2,5-Thiophenedicarboxylic Acid Bridging Hexameric CeIII-Substituted Selenotungstate and Its Application for Detecting Mucin 1.

The development of polyoxometalate chemistry not only is derived from the continuous discovery of novel polyoxometalates (POMs) but also stems from the exploitation of their new functionalities. In this work, we obtained a rigid sulfur-containing heterocyclic ligand-linking aggregate [N(CH3)4]10Na6H6[Ce8(H2O)26W8(HTDA)2(TDA)2O20][SeW4O18]2[SeW9O33]4·112H2O (1) (H2TDA = 2,5-thiophenedicarboxylic acid). Its polyanionic unit consists of one [Ce4(H2O)13W4O10(HTDA)(TDA)O10]18+ cluster and two kinds of Keggin-type [SeW4O18] and [SeW9O33] segments. It is noteworthy that H2TDA ligands not only work as connectors to link two symmetrical {[Ce4(H2O)13W4(HTDA)(TDA)O10][SeW4O18][SeW9O33]2}11- units but also function as ornaments to graft to the polyanionic backbone. Furthermore, 1 and 3,4-ethylenedioxythiophene (EDOT) were deposited on the glassy carbon electrode (GCE) by the electropolymerization (EPM) method, resulting in a 1-poly(3,4-ethylenedioxythiophene) (1-PEDOT) composite film, which can provide sufficient binding sites to immobilize Au nanoparticles (Au NPs). Hereafter, the Au NPs-immobilized 1-PEDOT modified electrode (Au/1-PEDOT/GCE) was used to construct an electrochemical aptasensor to detect mucin 1, showing a low detection limit of 29.5 fM in the Tris solution. This work not only demonstrates that rigid heterocyclic ligands are beneficial for the creation of novel rare-earth-substituted selenotungstate hybrids but also provides more enlightenment for POM-based materials used for electrochemical detection of cancer markers.

Mechanistic study on the promotion of Ca2+ leaching in steel slag through high-temperature solid waste modification.

Indirect carbonation of steel slag is an effective method for CO2 storage, reducing emissions, and promoting cleaner production in the steel industry. However, challenges remain, such as low Ca2+ leaching rates and slag management complexities arising from variations in mineral compositions. To address this, a high-temperature modification process is proposed to alter the mineral composition and facilitate the synergistic utilization of calcium and iron. This study delves into the effects of various solid waste modifications on the leaching of Ca2+ and the total iron content within steel slag. Results show that high-basicity modified slag forms Ca2(Al, Fe)2O5, reducing calcium leaching. Low-alkalinity modified slag produces calcium-rich aluminum minerals and also reduces the leaching of Ca2+ ions. At a basicity of 2.5, coal gangue, fly ash, and blast slag achieve maximum Ca2+ leaching rates of 88.93%, 89.46%, and 90.17%, respectively, with corresponding total iron contents of 41.46%, 37.72%, and 35.29%. Upgraded coal gangue exhibits a 50.02% increase in calcium leaching and a 15.58% increase in total iron content compared to the original slag. This enhances CO2 fixation and iron resource utilization. Overall, the proposed indirect carbonation and iron enrichment modification offer a novel approach for the resource utilization and environmental stability of steel slag.

Cytomodulin-10 modified GelMA hydrogel with kartogenin for in-situ osteochondral regeneration.

The incidence of osteochondral defect is increasing year by year, but there is still no widely accepted method for repairing the defect. Hydrogels loaded with bioactive molecules have provided promising alternatives for in-situ osteochondral regeneration. Kartogenin (KGN) is an effective and steady small molecule with the function of cartilage regeneration and protection which can be further boosted by TGF-ß. However, the high cost, instability, and immunogenicity of TGF-ß would limit its combined effect with KGN in clinical application. In this study, a composite hydrogel CM-KGN@GelMA, which contained TGF-ß1 analog short peptide cytomodulin-10 (CM-10) and KGN, was fabricated. The results indicated that CM-10 modified on GelMA hydrogels exerted an equivalent role in enhancing chondrogenesis as TGF-ß1, and this effect was also boosted when combined with KGN. Moreover, it was revealed that CM-10 and KGN had a synergistic effect on promoting the chondrogenesis of BMSCs by up-regulating the expression of RUNX1 and SOX9 at both mRNA and protein levels in vitro. Finally, the composite hydrogel exhibited a satisfactory osteochondral defect repair effect in vivo, showing similar structures close to the native tissue. Taken together, this study has revealed that CM-10 may serve as an alternative for TGF-ß1 and can collaborate with KGN to accelerate chondrogenesis, which suggests that the fabricated CM-KGN@GelMA composite hydrogel can be acted as a potential scaffold for osteochondral defect regeneration. STATEMENT OF SIGNIFICANCE: Kartogenin and TGF-ß have shown great value in promoting osteochondral defect regeneration, and their combined application can enhance the effect and show great potential for clinical application. Herein, a functional CM-KGN@GelMA hydrogel was fabricated, which was composed of TGF-ß1 mimicking peptide CM-10 and KGN. CM-10 in hydrogel retained an activity like TGF-ß1 to facilitate BMSC chondrogenesis and exhibited boosting chondrogenesis by up-regulating RUNX1 and SOX9 when being co-applied with KGN. In vivo, the hydrogel promoted cartilage regeneration and subchondral bone reconstruction, showing similar structures as the native tissue, which might be vital in recovering the bio-function of cartilage. Thus, this study developed an effective scaffold and provided a promising way for osteochondral defect repair.

