Single dual-specific anti-PD-L1/TGF-ß antibody synergizes with chemotherapy as neoadjuvant treatment for pancreatic ductal adenocarcinoma: a preclinical experimental study.

AIMS: Chemotherapy resistance is an important cause of neoadjuvant therapy failure in pancreatic ductal adenocarcinoma (PDAC). BiTP (anti-PD-L1/TGF-ß bispecific antibody) is a single antibody that can simultaneously and dually target transforming growth factor-beta (TGF-ß) and programmed cell death ligand 1 (PD-L1). We attempted in this study to investigate the efficacy of BiTP in combination with first-line chemotherapy in PDAC. METHODS: Preclinical assessments of BiTP plus gemcitabine and nab-paclitaxel were completed through a resectable KPC mouse model (C57BL/6J). Spectral flow cytometry, tissue section staining, enzyme-linked immunosorbent assays, Counting Kit-8, transwell, and Western blot assays were used to investigate the synergistic effects. RESULTS: BiTP combinatorial chemotherapy in neoadjuvant settings significantly downstaged PDAC tumors, enhanced survival, and had a higher resectability for mice with PDAC. BiTP was high affinity binding to targets and reverse chemotherapy resistance of PDAC cells. The combination overcame immune evasion through reprogramming tumor microenvironment via increasing penetration and function of T cells, natural killer cells, and dendritic cells and decreasing the function of immunosuppression-related cells as regulatory T cells, M2 macrophages, myeloid-derived suppressor cells, and cancer-associated fibroblasts. CONCLUSION: Our results suggest that the BiTP combinatorial chemotherapy is a promising neoadjuvant therapy for PDAC.

Positive Synergy between the Helical Poly(phenylacetylene) Backbones and the Helical L-Proline Oligopeptide Pendants for Enhanced Enantioseparation Properties.

A series of optically active helical poly(phenylacetylene)s (PPA-Pro1, PPA-Pro3, PPA-Pro6, PPA-Pro9, and PPA-Pro12) bearing different chain lengths of L-proline oligopeptide in the side chains were obtained by polymerizing the corresponding novel phenylacetylene monomers. The monomer adopted a trans-rich helix structure when the L-proline oligopeptide chain length was longer, according to the optical activities and 2D-NMR analysis. The helical structure could be maintained and significantly influenced the polymers' helical conformation by introducing the L-proline oligopeptide to the pendants. By the way, the morphology of PPA-Pro3 was observed by atomic force microscope (AFM) on highly oriented pyrolytic graphite (HOPG), and the information on the helix direction, pitch, and chain arrangement was obtained. Also, the chiral separation properties of these polymer-based chiral stationary phases (CSPs) were investigated using high-performance liquid chromatography (HPLC). The poly(phenylacetylene)s showed enhanced enantioseparation properties toward various racemates depending on the longer chain length of the L-proline oligopeptide in the pendants and the positive synergy between the helical backbone and helical side chains. Particularly, PPA-Pro9 showed comparable or even superior enantioseparation properties for racemates 2 and 9 to four commercial columns (Daicel Chiralpak or Chiralcel AD, AS, OD, and OT), indicating that these poly(phenylacetylene)-based CSPs have potential practical values. This work presented here provides inspiration for the further development of CSPs based on a new paradigm.

A convenient functionalization strategy of polyimide covalent organic frameworks for uranium-containing wastewater treatment and uranium recovery.

