Solanine induces ferroptosis in colorectal cancer cells through ALOX12B/ADCY4 molecular axis.

OBJECTIVES: Colorectal cancer (CRC) is the fourth most commonly diagnosed cancer worldwide. Solanine is a phytochemical extracted from traditional Chinese medicine with widely reported anticancer effects. Here, we investigated the potential role of solanine in regulating ferroptosis in CRC cells and scrutinized the molecular mechanism. METHODS: Cell growth and cytotoxicity were examined using CCK-8 proliferation assay and lactate dehydrogenase assay. Oxidative stress was determined by measuring glutathione (GSH), malondialdehyde, and reactive oxygen species (ROS) levels. Subcellular changes in mitochondria were examined by transmission electron microscopy. Gene and protein expression levels were detected by quantitative real-time polymerase chain reaction (qRT-PCR). Protein-protein interaction was determined by co-immunoprecipitation. KEY FINDINGS: Solanine arrested cell proliferation in CRC cells and induced typical ferroptotic changes. Solanine treatment promoted ROS production, lipid peroxidation, and cell membrane disruption, while the cellular level of antioxidant GSH was reduced upon solanine treatment. ALOX12B was identified as a molecular mediator of solanine to promote ferroptosis. Solanine treatment upregulated ALOX12B levels and silencing ALOX12B could suppress solanine-induced ferroptosis. Further, ADCY4 was found to physically associate with ALOX12B and maintain ALOX12B protein stability. Silencing ADCY4 destabilized ALOX12B and attenuated solanine-induced ferroptosis. CONCLUSIONS: Our data demonstrated the ferroptosis-inducing effect of solanine in CRC cells, and revealed ALOX12B/ADCY4 molecular axis as the ferroptosis mediator of solanine. Solanine may synergize with existing ferroptosis inducer as an anticancer strategy in CRC, which warrants further validation in animal experiments.

Survival Contradiction in Stage II, IIIA, And IIIB Colon Cancer: A Surveillance, Epidemiology, and End Result-Based Analysis.

There exists an inconsistency between stage and survival in the current American Joint Committee on Cancer (AJCC) staging system for colon cancer. In this study, we compared the clinicopathological characteristics and prognosis of colon cancer patients with stage II, IIIA, and IIIB disease based on the surveillance, epidemiology, and end results (SEER) database. Kaplan-Meier analysis was used to generate overall survival (OS) and cancer-specific survival (CSS) curves. The Cox regression was employed to identify risk factors. The competing risk model was completed by the cumulative incidence function and Gray's test. In the final population of 31,361 colon cancer patients, Kaplan-Meier curve analysis showed that stage IIIA had the highest OS and CSS, followed by stage IIA and IIIB, and IIB and IIC showed the worst OS and CSS. In the Cox model, the stage was proven to be an independent prognostic factor. In the competing risk model, stage IIIA colon cancer patients had the lowest 5-year cancer-specific death rate in stages II, IIIA, and IIIB. In conclusion, the prognosis of colon cancer patients in stage IIA was worse than that of patients in stage IIIA, while the survival rate of stage IIB and IIC was lower than that of stage IIIB.

[Preparation of molecularly imprinted polymers-functionalized silica nanoparticles for the separation and recognition of aristolochic acids].