Multi-CeIII-Inserted Phospho(III)tungstate Containing Three Building Units with Prominent Peroxidase-Mimicking Activity for Detecting l-Cysteine.

An attractive isonicotinic acid-ornamented octa-CeIII-inserted phospho(III)tungstate [H2N(CH3)2]6Na8[Ce8(H2O)30W8Na2O20(INA)4][HPIIIW4O17]2[HPIIIW9O33]4·30H2O (1-Ce) (HINA = isonicotinic acid) has been isolated through the deliberately designed one-step assembly strategy, in which the HPO32- heteroanion template was introduced into the Ce3+/WO42- system in the presence of HINA. The polyoxoanion of 1-Ce consists of two identical [Ce4(H2O)15W4NaO10(INA)2][HPIIIW4O17][HPIIIW9O33]2}7- subunits linked by Ce-O-W bonds. The polyoxoanion exhibits three kinds of polyoxotungstate building blocks [W4NaO20(INA)2]17-, [HPIIIW4O17]6-, and [HPIIIW9O33]8-, in which [W4NaO20(INA)2]17- and [HPIIIW4O17]6- building units can be considered as seeds driven by the coordination of additional Ce3+ ions to induce aggregation of [HPIIIW9O33]8- fragments. Furthermore, 1-Ce possesses high peroxidase-like activity and can oxidize 3,3',5,5'-tetramethylbenzidine in the presence of H2O2 with a turnover rate of 6.20 × 10-3 s-1. Because l-cysteine (l-Cys) can reduce oxTMB to TMB, the detection of l-Cys was established based on the 1-Ce-based H2O2 colorimetric biosensing platform with the linear range of 5-100 µM and the limit of detection of 4.28 µM. This work not only can expand the scientific research studies on coordination chemistry and materials chemistry of rare-earth-inserted polyoxotungstates but also can provide practical application possibility in clinical diagnosis using liquid biopsy.

Transcription factor c-Jun modulates GLUT1 in glycolysis and breast cancer metastasis.

As the main isoforms of membranous glucose transporters (GLUT), GLUT1 involves tumorigenesis, metastasis and prognosis in a variety of cancers. However, its role in breast cancer metastasis remains to be elucidated. Here we examined its transcriptional and survival data in patients with breast cancer from several independent databases including the Oncomine, Gene Expression Profiling Interactive Analysis, Gene Expression across Normal and Tumor tissue, UALCAN, cBioPortal, Kaplan-Meier Plotter and PROGgeneV2. We found that its mRNA expression was significantly high in cancer tissues, which was associated with metastasis and poor survival. Transcription factor c-Jun might bind to GLUT1 promoter to downregulate its gene expression or mRNA stability, therefore to suppress glycolysis and metastasis. By qRT-PCR, we verified that GLUT1 was significantly increased in 38 paired human breast cancer samples while JUN was decreased. Furthermore, the protein level of GLUT1 was higher in tumor than in normal tissues by IHC assay. To explore underlying pathways, we further performed GO and KEGG analysis of genes related to GLUT1 and JUN and found that GLUT1 was increased by transcription factor c-Jun in breast cancer tissues to influence glycolysis and breast cancer metastasis.

B7-H3 targeted CAR-T cells show highly efficient anti-tumor function against osteosarcoma both in vitro and in vivo.