The purpose of this research was to design and synthesize an adsorbent based on polyimide covalent organic frameworks (PICOFs) for uranium-containing wastewater treatment and uranium recovery. A modified solvothermal method was innovatively proposed to synthesize PICOFs with high specific surface area (1998.5 m2 g-1) and regular pore structure. Additionally, a convenient functionalization strategy of PICOFs was designed through polydopamine (PDA) and a well-dispersed polymer (MPC-co-AO) containing multiple functional groups, forming stable composite (PMCA-TPPICOFs) in which the hydrogen bonding and cation-π interactions between PDA and MPC-co-AO played a key role. The obtained PMCA-TPPICOFs as an adsorbent exhibited strong selectivity for uranyl ions (maximum adsorption capacity was 538 mg g-1). In simulated wastewater with low uranium concentrations, the removal rate reached 98.3%, and the concentration of treated simulated wastewater met discharge standards. Moreover, PMCA-TPPICOFs was suitable for fixed-bed column adsorption because of its favorable structure. According to the research about adsorption mechanism, the adsorption primarily relied on electrostatic interaction and complexation. In summary, PMCA-TPPICOFs exhibited good potential for uranium-containing wastewater treatment, expanding the application of PICOFs. And the proposed functionalization strategy and modified solvothermal method may promote research in the fields of material functionalization and COFs synthesis. ENVIRONMENTAL IMPLICATION: Uranium is a raw material for nuclear energy applications, which is toxic and radioactive. If uranium is discharged with wastewater, it would not only pose a threat to the environmental protection and life safety, but also cause the loss of precious nuclear raw materials. Although adsorption was considered to be an effective way to remove uranium, many of the developed adsorbents were difficult to apply due to the harsh wastewater environment and complex preparation processes. This study reported a novel adsorbent and a new functionalization strategy, which was expected to solve the problem of uranium recovery in wastewater.

High glucose-upregulated PD-L1 expression through RAS signaling-driven downregulation of PTRH1 leads to suppression of T cell cytotoxic function in tumor environment.

BACKGROUND: Nearly 80% of patients with pancreatic cancer suffer from glucose intolerance or diabetes. Pancreatic cancer complicated by diabetes has a more immunosuppressive tumor microenvironment (TME) and is associated with a worse prognosis. The relationship between glucose metabolism and programmed cell death-Ligand 1 (PD-L1) is close and complex. It is important to explore the regulation of high glucose on PD-L1 expression in pancreatic cancer and its effect on infiltrating immune effectors in the tumor microenvironment. METHODS: Diabetic murine models (C57BL/6) were used to reveal different immune landscape in euglycemic and hyperglycemic pancreatic tumor microenvironment. Bioinformatics, WB, iRIP [Improved RNA Binding Protein (RBP) Immunoprecipitation]-seq were used to confirm the potential regulating role of peptidyl-tRNA hydrolase 1 homolog (PTRH1) on the stability of the PD-L1 mRNA. Postoperative specimens were used to identify the expression of PD-L1 and PTRH1 in pancreatic cancer. Co-culturing T cells with pancreatic cancer cells to explore the immunosuppressive effect of pancreatic tumor cells. RESULTS: Our results revealed that a high dose of glucose enhanced the stability of the PD-L1 mRNA in pancreatic tumor cells by downregulating PTRH1 through RAS signaling pathway activation following epidermal growth factor receptor (EGFR) stimulation. PTRH1 overexpression significantly suppressed PD-L1 expression in pancreatic cells and improved the proportion and cytotoxic function of CD8+ T cells in the pancreatic TME of diabetic mice. CONCLUSIONS: PTRH1, an RBP, plays a key role in the regulation of PD-L1 by high glucose and is closely related to anti-tumor immunity in the pancreatic TME.

METTL3 promotes pancreatic cancer proliferation and stemness by increasing stability of ID2 mRNA in a m6A-dependent manner.

In eukaryotes, N6-methyladenosine (m6A) is the most prevalent epigenetic alteration. Methyltransferase-like 3 (METTL3) is a key player in the control of m6A, although its function in pancreatic cancer is incompletely understood. In this study, we examined the role that METTL3 plays in pancreatic cancer cell proliferation and stemness. We discovered that in pancreatic cancer cells, METTL3-mediated m6A alterations regulate ID2 as a downstream target. The stability of ID2 mRNA was decreased and m6A modification was effectively eliminated by METTL3 knockdown in pancreatic cancer cells. We also demonstrate that m6a-YTHDF2 is necessary for the METTL3-mediated stabilization of ID2 mRNA. Additionally, we show that ID2 controls the stemness molecules NANOG and SOX2 via the PI3K-AKT pathway to support pancreatic cancer growth and stemness maintenance. Our data suggest that METTL3 may post-transcriptionally upregulate ID2 expression in an m6A-YTHDF2-dependent manner to further promote the stabilization of ID2 mRNA, which may be a new target for pancreatic cancer treatment.