Aristolochic acids (AAs), which is commonly found in Aristolochia and Asarum plants, has been widely used in several traditional medicine practices due to their anti-inflammatory, anti-malarial, and anti-hyperglycemic activities. Recently, researchers have found a “decisive link” between liver cancer and aristolochic acid after analyzing a large number of liver cancer samples around the world. Therefore, a highly sensitive and selective method is required for the analysis of AAs in traditional Chinese medicines (TCM). For the determination of AAs in TCM, pretreatment is indispensable because in actual TCM samples, AAs is present in trace amounts and the complex matrix exerts interference. In the past decades, molecularly imprinted polymers (MIPs) have attracted considerable attention as an alternative for the trace analysis in complicated matrices. In this study, MIP-coated SiO2 nanoparticles (SiO2@MIP NPs) was prepared for the determination of aristolochic acid by surface molecular imprinting using aristolochic acid Ⅰ (AAI ) as the template molecule, 2-vinylpyridine (VPY) as the functional monomer, and ethyleneglycol dimethacrylate (EGDMA) as the cross-linking agent. Core-shell-structure SiO2@MIP NPs were obtained by modifying vinyl groups on the surface of SiO2 NPs, coating MIPs films onto the silica surface via selective polymerization, and final extraction of template AAI and generation of the recognition site. To find a suitable functional monomer for the best imprinting effect, the interaction between the template and the functional monomers, including acrylic acid (AA), methyl acrylic acid (MAA), 2-vinyl pyridine (VPY), acrylamide (AM), and methylacrylamide (MAM) was investigated. Electrostatic interaction between AAI and VPY resulted in the maximum decrease in absorbance of AAI at 250 nm. Therefore, VPY was chosen for the preparation of MIP. The morphological and physical properties of the MIPs were characterized by transmission electron microscopy (TEM), Fourier-transform infrared (FT-IR) spectroscopy, thermogravimetric analysis, and N2 adsorption and desorption surface analysis. TEM images showed that SiO2 NPs were monodispersed with diameter of about 200 nm. The clear core-shell structure of SiO2@MIP NPs was observed, and the thickness of MIPs coating was about 35 nm. The FT-IR spectra of SiO2 NPs, vinyl group modified SiO2 and SiO2@MIP NPs revealed that the vinyl group and organic MIP layer were successfully modified at SiO2 sequentially. The results of thermogravimetric analysis were consistent with the FT-IR data for different SiO2 NPs. The nitrogen gas adsorption-desorption experiments showed that SiO2@MIP NPs and non-imprinted polymer (SiO2@NIP NPs) have the same pore volumes, while the surface area and pore size of MIPs were slightly larger than those of NIPs. Therefore, the difference in adsorption between SiO2@MIP NPs and SiO2@NIP NPs resulted from the imprinted sites on the MIP surface, rather than the difference in their surface areas. The adsorption properties of SiO2@MIP NPs were demonstrated by kinetic, isothermal, and selective adsorption experiments. The results of these experiments displayed that SiO2@MIP NPs reached adsorption equilibrium within a short period (120 s) and possessed a much higher rebinding ability than SiO2@NIP NPs. To verify the selectivity of SiO2@MIP NPs for AAI, three structural analogues (viz. tanshinone ⅡA, 2-methoxy-5-nitrophenol, and benzoic acid) were selected. The results showed that the binding capacity of SiO2@MIP NPs was much higher than those of these analogues. SiO2@MIP NPs have high adsorption capacity (5.74 mg/g), high imprinting factor (4.9), good selectivity coefficient (2.3-6.6) towards the structural analogues. SiO2@MIP NPs was used as an adsorbent and combined with HPLC for the selective separation of AAI in TCM. The recoveries of Kebia trifoliate samples spiked with three levels of AAI (0.3, 0.5, and 1.0 μg/mL) ranged from 73% to 83%. The results suggested that the proposed SiO2@MIP NPs could be used for selective enrichment of AAI from real complex TCM samples.

A nomogram based on clinicopathological features and serological indicators predicting breast pathologic complete response of neoadjuvant chemotherapy in breast cancer.

A single tumor marker is not enough to predict the breast pathologic complete response (bpCR) after neoadjuvant chemotherapy (NAC) in breast cancer patients. We aimed to establish a nomogram based on multiple clinicopathological features and routine serological indicators to predict bpCR after NAC in breast cancer patients. Data on clinical factors and laboratory indices of 130 breast cancer patients who underwent NAC and surgery in First Affiliated Hospital of Xi'an Jiaotong University from July 2017 to July 2019 were collected. Multivariable logistic regression analysis identified 11 independent indicators: body mass index, carbohydrate antigen 125, total protein, blood urea nitrogen, cystatin C, serum potassium, serum phosphorus, platelet distribution width, activated partial thromboplastin time, thrombin time, and hepatitis B surface antibodies. The nomogram was established based on these indicators. The 1000 bootstrap resampling internal verification calibration curve and the GiViTI calibration belt showed that the model was well calibrated. The Brier score of 0.095 indicated that the nomogram had a high accuracy. The area under the curve (AUC) of receiver operating characteristic (ROC) curve was 0.941 (95% confidence interval: 0.900-0.982) showed good discrimination of the model. In conclusion, this nomogram showed high accuracy and specificity and did not increase the economic burden of patients, thereby having a high clinical application value.

A strategy of utilizing Cu2+-mediating interaction to prepare magnetic imprinted polymers for the selective detection of celastrol in traditional Chinese medicines.