BACKGROUND: Osteosarcoma (OS) mainly happens in children and youths. Surgery, radiotherapy and chemotherapy are the common therapies for osteosarcoma treatment but all their anti-tumor effects are limited. In recent years, a new cellular therapy, CAR-T, a cellular immunotherapy with genetically engineered T cells bearing chimeric antigen receptor targeting specific tumor-associated antigen, has been proved to be an effective therapy against acute lymphoblastic leukemia. Thus, CAR-T is a potentially effective therapy for osteosarcoma treatment. METHODS: A CAR gene targeting B7-H3 antigen was constructed into lentiviral vector through molecular biology techniques. Then, the CAR gene was transferred to T cells through lentiviral delivery system, and the CAR-T cells were largely expanded using in vitro culture technology. The in vitro anti-tumor effect of CAR-T cells was evaluated through Real Time Cell Analysis system (RTCA) and ELISA assay. The in vivo anti-tumor capabilities of CAR-T cells were evaluated using the patient-derived xenografts (PDX) model of osteosarcoma. RESULTS: The third-generation CAR-T cells we constructed could target the B7-H3 antigen, and the phenotype of CAR-T cells was consistent with normal T cells; The CAR-T cells showed superior antitumor effects both in vitro and in vivo. CONCLUSION: Our study showed that B7-H3 targeted CAR-T cells had high anti-tumor efficacy against osteosarcoma both in vitro and in vivo, which proved that B7-H3 targeted CAR-T therapy is potentially effective for osteosarcoma treatment.

Prognostic analysis of hepatocellular carcinoma with macrovascular invasion after liver resection and a successful case of conversion therapy.

Objective: Macrovascular invasion (MVI) is an important factor leading to poor prognosis in hepatocellular carcinoma (HCC). Liver resection may offer favorable prognosis for selected patients with HCC. This study aimed to analyze the prognostic factors of HCC with MVI after liver resection as well as demonstrate a case of conversion therapy in an HCC patient with portal vein tumor thrombus (PVTT). Methods: A total of 168 HCC patients with MVI who underwent primary liver resection at the Affiliated Hospital of Qingdao University between January 2013 and October 2021 were enrolled in the study. Clinicopathological data were collected retrospectively. Univariate and multivariate regression analyses were used to investigate the risk factors influencing recurrence and overall survival. Additionally, conversion therapy with drug-eluting bead transarterial chemoembolization (D-TACE), and sorafenib plus sintilimab treatment was performed in an HCC patient with PVTT. Results: Among the 168 patients with HCC, 11 were diagnosed with hepatic vein tumor thrombosis, and the rest were diagnosed with PVTT. The 1-year disease-free survival rate was 37.5%, and the 3-year overall survival rate was 52.7%. Univariate and multivariate regression analyses revealed that HBsAg positivity, alpha-fetoprotein (AFP) level ≥400 ng/ml, liver capsule invasion, and tumor number ≥2 were independent prognostic factors for tumor recurrence, whereas HBsAg positivity was an independent risk factor for overall survival. Postoperative prophylactic medication did not significantly prolong the recurrence time. The median survival time (MST) after tumor recurrence was 13.4 months. In the patient treated with conversion therapy, the tumor gradually shrank and was eventually surgically resected. Conclusions: This study identified the independent prognostic and risk factors associated with recurrence and overall survival in HCC patients with MVI. Additionally, we successfully performed conversion therapy in an HCC patient with PVTT. The findings would help identify patients at high risk of recurrence and indicate that combined therapy may prolong the survival of HCC patients with PVTT.

Ginsenoside Rc ameliorated atherosclerosis via regulating gut microbiota and fecal metabolites.

