Glutathione degradable manganese-doped polydopamine nanoparticles for photothermal therapy and cGAS-STING activated immunotherapy of lung tumor.

Photothermal therapy (PTT), which utilizes nanomaterials to harvest laser energy and convert it into heat to ablate tumor cells, has been rapidly developed for lung tumor treatment, but most of the PTT-related nanomaterials are not degradable, and the immune response associated with PTT is unclear, which leads to unsatisfactory results of the actual PTT. Herein, we rationally designed and prepared a manganese ion-doped polydopamine nanomaterial (MnPDA) for immune-activated PTT with high efficiency. Firstly, MnPDA exhibited 57.2% photothermal conversion efficiency to accomplish high-efficiency PTT, and secondly, MnPDA can be stimulated by glutathione (GSH) to the release of Mn2+, and it can produce ·OH in a Fenton-like reaction with the overexpressed H2O2 and stimulate the cyclic GMP-AMP synthase-stimulator of interferon genes (cGAS-STING) pathway. These two synergistically can effectively remove lung tumor cells that have not been ablated by PTT, resulting in an 86.7% tumor suppression rate under laser irradiation of MnPDA in vivo, and further significantly activated the downstream immune response, as evidenced by an increased ratio of cytotoxic T cells to immunosuppressive Treg cells. Conclusively, the GSH degradable MnPDA nanoparticles can be used for photothermal therapy and cGAS-STING-activated immunotherapy of lung tumors, which provides a new idea and strategy for the future treatment of lung tumors.

MAP7 promotes proliferation and migration of breast cancer cells and reduces the sensitivity of breast cancer cells to paclitaxel.

Breast cancer is a common malignancy that severely threatens women's mental health and lives. The paclitaxel-resistant breast cancer cells were established through a continuous stimulation with paclitaxel in a stepwise escalating concentration manner. The expression of MAP7 was detected by RT-P CR and western blot. The annexin V staining assay was used to measure the cell apoptosis ratio. The expression of cell invasive ability and apoptosis-related proteins was detected by western blot assay. The cellular motility was tested via transwell and wound healing assays. This study indicated that the MAP7 expression was upregulated in breast cancer cells and paclitaxel-resistant breast cancer cells. Moreover, downregulating MAP7 not only suppressed cell viability, motility and invasion, but also enhanced cellular apoptosis in paclitaxel-resistant breast cancer cells. In summary, this study investigated the effect of MAP7 protein on cell critical physiological function, which provided a novel potential target for treating paclitaxel-resistant breast cancer.

MRI Using Artificial Intelligence Algorithm to Evaluate Concurrent Chemoradiotherapy for Local Recurrence and Distant Metastasis of Cervical Squamous Cell Carcinoma.

The aim of this study was to investigate the magnetic resonance imaging (MRI) features of patients with local recurrence and distant metastasis of cervical squamous cell carcinoma before and after concurrent chemoradiotherapy based on artificial intelligence algorithm. In this study, 100 patients with cervical squamous cell carcinoma with local recurrence and distant metastasis who underwent concurrent chemoradiotherapy were collected as the research subjects, and all underwent MRI multisequence imaging scans. At the same time, according to the evaluation criteria of solid tumor efficacy, patients with complete remission were classified into the effective group, and patients with partial remission, progressive disease, and stable disease were classified into the ineffective group. In addition, an image segmentation algorithm based on Balloon Snake model was proposed for MRI image processing, and simulation experiments were carried out. The results showed that the Dice coefficient of the proposed model segmentation of the reconstructed image was significantly higher than that of the level set model and the greedy algorithm, while the running time was the opposite (P < 0.05). The lesion volume (38.76 ± 5.34 cm3) in the effective group after treatment was significantly smaller than that in the noneffective group (46.33 ± 4.64 cm3), and the rate of lesion volume shrinkage (28.71%) was significantly larger than that in the noneffective group (12.49%) (P < 0.05). The relative apparent diffusion coefficient (rADC) value and rADC value change rate of the lesion after treatment in the effective group were significantly greater than those in the noneffective group (P < 0.05). In summary, the image segmentation and reconstruction algorithm based on Balloon Snake model can not only improve the quality of MRI images but also shorten the processing time and improve the diagnostic efficiency. The volume regression rate and rADC value change rate of cervical squamous cell carcinoma lesion can reflect the early efficacy of concurrent chemoradiotherapy for cervical squamous cell carcinoma and have predictive value.

MicroRNA-1275 induces radiosensitization in oesophageal cancer by regulating epithelial-to-mesenchymal transition via Wnt/ß-catenin pathway.