Adjustable extracellular matrix rigidity tumor model for studying stiffness dependent pancreatic ductal adenocarcinomas progression and tumor immunosuppression.

Pancreatic ductal adenocarcinomas (PDAC) is one of the stiffest malignancies with strong solid stresses. Increasing stiffness could alter cellular behavior and trigger internal signaling pathways and is strongly associated with a poor prognosis in PDAC. So far, there has been no report on of an experimental model that can rapidly construct and stably maintain a stiffness gradient dimension in both vitro and in vivo. In this study, a gelatin methacryloyl (GelMA)-based hydrogel was designed for in vitro and in vivo PDAC experiments. The GelMA-based hydrogel has porous, adjustable mechanical properties and excellent in vitro and in vivo biocompatibility. The GelMA-based in vitro 3D culture method can effectively form a gradient and stable extracellular matrix stiffness, affecting cell morphology, cytoskeleton remodeling, and malignant biological behaviors such as proliferation and metastasis. This model is suitable for in vivo studies with long-term maintenance of matrix stiffness and no significant toxicity. High matrix stiffness can significantly promote PDAC progression and tumor immunosuppression. This novel adaptive extracellular matrix rigidity tumor model is an excellent candidate for further development as an in vitro and in vivo biomechanical study model of PDAC or other tumors with strong solid stresses.

Hyperglycemia Enhances Immunosuppression and Aerobic Glycolysis of Pancreatic Cancer Through Upregulating Bmi1-UPF1-HK2 Pathway.

BACKGROUND & AIMS: Accumulating evidence strongly suggests that hyperglycemia promotes the progression of pancreatic cancer (PC). Approximately 80% of patients with PC are intolerant to hyperglycemic conditions. In this study, we define the role of Bmi1, a stemness-related oncogene, in controlling the Warburg effect, and immune suppression under hyperglycemia conditions. METHODS: The diabetes mellitus model was established by intraperitoneal injection of streptozotocin. The role of the hyperglycemia-Bmi1-HK2 axis in glycolysis-related immunosuppression was examined in both orthotopic and xenograft in vivo models. Evaluation of immune infiltrates was carried out by flow cytometry. Human PC cell lines, SW1990, BxPC-3, and CFPAC-1, were used for mechanistic in vitro studies. RESULTS: Through bioinformatics analysis, we found that hyperglycemia was strongly related to aerobic glycolysis, immunosuppression, and cancer cell stemness. High glucose condition in the tumor microenvironment promotes immune suppression by upregulating glycolysis in PC cells, which can be rescued via knockdown Bmi1 expression or after 2-deoxy-D-glucose treatment. Through gain-/loss-of-function assessments, we found that Bmi1 upregulated the expression of UPF1, which enhanced the stability of HK2 mRNA and thereby increased the expression of HK2. The role of the hyperglycemia-Bmi-HK2 pathway in the inhibition of antitumor immunity was further verified via the immune-competent and immunodeficient mice model. We also demonstrated that hyperglycemia promotes the expression of Bmi1 by elevating the intracellular acetyl-CoA levels and histone H4 acetylation levels. CONCLUSIONS: Our results suggest that the previously unreported Bmi1-UPF1-HK2 pathway contributes to PC progression and immunosuppression, which may bring in new targets for developing effective therapies to treat patients with PC.

A novel staging system and clinical predictive nomogram for more accurate staging and prognosis of malignant pancreatic intraductal papillary mucinous neoplasms: a population-based study.