Atherosclerosis (AS) and the accompanied cardiovascular diseases (CVDs) were the leading cause of death worldwide. Recently, the association between CVDs, gut microbiota, and metabolites had aroused increasing attention. In the study, we headed our investigation into the underlying mechanism of ginsenoside Rc (GRc), an active ingredient of ginsenosides used for the treatment of CVDs, in apolipoprotein E-deficient (ApoE-/-) mice with high-fat diet (HFD). Seven-week-old male ApoE-/- mice were randomly divided into four groups: the normal control (NC) group, the HFD group, the GRc group (40 mg/kg/d), and the atorvastatin (Ato) group (10 mg/kg/d). Atherosclerotic injury was evaluated by aortic lesions, serum lipid levels, and inflammatory factors. The composition of gut microbiota and fecal metabolite profile were analyzed using 16S rRNA sequence and untargeted metabolomics, respectively. The results showed that GRc significantly alleviated HFD-induced aortic lesions, reduced serum levels of total cholesterol (TC), triglyceride (TG), low-density lipoprotein cholesterol (LDL-C), tumor necrosis factor-α (TNF-α), and interleukin (IL)-6 and IL-1ß, and increased high-density lipoprotein cholesterol (HFD-C) level, as well as the alteration of gut microbiota composition, function, and metabolite profile. GRc also reversed HFD change of Bacteroidetes and Firmicutes at the phylum level, Muribaculaceae, Lactobacillus, Ileibacterium, Bifidobacterium, Faecalibaculum, Oscillibacter, Blautia, and Eubacterium_coprostanoligenes_group at the genus level, and 23 key metabolites involved in taurine and hypotaurine metabolism, arginine biosynthesis, ATP-binding cassette (ABC) transporters, primary bile acid biosynthesis, purine metabolism, tricarboxylic acid (TCA) cycle, and glucagon signaling pathways. Additionally, eight differential intestinal floras at the genus level were associated with 23 key differential metabolites involving atherosclerotic injury. In conclusion, our results demonstrated that GRc ameliorated atherosclerotic injury, regulated microbial and metabolomic changes in HFD-induced ApoE-/- mice, and suggested a potential correlation among gut microbiota, metabolites, and atherosclerotic injury regarding the mechanisms of GRc against AS.

Nuciferine Regulates Immune Function and Gut Microbiota in DSS-Induced Ulcerative Colitis.

Nuciferine, a major aporphine alkaloid obtained from the leaves of Nelumbo nucifera, exhibits anti-cancer and anti-inflammatory properties; however, its protective effects against inflammatory bowel diseases (IBD) has never been explored. In this study, an ulcerative colitis (UC) model was established in BALb/c mice by the continuous administration of 5% dextran sulfate sodium (DSS) in drinking water for 1 week. From day 8 to day 14, the DSS-treated mice were divided into a high-dose and a low-dose nuciferine treatment group and were intraperitoneally injected with the corresponding dose of the drug. Body weight loss, disease activity index (DAI), and colon length were measured. Histological changes were observed using hematoxylin and eosin staining. T lymphocyte proliferation was assessed by MTT assay. The ratio of CD3+, CD4+, CD8+, Th1, Th2, Th17, and Treg cells were estimated by flow cytometry. Finally, 16S rRNA sequencing was performed to compare the composition and relative abundance of the gut microbiota among the different treatment groups. The results showed that nuciferine treatment led to a significant improvement in symptoms, such as histological injury and colon shortening in mice with DSS-induced UC. Nuciferine treatment improved the Th1/Th2 and Treg/Th17 balance in the DSS-induced IBD model, as well as the composition of the intestinal microflora. At the phylum level, compared with the control group, the abundance of Firmicutes and Actinobacteriota was decreased in the model group, whereas that of Bacteroidetes increased. Meanwhile, at the genus level, compared with the control group, the numbers of the genera Lachnospiraceae_Clostridium, Bilophila and Halomonas reduced in the model group, while those of Bacteroides, Parabacteroides, and Paraprevotella increased. Notably, nuciferine administration reversed this DSS-induced gut dysbiosis. These results indicated that nuciferine modulates gut microbiota homeostasis and immune function in mice with DSS-induced UC.

Transcription factors Bcl11a and Bcl11b are required for the production and differentiation of cortical projection neurons.

The generation and differentiation of cortical projection neurons are extensively regulated by interactive programs of transcriptional factors. Here, we report the cooperative functions of transcription factors Bcl11a and Bcl11b in regulating the development of cortical projection neurons. Among the cells derived from the cortical neural stem cells, Bcl11a is expressed in the progenitors and the projection neurons, while Bcl11b expression is restricted to the projection neurons. Using conditional knockout mice, we show that deficiency of Bcl11a leads to reduced proliferation and precocious differentiation of cortical progenitor cells, which is exacerbated when Bcl11b is simultaneously deleted. Besides defective neuronal production, the differentiation of cortical projection neurons is blocked in the absence of both Bcl11a and Bcl11b: Expression of both pan-cortical and subtype-specific genes is reduced or absent; axonal projections to the thalamus, hindbrain, spinal cord, and contralateral cortical hemisphere are reduced or absent. Furthermore, neurogenesis-to-gliogenesis switch is accelerated in the Bcl11a-CKO and Bcl11a/b-DCKO mice. Bcl11a likely regulates neurogenesis through repressing the Nr2f1 expression. These results demonstrate that Bcl11a and Bcl11b jointly play critical roles in the generation and differentiation of cortical projection neurons and in controlling the timing of neurogenesis-to-gliogenesis switch.

