indicator of successful CIK cell treatment: Patients with increased CD3+CD56+ subsets had better survival than those with decreased CD3+CD56+ subsets. CONCLUSION: Cytokine-induced killer cell immunotherapy combined with first-line chemotherapy could significantly improve the OS and PFS of patients with mCRC, particularly for patients with metastatic colon cancer.

CIK cell cytotoxicity is a predictive biomarker for CIK cell immunotherapy in postoperative patients with hepatocellular carcinoma.

Adjuvant cytokine-induced killer (CIK) cell immunotherapy has shown potential in improving the prognosis of hepatocellular carcinoma (HCC) patients after curative resection. However, whether an individual could obtain survival benefit from CIK cell treatment remains unknown. In the present study, we focused on the characteristics of CIK cells and aimed to identify the best predictive biomarker for adjuvant CIK cell treatment in patients with HCC after surgery. This study included 48 patients with HCC treated with postoperative adjuvant CIK cell immunotherapy. The phenotype activity and cytotoxic activity of CIK cells were determined by flow cytometry and xCELLigence™ Real-Time Cell Analysis (RTCA) system, respectively. Correlation analysis revealed that the cytotoxic activity of CIK cells was significantly negative correlated with the percentage of CD3+ CD4+ cell subsets, but significantly positive correlated with CD3-CD56+ and CD3+ CD56+ cell subsets. Survival analysis showed that there were no significant associations between patients' prognosis and the phenotype of CIK cells. By contrast, there was statistically significant improvement in recurrence-free survival (RFS) and overall survival (OS) for patients with high cytotoxic activity of CIK cells as compared with those with low cytotoxic activity of CIK cells. Univariate and multivariate analyses indicated that CIK cell cytotoxicity was an independent prognostic factor for RFS and OS. In conclusion, a high cytotoxic activity of CIK cells can serve as a valuable biomarker for adjuvant CIK cell immunotherapy of HCC patients after surgery.

18F-FDG PET/CT imaging findings in anaplastic large cell lymphoma, a rare subtype of lymphoma.

OBJECTIVE: To investigate the 18F-FDG PET/CT imaging manifestations for anaplastic large cell lymphoma (ALCL), a rare subtype of T/NK cell lymphoma. METHODS: Fifty patients with ALCL, including 32 anaplastic lymphoma kinase (ALK)-positive patients and 18 ALK-negative patients, were enrolled. The positive detection, maximal standardized uptake value (SUVmax), and distribution of nodal and extranodal involvement were recorded and analysed. Fifty patients with diffuse large B cell lymphoma (DLBCL) were collected as a control group. RESULTS: ALCL lesions were demonstrated to be 18F-FDG-avid tumours with a mean SUVmax of 19.4 ± 12.6. Most (76%) ALCL patients presented with stage III-IV disease, and nodal and extranodal involvement occurred in 74.0 and 72.0% of the patients, respectively. ALCL and DLBCL showed many similarities in tumour stage, 18F-FDG uptake and tumour involvement (P > 0.05), although the preferred extranodal organs of involvement (bone and the gastrointestinal tract, respectively) were different (P < 0.05). Compared to ALK-negative lesions, a higher uptake of 18F-FDG was found in the ALK-positive lesions (SUVmax: 22.1 ± 14.3 vs. 15.1 ± 6.6, t = 2.354, P = 0.023). ALK-positive ALCL was more likely to involve the lymph nodes than ALK-negative ALCL (84.3% vs. 55.5%, χ2 = 4.973, P = 0.043), while ALK-negative ALCL was more prone to involve the extranodal organs compared to ALK-positive ALCL (88.9% vs. 62.5%, χ2 = 3.979, P = 0.046). CONCLUSION: The present study demonstrated that ALCL is a systemic 18F-FDG-avid lymphoma with many imaging manifestations similar to DLBCL on PET/CT. The present study also showed that ALK expression actually influenced tumour 18F-FDG uptake and lesion distribution. These findings may be useful to improve the understanding of the biological characteristics of ALCL.