Acquired radioresistance is one of the main obstacles for the anti-tumour efficacy of radiotherapy in oesophageal cancer (EC). Recent studies have proposed microRNAs (miRNAs) as important participators in the development of radioresistance in various cancers. Here, we investigated the role of miR-1275 in acquired radioresistance and epithelial-mesenchymal transition (EMT) in EC. Firstly, a radioresistant cell line KYSE-150R was established, with an interesting discovery was observed that miR-1275 was down-regulated in KYSE-150R cells compared to the parental cells. Functionally, miR-1275 inhibition elevated radioresistance in KYSE-150 cells via promoting EMT, whereas enforced expression of miR-1275 increased radiosensitivity in KYSE-150R cells by inhibiting EMT. Mechanically, we demonstrated that miR-1275 directly targeted WNT1 and therefore inactivated Wnt/ß-catenin signalling pathway in EC cells. Furthermore, WNT1 depletion countervailed the promoting effect of miR-1275 suppression on KYSE-150 cell radioresistance through hampering EMT, whereas WNT1 overexpression rescued miR-1275 up-regulation-impaired EMT to reduce the sensitivity of KYSE-150R cells to radiation. Collectively, our findings suggested that miR-1275 suppressed EMT to encourage radiosensitivity in EC cells via targeting WNT1-activated Wnt/ß-catenin signalling, providing a new therapeutic outlet for overcoming radioresistance of patients with EC.

Ionic Liquid-based Hollow Fiber Liquid-Liquid-Liquid Microextraction Combined with Capillary Electrophoresis for the Determination of Sulfonamides in Aquaculture Waters.

Ionic liquid-based hollow-fiber liquid-liquid-liquid microextraction (IL-HF-LLLME) coupled to capillary electrophoresis (CE) has been developed for the determination of six sulfonamides (SAs) in aquaculture waters. A series of extraction parameters was optimized to enhance the extraction efficiency, which included type and pore size of hollow fiber, type and composition of extraction solvent, pH value of donor phase, the concentration of acceptor phase and the mass ratio of donor phase to acceptor phase along with extraction temperature and time. Under optimal conditions, the IL-HF-LLLME-CE method provided a wide liner range for six SAs from 2 to 1,000 µg L-1 (r2 ≥ 0.9995), the limits of the detection from 0.25 to 0.48 and the enrichment factors from 122 to 230, respectively. Relative standard deviations for intra- and interday precision were 1.4-5.3% and 1.8-7.5% (n = 5), respectively. The proposed method was successfully applied for the determination of trace-level SAs in seven real-world aquaculture water samples with good recoveries (80.4-100.7%). Also, sulfamerazine and sulfamethoxazole were detected at the level of 0.52-1.60 µg L-1 in two water samples. Due to its good sensitivity, simple operation, short analysis time and eco-friendliness, the developed method has a great application potential in analysis of trace SA residues in aquaculture waters.

Effect and molecular mechanism of Lin28 on 5-Fu sensitivity of hepatocellular carcinoma HepG2 cells / 中国肿瘤生物治疗杂志

@# Objective: To investigate the effect and mechanism of RNA binding protein Lin28 on the 5-fluorouracil (5-Fu) sensitivity of HepG2 cells. Methods: HepG2 cells were transfected with plasmid pcDNA3.1-Lin28 or si-Lin28 (small interfering RNA of Lin28). qPCR and Western blotting were used to detect the expression of Lin28 in HepG2 cells after transfection. Changes of cell proliferation in transfected cells after 5-Fu treatment was detected by CCK8 assay and the 50% inhibitory concentration (IC50) was calculated. Flow cytometry was used to detect apoptotic rate after 5-Fu treatment and the expression of apoptosis-related protein was assayed by Western blotting. The mRNA expressions of drug-resistant miRNAs (let-7a and let-7b), as well as cancer stem cell markers (Oct4, Nanog and Sox2) after transfection were detected by qPCR. Results: As compared to the HepG2/Vector cells, the mRNA and protein expressions of Lin28 were significantly up-regulated in HepG2/Lin28 cells (P<0.05 or P<0.01). Over-expression of Lin28 significantly suppressed the sensitivity of HepG2 cells to 5-Fu (IC50elevated obviously, P<0.05) and significantly increased cell proliferation while decreased apoptotic rate and expression of apoptotic-related protein caspase-3 (all P<0.01). As compared to si-control group, expression of Lin28 in HepG2/si-Lin28 cells was significantly down-regulated (P<0.01). Lin28 knockdown significantly reduced cell proliferation and IC50 of 5-Fu (all P<0.01) but increased apoptotic rate and expression of apoptosis-related protein (P<0.01). Compared with HepG2/Vector group, expressions of let-7a and let-7b, as well as cancer stem cell markers (Oct4, Nanog and Sox2) were significantly increased in HepG2/Lin28 cells (all P<0.01); while these molecules were significantly decreased in HepG2/si-Lin28 cells as comparing to si-control group (all P<0.01). Conclusion: Lin28 can modulate the chemosensitivity of HepG2 cells by regulating the expression of miRNAs and the formation of cancer stem cells. Targeting Lin28 might be a promising approach to improve the chemotherapy efficacy in HCC.