Heptadecanoic acid and pentadecanoic acid crosstalk with fecal-derived gut microbiota are potential non-invasive biomarkers for chronic atrophic gastritis.

Background: Chronic atrophic gastritis (CAG), premalignant lesions of gastric cancer (GC), greatly increases the risk of GC. Gastroscopy with tissue biopsy is the most commonly used technology for CAG diagnosis. However, due to the invasive nature, both ordinary gastroscope and painless gastroscope result in a certain degree of injury to the esophagus as well as inducing psychological pressure on patients. In addition, patients need fast for at least half a day and take laxatives. Methods: In this study, fecal metabolites and microbiota profiles were detected by metabolomics and 16S rRNA V4-V5 region sequencing. Results: Alteration of fecal metabolites and microbiota profiles was found in CAG patients, compared with healthy volunteers. To identify the most relevant features, 7 fecal metabolites and 4 microbiota were selected by random forest (RF), from A and B sample sets, respectively. Furthermore, we constructed support vector machines (SVM) classifification model using 7 fecal metabolites or 4 gut microbes, or 7 fecal metabolites with 4 gut microbes, respectively, on C sample set. The accuracy of classifification model was 0.714, 0.857, 0.857, respectively, and the AUC was 0.71, 0.88, 0.9, respectively. In C sample set, Spearman's rank correlation analysis demonstrated heptadecanoic acid and pentadecanoic acid were signifificantly negatively correlated to Erysipelotrichaceae_UCG-003 and Haemophilus, respectively. We constructed SVM classifification model using 2 correlated fecal metabolites and 2 correlated gut microbes on C sample set. The accuracy of classification model was 0.857, and the AUC was 0.88. Conclusion: Therefore, heptadecanoic acid and pentadecanoic acid, crosstalk with fecal-derived gut microbiota namely Erysipelotrichaceae_UCG-003 and Haemophilus, are potential non-invasive biomarkers for CAG diagnosis.

Disruption of adenosine 2A receptor improves the anti-tumor function of anti-mesothelin CAR T cells both in vitro and in vivo.

Chimeric antigen receptor (CAR) T cells have been successfully used for the treatment of hematological malignancies including acute and chronic lymphoblastic leukemia. However, results of CAR T cell projects in solid tumors have been less impressive to date, partly because of immunosuppressive tumor microenvironment (TME). It is widely known that high adenosine production is an important factor causing tumor-induced immunosuppression in TME, and adenosine mediates the suppression of anti-tumor T cell responses via binding and signaling through adenosine 2a receptor (A2aR). Previous studies have shown that adenosine generated by cancer cells significantly inhibits T cell anti-tumor activity through binding and then activating adenosine 2A receptors (A2aRs) of T cells. Based on the previous work, in our study, we evaluated whether A2aR disruption by shRNA could enhance the anti-tumor function of anti-mesothelin (MSLN) CAR T cells both in vitro and in vivo. For this goal above, we used MSLN-positive human ovarian serous carcinoma cells (SKOV3) and human colon cancer cells (HCT116) as target cancer cells while MSLN-negative human ovarian cancer cells (ES2) as non-target cancer cells. We observed that targeting cell-intrinsic A2aR through shRNA overexpression caused significant A2aR disruption in CAR T cells and profoundly increased CAR T cell efficacy in both CAR T cell cytokine production and cytotoxicity towards MSLN-positive cancer cells in vitro. More importantly, in SKOV3 xenograft mouse models, anti-MSLN CAR-T cells significantly reduced the tumor burden compared with non-transduced T cells, and the anti-tumor activity of A2aR-disrupted anti-MSLN CAR-T cells was stronger than that of wild-type anti-MSLN CAR-T cells. Altogether, our study showed enhanced anti-tumor efficacy caused by shRNA-mediated A2aR disruption in anti-MSLN CAR T cells both in vitro and in vivo, which proved that shRNA-mediated modification of gene expression might be an excellent strategy for improving CAR T cell function in immunosuppressive tumor microenvironment (TME) and could potentially improve the outcome of treatment in clinical trials.

Detection of residual E. coli host cell DNA by 23S ribosomal RNA gene-targeted quantitative polymerase chain reactions.