BACKGROUND: The current guidelines of the American Joint Committee on Cancer (AJCC) for the staging of exocrine pancreatic tumors seem inapplicable to malignant pancreatic intraductal papillary mucinous neoplasms (IPMN). Therefore, we aimed to improve the accuracy of clinical staging and prognosis for malignant IPMN by modifiing current AJCC system. METHODS: We extracted data of 2001 patients with malignant IPMN from the Surveillance, Epidemiology, and End Results database between 2000 and 2016. Of these, 1401 patients were assigned to the primary cohort and 600 patients to the validation cohort. RESULTS: In Kaplan-Meier analysis of the primary cohort, the current AJCC guidelines were unable to distinguish between certain tumor substages (IA and IB in the 7th, IB and IIA in the 8th). The modified system that we regrouped based on the median overall survival and hazard ratios, was superior in tumor stage classifications. Age > 70 years, tumors located in the body or tail, high-grade differentiated tumors, surgery, chemotherapy, and tumor, lymph node, and metastasis (TNM) stage were identified as independent predictive factors for overall survival. Compared to that of TNM-based systems, the concordance index of the clinical predictive nomogram significantly improved (0.819; 95% confidence interval, 0.805-0.833), with excellent area under the receiver operating characteristic curves (1-, 3-, and 5-year: 0.881, 0.889, and 0.879, respectively). The calibration curves also showed good agreement between prediction and actual observation. The analysis of treatment modalities revealed that surgery resulted in better survival for all resectable malignant IPMN. The analysis of chemotherapy data reveals its potential in improving the prognosis of treatment for patients with locally advanced or distant metastases. CONCLUSIONS: Our modified staging system improves the distinction of tumor stages. The nomogram was a more accurate and clinically reliable tool for prognosis prediction of patients with malignant IPMN.

Third-Stage Dispersal Juveniles of Bursaphelenchus xylophilus Can Resist Low-Temperature Stress by Entering Cryptobiosis.

Nematodes can enter cryptobiosis by dehydration as an adaptation to low-temperature environments and recover from cryptobiosis by rehydration after environmental improvement. In this work, the survival of Bursaphelenchusxylophilus third-stage dispersal juveniles was studied in response to low-temperature treatment. The average survival rates were 1.7% after -80 °C treatment for 30 d and 82.2% after -20 °C treatment for 30 d. The changes of water content and inorganic salt ions that occur in pine trees during winter gradually alter the osmotic pressure in the liquid environment to dehydrate B. xylophilus juveniles, resulting in improved survival after low-temperature treatment. The survival rate at -20 °C improved to 92.1% when the juveniles entered cryptobiosis by osmotic regulation. The results of this study demonstrate that B. xylophilus third-stage dispersal juveniles can resist low-temperature stress through cryptobiosis, providing the theoretical basis for the identification of areas potentially vulnerable to B. xylophilus in the mid-temperature and cold temperature zones of China.

Construction of gel-like swollen-layer on Polyacrylonitrile Surface and Its Swelling Behavior and Uranium Adsorption Properties.

In this study, a hyperbranched chelated hydrophilic swollen-layer was constructed on the surface of polyacrylonitrile (PAN) fiber with amino trimethylene phosphoric acid (ATMP) as a terminal group, which applied as an adsorbent for seawater uranium U(VI) extraction. This shows that U(VI) enter the gel-like swollen-layer to form a more complex body structure. The molecular chain conformational extension in the swollen-layer reduces the resistance of the uranyl ion to enter the swollen-layer, which is conducive to the adsorption behavior. The adsorption performance on the U(VI) by the adsorption experiment were found to be consistent with the Langmuir isotherm adsorption model and the pseudo-second-order kinetics, indicating that the adsorption of U(VI) by this material is uniform single-layer chemical adsorption. Ion competition experiments and cyclic adsorption experiments verify the practical application potential of the materials. In the dynamic simulation of seawater adsorption experiments, the adsorption capacity of the adsorbent reached 7.4 mg/g. Studies on the adsorption mechanism have found that a large number of hydroxyl groups in the swollen-layer and ATMP as an end machine have a chelation effect on U(VI).

Synthesis and characterization of paclitaxel-imprinted microparticles for controlled release of an anticancer drug.