In this work, a novel strategy to prepare molecularly imprinted polymers (MIPs) functionalized magnetic carbon nanotubes (MCNTs) via a facile sol-gel polymerization by adopting Cu2+-mediating interaction was presented for selective recognition of celastrol (Cel), in the traditional Chinese medicines (TCM). Firstly, template Cel, 3-aminopropyltriethoxysilane (APTES) as monomer and Cu2+ (co-monomer) were mixed to form a self-assembled pre-complex, in which Cu2+ could coordinate with Cel. Meanwhile, APTES plays a role of bridge between APTES and Cel. Secondly, carboxyl modified MCNTs as substrate was added into the pre-complex solution. After that, a multi-step sol-gel polymerization process was occurred in the presence of tetraethylorthosilicate as cross-linker and acetic acid as catalyst. Finally, MIPs layer was formed on the surface of the MCNTs (Cel-MIPs@MCNTs) after the removal of template with methanol/acetic. The morphology and structure of Cel-MIPs@MCNTs was investigated by various characterization techniques. The adsorption performance of Cel-MIPs@MCNTs to Cel was illustrated by kinetic, isothermal and selective binding experiments. The results displayed that the Cel-MIPs@MCNTs possessed fast kinetic equilibrium time (40 s), high adsorption capacity (13.35 µg mg-1), good imprinting factor of 3.41, and high magnetic responsivity (44.38 emu·g-1), which can be used as an ideal adsorbent for rapid isolation and enrichment of target analytes. A selective and sensitive method based on Cel-MIPs@MCNTs coupling with HPLC was developed for Cel determination including a wide linear range (0.15-200 µg mL-1) with correlation coefficient of 0.9998, a low limit of detection (0.05 µg mL-1). Furthermore, the applicability of Cel-MIPs@MCNTs was demonstrated to isolate and determine Cel in TCM samples with satisfactory recoveries ranged from 84.47% to 91.5% (RSD<5.35%). The results revealed that Cel-MIPs@MCNTs offer great potential as an adsorbent for selective and efficient isolation of Cel from complex TCM samples.

Hydrophilic molecularly imprinted polymers functionalized magnetic carbon nanotubes for selective extraction of cyclic adenosine monophosphate from winter jujube.

In this work, a green strategy was developed to prepare molecularly imprinted polymers functionalized magnetic carbon nanotubes in aqueous phase under mild conditions for cyclic adenosine monophosphate. Thanks to water solubility of chitosan, a natural polysaccharide which is rich in amino and hydroxyl groups, provided the feasibility to synthesize the green molecularly imprinted polymers for water soluble template in aqueous media. Coupled with high-performance liquid chromatography, the method exhibited a short equilibrium time (6 min), high adsorption capacity (22.42 µg/mg), high magnetic susceptibility, and good selectivity to template molecule with the imprinting factor of 2.94. A good linearity in the range of 0.020-3.0 mg/mL for target was obtained with a correlation coefficient of 0.9998. The limit of detection (signal-to-noise ratio = 3) and limit of quantitation (signal-to-noise ratio = 10) of the magnetic solid phase extraction method for cyclic adenosine monophosphate were 5 and 15 ng/mg, respectively. And the practical application of chitosan-based molecularly imprinted polymers as adsorbent to isolate and determine cyclic adenosine monophosphate in real natural samples (winter jujube) was demonstrated.

A dry powder inhalable formulation of salvianolic acids for the treatment of pulmonary fibrosis: safety, lung deposition, and pharmacokinetic study.

Salvianolic acids (SAL), the main bioactive component extracted from Salvia miltiorrhiza, is a natural product with a reported anti-pulmonary fibrosis (PF) effect. SAL is commonly administrated orally; however, it has a low oral bioavailability (less than 5%). The objective of this work was to develop a new dry powder inhalable formulation intended to facilitate the access of SAL to the target place. We prepared the new SAL powder formulation containing L-arginine and 2% of lecithin using the ball milling technique. L-arginine was used to regulate the strong acidity of the SAL solution, and lecithin was added to disperse the powder and improve the flowability. The resulting powder had a content in salvianolic acid B (SALB, the main active principle of SAL) of 66.67%, a particle size of less than 5 µm and a good flowability. In vivo fluorescence imaging showed that the powder could be successfully aerosolized and delivered to the lung. The acute lung irritation study proved that the presence of L-arginine improved the biocompatibility of the powder. Finally, according to the pharmacokinetic study, the new SAL powder formulation was found to significantly increase drug concentration in the lung and the bioavailability. In conclusion, the new dry powder inhalable formulation of SAL developed in this study could be a strategy to enhance the performance of SAL at the lung level. Graphical abstract.

How do people in China think about causes of their back pain? A predominantly qualitative cross-sectional survey.