Radiofluorinated GPC3-Binding Peptides for PET Imaging of Hepatocellular Carcinoma.

PURPOSE: Hepatocellular carcinoma (HCC) remains one of the most challenging diseases worldwide. Glypican-3 (GPC-3) is a cell surface proteoglycan that is overexpressed on the membrane of HCC cells. The purpose of this study was to develop a target-specific radiofluorinated peptide for positron emission tomography (PET) imaging of GPC3 expression in hepatocellular carcinoma. PROCEDURES: New GPC3-binding peptides (GP2076 and GP2633) were radiolabeled with F-18 using Al[18F]F labeling approach, and the resulting PET probes were subsequently subject to biological evaluations. A highly hydrophilic linker was incorporated into GP2633 with an aim of reducing the probe uptake in liver and increasing tumor-to-liver (T/L) contrast. Both GP2076 and GP2633 were radiolabeled using Al[18F]F chelation approach. The binding affinity, octanol/water partition coefficient, cellular uptake and efflux, and stability of both F-18 labeled peptides were tested. Tumor targeting efficacy and biodistribution of Al[18F]F-GP2076 and Al[18F]F-GP2633 were determined by PET imaging in HCC-bearing mice. Immunohistochemistry analyses were performed to compare the findings from PET scans. RESULTS: Al[18F]F-GP2076 and Al[18F]F-GP2633 were rapidly radiosynthesized within 20 min in excellent radiochemical purity (> 97 %). Al[18F]F-GP2633 was determined to be more hydrophilic than Al[18F]F-GP2076 in terms of octanol/water partition coefficient. Both Al[18F]F-GP2076 and Al[18F]F-GP2633 demonstrated good in vitro and in vivo stability and binding specificity to GPC3-positive HepG2 cells. For PET imaging, Al[18F]F-GP2633 exhibited enhanced uptake in HepG2 tumor (%ID/g 3.37 ± 0.35 vs. 2.13 ± 0.55, P = 0.031) and reduced accumulation in liver (%ID/g 1.70 ± 0.26 vs. 3.70 ± 0.98, P = 0.027) at 60 min post-injection (pi) as compared to Al[18F]F-GP2076, resulting in significantly improved tumor-to-liver (T/L) contrast (ratio 2.00 ± 0.18 vs. 0.59 ± 0.14, P = 0.0004). Higher uptake of Al[18F]F-GP2633 in GPC3-positive HepG2 tumor was observed as compared to GPC3-negative McA-RH7777 tumor (%ID/g 3.37 ± 0.35 vs. 1.64 ± 0.03, P = 0.001) at 60 min pi, confirming GPC3-specific accumulation of Al[18F]F-GP2633 in HepG2 tumor. CONCLUSION: The results demonstrated that Al[18F]F-GP2633 is a promising probe for PET imaging of GPC3 expression in HCC. Convenient preparation, excellent GPC3 specificity in HCC, and favorable excretion profile of Al[18F]F-GP2633 warrant further investigation for clinical translation. PET imaging with a GPC3-specific probe would provide clinicians with vital diagnostic information that could have a significant impact on the management of HCC patients.

A green separation mode of synephrine from Citrus aurantium L. (Rutaceae) by nanofiltration technology.

Thermal breakage of alkaloid ingredients was a common problem to which attention should be paid in the application of fruit ingredients separation. In this study, the mathematical models were established to predict the rejection of synephrine from Citrus aurantium L. (Rutaceae). The experiment showed that there was a linear relationship between operation pressure and membrane flux. Meanwhile, under the influence of solution-diffusion effect and the charge effect, the mass transfer coefficient was power functioned with initial concentration. The mathematical model showed that the predicted rejections of synephrine from Citrus aurantium extract were well approximate to real ones, and the lipid-lowering active ingredient had effectively enriched. The predicted model of nanofiltration separation has a preferable applicability to synephrine and provides references for nanofiltration separation, especially for raw food materials with synephrine.