In this study, novel molecularly imprinted polymer (MIP) microparticles containing methacryl polyhedral oligomeric silsesquioxane (M-POSS) were synthesized through the RAFT precipitation polymerization (RAFTPP) method using paclitaxel (PTX) as the templates. During the course of this investigation, methacrylic acid (MAA) was used as the functional monomer, while ethylene glycol dimethacrylate (EGDMA) was utilized as a cross-linker. The effects of different molar ratios of M-POSS on the microparticles were characterized. The obtained MIP microparticles were confirmed by FT-IR and TGA-DSC. The results of SEM showed regular spherical-shaped MIP microparticles with diameters around 170-490 nm. The PTX loading quantity was closely correlated with the content of M-POSS, and the MIP microparticles showed a satisfactory affinity to PTX with high drug loading (17.1%) and encapsulation efficiency (85.5%). Moreover, these MIP microparticles were sensitive to pH, and consequently the release rates of PTX at pH 5 were much faster than those at pH 7 due to the acid cleavage of the hydrogen bonds. In addition, the results from release experiments of the MIP microparticles showed a very slow and controlled release of PTX, which heralded promising potential as a carrier for PTX delivery in cancer therapy.

Maternal linuron exposure alters testicular development in male offspring rats at the whole genome level.

Linuron is a widely used herbicide; its toxicity on the male reproductive system has been recognized. The current study was designed to explore the molecular mechanism underlying linuron-induced reproductive toxicity. Pregnant rats received daily oral gavage of linuron at the dose of 120mg/kg/d from gestation day (GD)12 to GD17. Tissues from male offspring rats were collected for pathological examination and microarray gene expression profiling. Changes in gene expression were further verified by quantitative real-time RT-PCR. Data showed that linuron-exposed offspring rats had a decreased sperm count (88% of controls) and disrupted acrosome formation. There were evident damages in seminiferous tubules and abnormal morphology in mesenchymal cells in samples from linuron-exposed animals. Microarray analysis indicated that the expressions of testosterone synthesis-associated genes, i.e., Star, P450scc, 3ß-Hsd, Abp, Cox7a2, Pcna, p450c17and17ß-Hsd were significantly altered by linuron exposure, along with other genes involving in cell proliferation and apoptosis, such as c-myc, S6K, Apaf1, and TSC1. These data indicate that linuron upon entering male offspring body can directly or indirectly interact with the androgen production and function; linuron-induced alteration in genes encoding testosterone synthesis is likely a major factor in linuron-induced male reproductive toxicity.

Transformation of dissolved organic matters in swine, cow and chicken manures during composting.

The changes of dissolved organic matters (DOMs) extracted from swine, cow and chicken manures were assessed by Fourier transform infrared, ultraviolet, gel permeation chromatography (GPC), excitation-emission-matrix fluorescence (EEM-FL), Biolog Eco and (1)H NMR during 60-day composting. Pumice was adopted to eliminate the disturbing of common organic bulking agents. The results showed chicken manure had the highest DOC, DTN (dissolved total nitrogen) and lowest DOC/DTN among the three manures; cow manure had the highest volatile solids, lowest DTN, slowest DOMs hydrolysis rate and the fastest bio-stabilization rate. (1)H NMR showed the decrease rates of OC band and saturated carbon chain were distinctly faster than that of olefinic and aromatic structures. The molecular size distribution of DOMs in the three manures was in the range of 1-10 kDa detected by GPC. Microbial carbon utilization capacity decreased in cow manure with composting time, but the contrast was observed in the chicken and swine manures.

µ-4,4'-Diazenediyldiphthalato-κO:O-bis-[penta-aqua-manganese(II)] tetra-hydrate.

The dinuclear complex in the title compound, [Mn(2)(C(16)H(6)N(2)O(8))(H(2)O)(10)]·4H(2)O, lies on an inversion center. Two delocalized carboxyl-ate groups are each connected in a monodentate fashion to two similar penta-aqua-manganese units, whereas the other two localized carboxyl-ate groups are uncoordinated. The metal ion has octa-hedral coordination, with the O atom of a carboxyl-ate group and three coordinated water mol-ecules forming the equatorial plane [Mn-O(COO) = 2.143 (4) Å] and two water mol-ecules occupying the axial positions. The architecture is further consolidated by extensive hydrogen bonds for which coordinated water mol-ecules serve as donors or acceptors.