Among the many systems available for heterologous protein production gram-negative bacterium Escherichia coli (E. coli) has long been widely used because of its ability to grow rapidly with a high density on inexpensive substrates. The use of E. coli as the host system has many regulatory issues, one of which is the residual host cell DNA. Residual DNA carried by biological products may lead to carcinogenicity and immunomodulation risks. The World Health Organization (WHO) for the acceptable amounts of residual host cell DNA is less than 10 ng per dose. Therefore, it is important to keep an extremely low level of residual host DNA in the biological products derived from E. coli. In this study, we designed primer/probe sets targeting E. coli 23S ribosomal RNA gene to quantify the residual DNA of E. coli by quantitative polymerase chain reactions (qPCR). Result showed that this primer/probe has high species specificity. The limit of detection (LOD) in this method is 0.01 pg/µl and this allowed for detection of residual host DNA of much lower concentrations. We assessed accuracy by calculating the recovery (92.1∼140.1 %) of the spiked DNA in plasmids which were produced from E. coli. We also checked intra-assay precision (9.8∼15.1 %) and inter-assay precision (10.9∼18.3 %) by repeatedly measuring the four different concentration standards. In addition, the robustness assay was performed by generating standard curve using short length E. coli DNA. The result showed that appropriate degree of DNA fragmentation will not affect tests. These validation studies demonstrated that our method has excellent specificity, linearity, accuracy, precision and robustness.

Metabolic changes from exposure to harmful Maillard reaction products and high-fat diet on Sprague-Dawley rats.

Rats were fed a high-fat diet and treated with acrylamide (21 mg/kg BW), harmane (7 mg/kg BW), NƐ-(carboxymethyl)lysine (CML; 10 mg/kg BW), or the co-accumulation of the three chemicals to investigate the effects of harmful Maillard reaction products (MRPs) generated by heat processing of potatoes. Damage to the liver, kidney, gastrocnemius, and nervous system was confirmed by serum biochemical testing and histopathology examination after six weeks of feeding. Serum metabolomic analysis was performed using gas chromatography time-of-flight mass spectrometry (GC-TOF-MS). The metabolites detected by GC-TOF-MS were processed using chemometric techniques, and the results were evaluated using a t-test (p < 0.05) and calculated variable importance in projection (VIP) value (>1.5). Significant changes (increase or decrease) were identified in 13 metabolites, compared with the control. These metabolites were gluconolactone, tyrosol, 5-hydroxytryptophan, erythrose 4-phosphate, quinolinic acid, 2-ketobutyric acid, tyrosine, 3-hydroxy-3-methylglutaric acid, desaminotyrosine, L-cysteine, ribonic acid, cysteamine, and tryptophan. Among these, cysteamine and tryptophan are metabolites unique to the co-accumulation of all three test compounds in the treated rats. This study demonstrates that these harmful MRPs exert toxic effects on the liver, kidney, and nervous system via different metabolic pathways. Moreover, co-accumulation of these harmful MRPs caused differing toxic effects and changes in a variety of metabolites. These changes may contribute to the interaction of those harmful MRPs and also affect toxicity at the metabolic level.

The antitumor capacity of mesothelin-CAR-T cells in targeting solid tumors in mice.

Since the approval of chimeric antigen receptor (CAR) T cell therapy targeting CD19 by the FDA, CAR-T cell therapy has received increasing attention as a new method for targeting tumors. Although CAR-T cell therapy has a good effect against hematological malignancies, it has been less effective against solid tumors. In the present study, we selected mesothelin (MSLN/MESO) as a target for CAR-T cells because it is highly expressed by solid tumors but only expressed at low levels by normal tissues. We engineered a third generation MSLN-CAR comprising a single-chain variable fragment (scFv) targeting MSLN (MSLN-scFv), a CD8 transmembrane domain, the costimulatory domains from CD28 and 4-1BB, and the activating domain CD3ζ. In vitro, MSLN-CAR-T cells killed various solid tumor cell lines, demonstrating that it could specifically kill MSLN-positive cells and release cytokines. In vivo, we investigated the effects of MSLN-CAR-T cell therapy against ovarian, breast, and colorectal cancer cell-line-derived xenografts (CDX) and MSLN-positive colorectal and gastric cancer patient-derived xenografts (PDX). MSLN-CAR decreased the growth of MSLN-positive tumors concomitant with significantly increased T cells and cytokine levels compared to the control group. These results indicated that modified MSLN-CAR-T cells could be a promising therapeutic approach for solid tumors.

3-D Antimonotungstate Framework Based on 2,6-H2pdca-connecting Iron-Cerium Heterometallic Krebs-type Polyoxotungstates for Detecting Small Biomolecules.