Determination of aflatoxins in milk sample with ionic liquid modified magnetic zeolitic imidazolate frameworks.

The ionic liquid (IL) was introduced to the synthesis system of magnetic zeolite imidazolate framework-8 (M/ZIF-8), which was benefit to the formation of binary imidazole and the co-modification of M/ZIF-8. The morphology and textural properties of ILM/ZIF-8 were characterized by SEM, TEM, BET and BJH. The crystal structural shape and size of MZIF-8 was unvaried with the interventional of IL. The ILM/ZIF-8 was applied to the concentration and determination of aflaoxins (AFB1, AFB2, AFG1 and AFG2) in milk samples based on magnetic solid phase extraction (MSPE) coupled with ultra high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS). The experimental parameters of the MSPE, including amount of ILM/ZIF-8, pH, type and amount of desorption solvent, extraction time and sample volume were investigated by a univariate method and orthogonal screening. The four AFs were concentrated from the 20 mL milk when 90 mg ILM/ZIF-8 was used as magnetic adsorbent. The extraction efficiency of AFs was higher than 80.0% within 15 min. The limits of quantitative and detection were 7.5-26.7 and 2.3-8.1 ng/L, respectively. The proposed method was applied to the determination of milk samples containing trace amounts of AFs and the recoveries ranged from 79.0% to 102.5%, with RSD below 7.7%.

MicroRNA­301a targets WNT1 to suppress cell proliferation and migration and enhance radiosensitivity in esophageal cancer cells.

Esophageal cancer (EC) is one of the leading causes of death among malignancies. Radiotherapy for esophageal squamous cell carcinoma (ESCC) patients is limited by resistance to ionizing radiation (IR). An increasing body of evidence has demonstrated that aberrant expression of microRNA­301a (miR­301a) contributes to cancer progression and sensitivity to radiation. The aim of the present study was to investigate the exact functions and potential mechanisms of miR­301a in ESCC radioresistance. Initially, the miR­301a­transfected radioresistant ESCC cells KYSE­150R exhibited a decreased proliferation rate, and enhanced radiosensitivity and migration, whereas downregulation of miR­301a in radiosensitive KYSE­150 cells produced the opposite results. miR­301a regulates WNT1 expression at both the mRNA and protein levels. Furthermore, dual­luciferase reporter assays revealed that WNT1 was a target gene of miR­301a. In addition, the expression of miR­301a markedly affected the expression of Wnt/ß­catenin­related proteins such as ß­catenin and cyclin D1. Finally, overexpression of miR­301a inhibited epithelial­mesenchymal transition (EMT) conversion by directly targeting Snail and vimentin in radioresistant­ESCC cell lines; however, no inhibitory effects were exerted on Twist. Collectively, these results indicated that miR­301a increased the radiosensitivity and inhibited the migration of radioresistant­ESCC cells by targeting WNT1, thereby inactivating the Wnt/ß­catenin signaling pathway and EMT reversal. Thus, miR­301a may be a potential therapeutic target for the treatment of EC radioresistance.

Relationship between somatostatin receptor subtype expression and clinicopathology, Ki-67, Bcl-2 and p53 in colorectal cancer.