BACKGROUND: Low back pain (LBP) is the second highest cause of health burden in China. Delayed recovery, poor clinical outcomes and persistence of LBP are associated with negative pain beliefs about LBP. Chinese philosophies are nested into the daily life of people in China, which is likely to influence pain beliefs. However, there is lack of knowledge about people's discourses regarding their LBP in China. The primary aim of this study was to explore the discourses underlying the beliefs of people in China about what causes their persistent or recurrent LBP. The secondary aim was to investigate the sources of these pain beliefs. METHODS: People (n = 152) from South Central, East and North Mainland China with LBP completed an online survey about what they believed caused their persistent or recurrent LBP and where these understandings came from. Potential causes of persistent or recurrent LBP were explored qualitatively using discourse analysis. The sources of these discourses were assessed by descriptive statistics with conventional content analysis. RESULTS: Five discourses were identified to underpin participants' beliefs about what caused their persistent or recurrent LBP, namely: (1) biomedical problems (66.4%), (2) unbalanced lifestyle (48.7%), (3) menstruation and 'kidney' status (9.2%), (4) the 'Five Elements' imbalance (7.9%), and (5) energy status (5.9%). Most participants responded that their pain beliefs were based on information derived from healthcare professionals (59.2%), followed by the internet (24.3%) and family (23.0%). CONCLUSIONS: People from moderately and well-developed parts of Mainland China think predominantly in line with a Western biomedical viewpoint about their LBP. Traditional Chinese medicine related pain beliefs mainly to the concept of 'balance' were evident on contemporary Chinese society's understandings of LBP. These cultural beliefs could be relevant to consider in LBP management and involve healthcare professionals, family and patient in this process.

Selective and sensitive determination of celastrol in traditional Chinese medicine based on molecularly imprinted polymers modified Mn-doped ZnS quantum dots optosensing materials.

In this work, we proposed a facile strategy to prepare molecularly imprinted polymers (MIPs) modified Mn-doped ZnS quantum dots (QDs) as optosensing materials via sol-gel polymerization for specific recognition of celastrol (Cel) in traditional Chinese medicine (TCM). Firstly, L-Cysteine (L-Cys) modified Mn-doped ZnS QDs (L-Cys@Mn-ZnS) was used as imprinting substrate. The amino and carboxyl groups on the surface of Mn-ZnS QDs can provide more binding sites for imprinting polymerization. Then, the fluorescent MIPs was synthesized in the presence of L-Cys@Mn-ZnS QDs, template celastrol, 3-aminopropyl triethoxysilane (APTES) and ammonium hydroxide in the ethanol-water (9/1, v/v) solution. The morphology and structure of the products were characterized by transmission electron microscopy (TEM), Fourier transform infrared spectroscopy (FT-IR), X-ray diffractometer (XRD) and X-ray photoelectron spectroscopy (XPS). The resulting MIPs functionalized Mn-doped ZnS QDs (denoted as MIPs@L-Cys@Mn-ZnS QDs) had higher imprinting factor of 14.19 and significant selectivity. The MIPs@L-Cys@Mn-ZnS QDs as fluorescent probe exhibited sensitive response to Cel in the linear range from 0.1 µM to 3.5 µM and the limit of detection was estimated to be 35.2 nM. The probe was also applied for the detection of Cel in traditional Chinese medicine with recovery ranged from 88.0%-105.0%. The results confirmed that MIPs@L-Cys@Mn-ZnS QDs could efficiently and specifically capture Cel from actual complex traditional Chinese medicine samples.

Pretreatment of Root Canal with Photodynamic Therapy Facilitates Adhesion, Viability and Differentiation of Stem Cells of the Apical Papilla.

This study was to test the hypothesis that root canal pretreated with photodynamic therapy (PDT) would promote stem cells from the apical papilla (SCAP) adhesion, proliferation and differentiation without affecting smear layer removal and microhardness of root canal. Standardized root canals were randomized into four groups (n = 30/group): (1) sodium hypochlorite(NaOCl) group, (2) NaOCl + ethylene diaminetetraacetic acid (EDTA) group, (3) NaOCl + PDT group, (4) NaOCl + EDTA + PDT group. After treatments, smear layer removal and microhardness of root canal were evaluated. SCAP with hydroxyapatite-based scaffolds were seeded into root canals for 7 days. SCAP adhesion was observed by scanning electron microscope (SEM), and viable cells were calculated by CellTiter-Glo Luminescent kit. Platelet-derived growth factor (PDGF) and vascular endothelial growth factor (VEGF) expression of SCAP were evaluated by Quantitative Reverse Transcriptase-Polymerase Chain Reaction. There was no significant difference in the smear layer removal and microhardness of root dentin between the groups with and without PDT treatment (P > 0.05). SCAP with elongated cytoplasmic processes and cell-cell contact were observed on the dentin surfaces treated with PDT. Elevated cell viability, PDGF and VEGF expression were found in root canal treated with PDT (P < 0.05). Under the experimental conditions, PDT could provide positive microenvironment for SCAP growth.