An inorganic-organic hybrid 3-D FeIII-CeIII heterometallic antimonotungstate framework [Ce(H2O)5(2,6-pdca)]4H2[Fe4(H2O)6(SbW9O33)2]·38H2O (1) (2,6-H2pdca = 2,6-pyridine-dicarboxylic acid) has been synthesized via a hydrothermal method by the one-pot reaction of 2,6-H2pdca, FeCl3·6H2O, Ce(NO3)3·6H2O, and Na9[B-α-SbW9O33]·19.5H2O. Notably, the structural unit of 1 possesses a Krebs-type [Fe4(H2O)6(2,6-pdca)2(SbW9O33)2]10- subunit supported with four bridging [Ce(H2O)5(2,6-pdca)]+ moieties. It is worth highlighting that adjacent structural units are concatenated together through heterobimetallic bridges to construct a 3-D framework. Furthermore, cuboid nanocrystal 1' was prepared under mild hydrothermal conditions based on the electrostatic interaction between 1 and K+. The effects of concentration and time on the morphology of nanocrystal 1' were also studied. The cuboid nanocrystal 1' was used as a modified electrode material for simultaneous electrochemical detection of dopamine and acetaminophen. The 1'-modified glassy carbon electrode shows good selectivity and sensitivity for detecting dopamine and acetaminophen.

Effects of different food ingredients on the color and absorption spectrum of carminic acid and carminic aluminum lake.

In this study, three foodstuffs (surimi, minced meat, and milk) were dyed with carminic acid and carminic aluminum lake. The effects of protein, metal ions, and food additives on the color of carminic acid and carminic aluminum lake were investigated. After being dyed by carminic acid, the colors of surimi, minced meat, and milk were light purple, red, and gray-green, respectively. When using carminic aluminum lake, surimi and milk were magenta, and minced meat was red. Regarding the carminic acid solution, the presence of myofibrillar protein (MFP), whey protein isolate (WPI), and soy protein isolate (SPI) turned it red by changing the pH, while the presence of casein made it orange. The carminic aluminum lake solution turned magenta in all four cases, which were not affected by protein. The color of carminic acid and carminic aluminum lake was significantly affected by 0.001-0.1 mol/L Fe3+, 0.001-0.1 mol/L Fe2+, 0.001-0.1 mol/L Cu2+, and 0.1 mol/L Ca2+, limiting their application in iron-, copper-, and high-calcium foods. The color of carminic acid was changed to yellow by 0.01%-1% sodium nitrite, but 0.01%-1% ascorbic acid and 0.01%-0.1% monascus color did not significantly affect the color of either carminic acid or carminic aluminum lake. This paper provides a reference for the application of carminic acid and carminic aluminum lake in food science.

The Effect of Exogenous Free Nε-(Carboxymethyl)Lysine on Diabetic-Model Goto-Kakizaki Rats: Metabolomics Analysis in Serum and Urine.

The current study investigated the effects of exogenous free Nε-(carboxymethyl) lysine (CML) from daily diet on diabetic-model Goto-Kakizaki rats. Rats were fed with free CML (2 mg/kg body weight) for 8 weeks, then metabolomics evaluation was performed on serum and urine, and biochemical and histopathologic examinations were conducted to verify metabolic results. Diabetic rats fed with free CML showed significantly increased (P < 0.05) fasting blood glucose (11.1 ± 1.07 mmol/L) and homeostasis model assessment values (homeostatic model assessment of insulin resistance: 16.0 ± 4.24; homeostatic model assessment of beta cell function: 6.66 ± 2.01; and modified beta cell function index: 11.5 ± 2.66) and a significantly altered (P < 0.05) oxidative stress level when compared to the control group. Serum and urine metabolomics showed a significantly altered (P < 0.05) level of aminomalonic acid, 2-oxoadipic acid, l-malic acid, ß-alanine, 2-oxoglutaric acid, d-threitol, N-acetyl-leucine, methylmalonic acid, l-cysteine, thymine, glycine, l-alanine, 4-hydroxyproline, hexadecane, succinic acid, l-ornithine, gluconolactone, maleic acid, l-lactate, tryptophan, 5-methoxyindoleacetate, γ-aminobutyric acid, homoserine, maltose, and quinolinic acid. Our results indicated that these metabolites altered by exposure to exogenous free CML were mapped to the citric acid cycle and amino acid and carbohydrate metabolism, which might be related to increased progression of diabetes and some other diabetic complications, including diabetic brain and neurological diseases, retinopathy, nephropathy, and impaired wound healing.

[Treatment of lumbar tuberculosis with percutaneous pedicle screw internal fixation and minimally invasive lateral small incisions lesion debridement and bone graft fusion via dilated channels].