AIM: To study the SSTR1, 2, 3, 4, 5 expression and their relationships with clinico-pathological factors, cell proliferation, Bcl-2 and p53 expression in colorectal cancer cells. METHODS: Immunohistochemical staining of five SSTR subtypes, Ki-67, Bcl-2 and p53 was performed by the standard streptavidin-peroxidase (SP) technique for the paraffin sections of 127 colorectal cancers. and expression of five SSTR subtypes in 40 specimens of normal colorectal mucosae was detected with the same method. RESULTS: Positive staining for five SSTR subtypes was observed in colorectal cancer cells and normal colorectal mucosae. SSTR1 was the most predominant subtype in both colorectal cancer and normal colorectal mucosa, and the second was SSTR5 or SSTR2. As compared with normal colorectal mucosa, SSTR4 was more frequently expressed in colorectal cancer cells (2.5% vs 18.9%, P<0.05); the expression of SSTR2, 4, 5 in moderately to well differentiated colorectal adenocarcinoma was significantly higher than that in poorly differentiated ones (P<0.05), the SSTR1 expression in colorectal cancer with positive lymph node metastasis was significantly higher than that with negative lymph node metastasis (72.2% and 54.5%,P<0.05). In addition, in the ulcerative type of colorectal cancer, SSTR2 expression was obviously decreased (P<0.05); the correlation did not reach a statistical significance between the five SSTR subtypes expression and Dukes'stages (P>0.05), but the frequency of SSTR1 expression increased with Dukes'stage, while SSTR3 and SSTR5 expression decreased with Dukes' stage. Moreover, there was no correlation between expression of the five SSTR subtypes and other clinicopathological factors such as age, sex, tumor site, tumor depth, distant metastasis. The proliferative indexes in colorectal cancer cells with negative expression of SSTR2 and SSTR3 were significantly higher than that with positive expression (P<0.05). The Bcl-2 expression in colorectal cancer cells with positive expression of SSTR1, 2, 3, 5 was significantly lower than that with negative expression (P<0.05). There was no correlation between five SSTR subtypes and p53 expression. CONCLUSION: The most predominant SSTR subtype is SSTR1, and the second is SSTR2 or SSTR5. Five SSTR subtypes play different roles in the development of colorectal cancer. SSTR2 and SSTR3 can inhibit the proliferation and promote apoptosis of tumor cells.

Determination of paralytic shellfish toxins in dinoflagellate Alexandrium tamarense by using isotachophoresis/capillary electrophoresis.

Baseline separation of seven paralytic shellfish toxins (PSTs), namely decarbamoylsaxitoxin (dcSTX), saxitoxin (STX), neosaxitoxin (NEO), gonyautoxin-2 (GTX-2), gonyautoxin-3 (GTX-3), gonyautoxin-1 (GTX-1), and gonyautoxin-4 (GTX-4), was achieved by using capillary ITP (CITP)/CE with UV detection. Separation parameters including duration time and voltage in CITP process, separation voltage, and pH and concentration of buffer were optimized. The developed method provided linear responses from 1.3 to 200 microM for the PSTs. The LOD ranged from 0.1 to 0.3 microM. PST extracts from two algal strains of Alexandrium tamarense were analyzed and the toxin concentrations in the samples were quantified with an internal standard method by using NEO as the internal standard. The algal extract of A. tamarense HK9301 contained 332 microM GTX-2 and 224 microM GTX-3, while the PSTs were not detected in the extract of A. tamarense CI01.

Determination of phenol in landfill leachate by using microchip capillary electrophoresis with end-channel amperometric detection.

Phenol, 2,4-dichlorophenol (2,4-DCP), and 2,4,6-trichlorophenol (2,4,6-TCP) were baseline separated by using a homemade microchip CE with an end-channel amperometric detector where a 50 microm Pt microdisk working electrode (WE) and a Pt cathode were integrated onto the microchip itself. Separation parameters such as injection time and voltage, pH of the buffer, online pretreatment condition for WE, reproducibility, and detection potential were investigated. Under the selected separation conditions, the linear ranges for phenol, 2,4-DCP, and 2,4,6-TCP were 2-200, 4-400, and 4-400 microM, respectively. The LODs were 0.4, 0.5, and 0.7 microM for phenol, 2,4-DCP, and 2,4,6-TCP, respectively (S/N = 3). The standard addition method was successfully applied to the analysis of landfill leachate samples and the concentration of phenol in the landfill leachate samples was measured to be 0.32 and 0.21 mM, respectively. The recoveries were in the range of 85-103% and corresponding RSDs were less than 5.5%.

An end-channel amperometric detector for microchip capillary electrophoresis.

An end-channel amperometric detector with a guide tube for working electrode was designed and integrated on a home-made glass microchip. The guide tube was directly patterned and fabricated at the end of the detection reservoir, which made the fixation and alignment of working electrode relatively easy. The fabrication was carried out in a two-step etching process. A 30mum carbon fiber microdisk electrode and Pt cathode were also integrated onto the amperometric detector. The characteristics and primary performance of the home-made microchip capillary electrophoresis (MCCE) were investigated with neurotransmitters. The baseline separation of dopamine (DA), catechol (CA) and epinephrine (EP) was achieved within 80s. Separation parameters such as injection time, buffer components, pH of the buffer were studied. Relative standard deviations of not more than 6.0% were obtained for both peak currents and migration times. Under the selected separation conditions, the response for DA was linear from 5 to 200muM and from 20 to 800muM for CA. The limits of detection of DA and CA were 0.51 and 2.9muM, respectively (S/N=3).