Preparation of magnetic molecularly imprinted polymers functionalized carbon nanotubes for highly selective removal of aristolochic acid.

In this work, we presented the preparation of magnetic carbon nanotubes (MCNTs) functionalized with molecularly imprinted polymers (MIPs) for effective removal of aristolochic acid I (AAI) in traditional Chinese medicine (TCM). MCNTs@AAI-MIPs was obtained via a facile and environmental friendly sol-gel process. Firstly, MCNTs were synthesized by a solvothermal method. Then, the template molecules were self-assembled with the functional monomer phenyltrimethoxysilane (PTMOS) in the presence of ethanol and water. Finally, AAI-MIPs film was coated on the MCNTs to obtain product MCNTs@AAI-MIPs using tetraethyl-orthosilicate (TEOS) as cross-linker. The morphology and structure of prepared MIPs were characterized by Fourier transform infrared spectroscopy (FT-IR), transmission electron microscopy (TEM), X-ray diffraction (XRD), nitrogen gas adsorption and vibrating sample magnetometer (VSM). The adsorption properties were demonstrated by kinetic, isothermal and selective adsorption experiments. The results showed that the imprinted nanocomposites exhibited fast separation rate (10 s), high adsorption capacity (18.54 µg∙mg-1), short kinetic equilibrium time (15 min), and good selectivity to template molecule with imprinting factor (IF) of 3.17. A regression equation (y=57294x-4734.1) with good linearity was obtained in the concentration range of 0.1-200 µg∙mg-1 for AAI with a correlation coefficient (R2) of 0.9998. The limit of detection (LOD, S/N=3) was 0.034 µg∙mg-1. Moreover, high recoveries ranged from 80% to 110% (RSD=3.27%-8.16%) were received in spiked TCM samples. The results suggested that the proposed MCNTs@AAI-MIPs could efficiently and specifically capture AAI from an actual complex TCM samples.

[Determination of 21 organophosphorus pesticides in tea by gas chromatography-mass spectrometry coupled with hydroxylated multi-walled carbon nanotubes based on dispersive solid-phase extraction].

A rapid determination method of 21 organophosphorus pesticides in tea was developed by QuEChERS method using modified multi-walled carbon nanotubes (MWCNTs-OH), primary-secondary amine (PSA) and MgSO4 coupled with gas chromatography-mass spectrometry. The pesticide residues in tea were extracted with a hexane-acetone (2:1, v/v) mixture, and cleaned up by dispersive solid-phase extraction using MWCNTs-OH and primary-secondary amine (PSA) as the sorbents. After centrifugation and filtration, the target compounds were analyzed by gas chromatography-mass spectrometry and quantified by the external standard method. Under the optimized conditions, good linearities were obtained in the range of 0. 01- 0. 50 mg/kg. The average recoveries were in the range of 81. 5% -109. 4% at three spiked levels, with relative standard deviations (RSDs, n = 5 ) of 2. 3% - 10. 6%. The limits of quantification were 0. 001-0. 040 mg/kg. This method is simple, fast, sensitive, cheap, and can meet the requirements of the rapid detection of organophosphorus pesticides in tea.

[Preparation and characterization of salvianolic acids-tanshinones composite powder for inhalation via co-micronized method].

Salvianolic acids and tanshinones are main hydrophilic and lipophilic extracts from Salvia Miltiorrhiza with significant anti-pulmonary fibrosis effects. The aim of this study was to prepare a co-micronized salvianolic acids-tanshinones composite powder for inhalation using a planetary ball mill. The micronization process parameters were optimized by central composite design (CCD) and response surface methodology (RSM). Treatment time, rotation speed and the ball/sample weight ratio were selected as the independent variables, and the volume fraction of particle size in 1-5 µm was taken as the dependent variable. The powder properties were evaluated by scanning electron microscopy (SEM), laser diffraction and X-ray powder diffraction (XRPD). The powder flow and hygroscopicity were determined with repose angle, compressibility index and critical relative humidity(CRH). According to the results, the salvianolic acids-tanshinones composite powder produced in optimal conditions had a narrow and unimodal particle size distribution and a smaller D50 of 2.33 µm. The volume fraction of particle size in 1-5 µm was 80.82%. The repose angle was (50.60±1.13) °, and the critical relative humidity is about 77%. After being micronized, the particle size significantly reduced, and the number of amorphous substances slightly increased, with no significant changes in powder flow and hygroscopicity. These findings indicate that the grinding method with a planetary ball mill can be used to co-micronize various components with different properties and prepare composite drug powders for dry powder inhalation.