OBJECTIVE: To explore the efficacy of percutaneous pedicle screw internal fixation and minimally invasive lateral small incisions lesion debridement and bone graft fusion via dilated channels in the treatment of lumbar tuberculosis. METHODS: The clinical data of 22 cases of lumbar tuberculosis treated with percutaneous pedicle screw internal fixation combined with dilated channels with minimally invasive lateral small incision lesion debridement and bone graft fusion between January 2016 and June 2018 were retrospectively analyzed. There were 12 males and 10 females, with an average age of 47.5 years (range, 22-75 years). The affected segments were L 2, 3 in 5 cases, L 3, 4 in 8 cases, and L 4, 5 in 9 cases, with an average disease duration of 8.6 months (range, 4-14 months). Preoperative neurological function was classified according to the American Spinal Injury Association (ASIA), with 3 cases of grade C, 9 cases of grade D, and 10 cases of grade E. The operation time, intraoperative blood loss, and postoperative complications were recorded. At preoperation, 3 months after operation, and last follow-up, the C reactive protein (CRP) and erythrocyte sedimen- tation rate (ESR) were tested to evaluate tuberculosis control; the pain visual analogue scale (VAS) score was used to evaluate the recovery of pain, and the Oswestry disability index (ODI) was used to evaluate the function recovery of the patient's lower back; the kyphosis Cobb angle was measured, and the loss of Cobb angle (the difference between the Cobb angle at last follow-up and 3 months after operation) was calculated. At last follow-up, the ASIA classification was used to evaluate the recovery of neurological function, and the effectiveness was evaluated according to the modified MacNab standard. RESULTS: The operation time was 110-148 minutes (mean, 132.8 minutes) and the intraoperative blood loss was 70-110 mL (mean, 89.9 mL). Two patients experienced fat liquefaction of the incision and delayed healing; the incisions of the remaining patients healed by first intention. All patients were followed up 18-24 months, with an average of 21.3 months. All bone grafts achieved osseous fusion, the pedicle screws were fixed in reliable positions, without loosening, displacement, or broken rods. There was no recurrence of tuberculosis. The ESR, CRP, VAS scores, ODI scores, and kyphosis Cobb angle of the affected segment at 3 months after operation and last follow-up were significantly improved ( P<0.05); there were no significant differences between at last follow-up and 3 months after operation ( P>0.05), and the loss of Cobb angle was (0.6±0.5)°. The patient's neurological function recovered significantly. At last follow-up, the ASIA grades were classified into 1 case with grade C, 1 case with grade D, and 20 cases with grade E, which were significantly improved when compared with preoperative grading ( Z=-3.066, P=0.002). According to the modified MacNab standard, 16 cases were excellent, 3 cases were good, 2 cases were fair, and 1 case was poor. The excellent and good rate was 86.4%. CONCLUSION: Percutaneous pedicle screw internal fixation combined with dilated channels with minimally invasive lateral small incisions lesion debridement and bone graft fusion has the advantages of less bleeding, less trauma, and faster recovery, which is safe and effective in the treatment of lumbar tuberculosis.

PD-1 silencing improves anti-tumor activities of human mesothelin-targeted CAR T cells.

Chimeric antigen receptor T (CAR T) cell therapy is a new pillar in cancer therapeutics, and has been successfully used for the treatment of cancers, including acute lymphoblastic leukemia and solid cancers. Following immune attack, many tumors upregulate inhibitory ligands which bind to inhibitory receptors on T cells. For example, the interaction between programmed cell death protein 1 (PD-1) on activated T cells and its ligands (widely known as PD-L1) on a target tumor limits the efficacy of CAR T cells therapy against poorly responding tumors. Here, we use mesothelin (MSLN)-expressing human ovarian cancer cells (SKOV3) and human colon cancer cells (HCT116) to investigate whether PD-1-mediated T cell exhaustion affects the anti-tumor activity of MSLN-targeted CAR T cells. We utilized cell-intrinsic PD-1-targeting shRNA overexpression strategy, resulting in a significant PD-1 silencing in CAR T cells. The reduction of PD-1 expression on T cell surface strongly augmented CAR T cell cytokine production and cytotoxicity towards PD-L1-expressing cancer cells in vitro. This study indicates the enhanced anti-tumor efficacy of PD-1-silencing MSLN-targeted CAR T cells against several cancers and suggests the potential of other specific gene silencing on the immune checkpoints to enhance the CAR T cell therapies against human tumors.