[Correlation of molecular weight and nanofiltration mass transfer coefficient of phenolic acid composition from Salvia miltiorrhiza].

Based on the molecular sieving and solution-diffusion effect in nanofiltration separation, the correlation between initial concentration and mass transfer coefficient of three typical phenolic acids from Salvia miltiorrhiza was fitted to analyze the relationship among mass transfer coefficient, molecular weight and concentration. The experiment showed a linear relationship between operation pressure and membrane flux. Meanwhile, the membrane flux was gradually decayed with the increase of solute concentration. On the basis of the molecular sieving and solution-diffusion effect, the mass transfer coefficient and initial concentration of three phenolic acids showed a power function relationship, and the regression coefficients were all greater than 0.9. The mass transfer coefficient and molecular weight of three phenolic acids were negatively correlated with each other, and the order from high to low is protocatechualdehyde >rosmarinic acid> salvianolic acid B. The separation mechanism of nanofiltration for phenolic acids was further clarified through the analysis of the correlation of molecular weight and nanofiltration mass transfer coefficient. The findings provide references for nanofiltration separation, especially for traditional Chinese medicine with phenolic acids.

A UPLC-MS/MS method for simultaneous determination of 1-deoxynojirimycin and N-methyl-1-deoxynojirimycin in rat plasma and its application in pharmacokinetic and absolute bioavailability studies.

A specific, sensitive, rapid, precise, and reliable UPLC-MS/MS-based method was designed for the first time for the simultaneous determination of 1-deoxynojirimycin (DNJ) and N-methyl-1-deoxynojirimycin (N-CH3-DNJ) in rat plasma. Miglitol was served as the internal standard (IS). An MN-NUCLEODUR HILIC column was assessed to separate the two compounds by isocratic elution using acetonitrile: water with 0.05% formic acid and 6.5mM ammonium acetate (72:28, v/v) at a flow rate of 0.4mL/min. A triple quadrupole mass spectrometer was operated in the positive ionization mode using multiple reaction monitoring (MRM), and it was employed to determine transitions of m/z 164.1→110.1, 178.1→100.1, and 208.1→146.1 for DNJ, N-CH3-DNJ, and IS, respectively. The method of the two constituents was validated and the results were acceptable. The absolute bioavailability of DNJ and N-CH3-DNJ in rats was 50±9% and 62±24%, respectively. The method was then successfully used for the first time to study the pharmacokinetic behavior and absolute bioavailability of DNJ and N-CH3-DNJ in rats after intravenous (10mg/kg) and oral administration (80mg/kg). The results of this study might provide more information on preclinical pharmacokinetics and a solid basis for assessing the clinical efficacy of DNJ and N-CH3-DNJ.

[Optimize concentrate process of alkaloid from Leonurus japonicus by ultrafultration-nanofiltration coupling technology].

To optimize the concentrate process of alkaloid from Leonurus japonicus by nanofiltration-ultrafiltration coupling technology with response surface methodology. The experiment showed that after ultrafiltration pre-treatment, the total protein removal rate was 94.38% in aqueous extract from L. japonicus, and the nanofiltration technology had obvious advantages over the conventional concentrate process. The optimal concentrate conditions were as follows：molecular weight cut-off 450, pH 3.07, concentration of stachydrine hydrochloride 80.15 mg•L⁻¹, and concentration of the total alkaloid 285.73 mg•L⁻¹. The cut-off rate was 93.37% and 95.85% respectively for stachydrine hydrochloride and the total alkaloid under the optimum conditions, with a relative error of 0.79% and 1.16% respectively. The combination of Box-Behnken design and response surface analysis can well optimize the concentrate process of L. japonicus by nanofiltration, and the results provide the basis for nanofiltration concentrate for heat-sensitive traditional Chinese medicine.