Cetuximab combined with chemotherapy for simultaneous esophageal squamous cell carcinoma and colon adenocarcinoma: A case report.

BACKGROUND: Multiple primary carcinomas (MPCs) are defined as two or more independent primary cancers that occur simultaneously or sequentially in the same individual. Synchronous MPCs are rarer than solitary cancers or metachronous MPCs. Accurate diagnoses of synchronous MPCs and the choice of treatment are critical for successful outcomes in these cases. CASE SUMMARY: A 64-year-old patient presented with dysphagia, without obvious cause. A diagnosis of synchronous esophageal squamous cell carcinoma and colon adenocarcinoma with liver metastasis was confirmed based on examination and laboratory results. After multi-disciplinary consultations, combination chemotherapy (a 3-wk cycle with oxaliplatin 212 mg administered on day 1 and capecitabine 1.5 g twice daily on days 1-14) and esophageal cancer radiotherapy were initiated. Based on the results of genetic testing, we switched to a regimen of leucovorin + fluorouracil + oxaliplatin and cetuximab regimen for 8 cycles. Subsequently, capecitabine and bevacizumab were administered until the most recent follow-up, at which the tumor remained stable. CONCLUSION: Successful cetuximab chemotherapy treatment provides a reference for the non-operative and homogeneous treatment of different pathological types of synchronous MCPs.

Discovery of toxoflavin, a potent IRE1α inhibitor acting through structure-dependent oxidative inhibition.

Inositol-requiring enzyme 1α (IRE1α) is the most conserved endoplasmic reticulum (ER) stress sensor with two catalytic domains, kinase and RNase, in its cytosolic portion. IRE1α inhibitors have been used to improve existing clinical treatments against various cancers. In this study we identified toxoflavin (TXF) as a new-type potent small molecule IRE1α inhibitor. We used luciferase reporter systems to screen compounds that inhibited the IRE1α-XBP1s signaling pathway. As a result, TXF was found to be the most potent IRE1α RNase inhibitor with an IC50 value of 0.226 µM. Its inhibitory potencies on IRE1α kinase and RNase were confirmed in a series of cellular and in vitro biochemical assays. Kinetic analysis showed that TXF caused time- and reducing reagent-dependent irreversible inhibition on IRE1α, implying that ROS might participate in the inhibition process. ROS scavengers decreased the inhibition of IRE1α by TXF, confirming that ROS mediated the inhibition process. Mass spectrometry analysis revealed that the thiol groups of four conserved cysteine residues (CYS-605, CYS-630, CYS-715 and CYS-951) in IRE1α were oxidized to sulfonic groups by ROS. In molecular docking experiments we affirmed the binding of TXF with IRE1α, and predicted its binding site, suggesting that the structure of TXF itself participates in the inhibition of IRE1α. Interestingly, CYS-951 was just near the docked site. In addition, the RNase IC50 and ROS production in vitro induced by TXF and its derivatives were negative correlated (r = -0.872). In conclusion, this study discovers a new type of IRE1α inhibitor that targets a predicted new alternative site located in the junction between RNase domain and kinase domain, and oxidizes conserved cysteine residues of IRE1α active sites to inhibit IRE1α. TXF could be used as a small molecule tool to study IRE1α's role in ER stress.

Novel yellow solid-state fluorescent-emitting carbon dots with high quantum yield for white light-emitting diodes.

Fluorescence quenching of carbon dots (CDs) occurs in their aggregated state ascribed to direct π-π interactions or excessive resonance energy transfer (RET). Thus, CDs have been severely restricted for applications requiring phosphors that emit in the solid state, such as the fabrication of white light-emitting diodes (WLEDs). In this report, novel CDs with bright solid-state fluorescence (SSF) were synthesized by simple microwave-assisted synthesis method, using 1,4,7,10-tetraazacyclododecane (cyclen) and citric acid as precursors. Under 365 nm UV light, these CDs emit bright yellow SSF, indicating they successfully overcome the aggregation-induced fluorescence quenching (ACQ) effect. When the excitation wavelength (λex) is fixed at 450 nm, the emission peak of the CDs is centered at 546 nm with the Commission Internationale de l'Eclairage chromaticity (CIE) coordinates of (0.43, 0.55), which means that they can be combined with a blue-emitting chip in order to fabricate WLEDs. More importantly, the absolute quantum yield (QY) of these CDs powder reached 48% at λex of 450 nm, which was much higher than many previously reported SSF-emitting CDs and indicating their high light conversion ability in solid-state. Thanks to the excellent optical property of these CDs powder, they were successfully used in the preparation of high-performance WLEDs. This study not only enriches SSF-emitting CD-based nanomaterials with good prospects for application, but also provides valuable reference for subsequent research on the synthesis of solid-state fluorescent CDs.

Algibacter marinivivus sp. nov., isolated from the surface of a marine red alga.

A Gram-stain-negative, non-flagellated bacterium, designated ZY111T, was isolated from the surface of a marine red alga, which was collected from the coast in Weihai, Shandong Province, PR China. Strain ZY111T exhibited growth at 4-37 °C (optimum, 25-28 °C) in the presence of 0-8.0 % (w/v) NaCl (optimum, 2.0-4.0% NaCl) and at pH 6.5-9.5 (optimum, pH 7.0-8.0). The 16S rRNA gene sequence analysis revealed that strain ZY111T belonged to the genus Algibacter, with Algibacter amylolyticus DSM 29199T as its closest relative (97.7 % similarity). The averagenucleotide identity value of strain ZY111T with A. amylolyticus DSM 29199T was 79.03 %. The digitalDNA-DNA hybridization value of strain ZY111T with A. amylolyticus DSM 29199T was 22.40 %. The dominant fatty acids were iso-C15 : 0, iso-C15 : 1 G, iso-C15 : 0 3-OH and iso-C17 : 0 3-OH. The sole respiratory quinone was determined to be menaquinone-6. The polar lipid profile of strain ZY111T consisted of phosphatidylethanolamine, two unidentified aminolipids and three unidentified lipids. The G+C content was 31.9 mol%. The phenotypic, chemotaxonomic and phylogenetic data clearly showed that strain ZY111T represents a novel species of the genus Algibacter, for which the name Algibacter marinivivus sp. nov. is proposed. The type strain is ZY111T (=KCTC 62373T=MCCC 1H00295T).

Biocompatible Photoluminescent Silk Fibers with Stability and Durability.

Exploring photoluminescent silk fibers, possessing biocompatibility as well as stable and durable fluorescent properties, is a requirement for the development of novel photoluminescent biomaterials. Herein, we fabricate photoluminescent silk fibers, TPCA@SF, via modifying an organic fluorescent molecule (5-oxo-3,5-dihydro-2H-thiazolo [3,2-a] pyridine-7-carboxylic acid, TPCA) onto silk fibers, along with using quaternary ammonium salt didodecyldimethylammonium bromide (DDAB) as a color-fixing agent. The hydrogen bonds and electrostatic association among silk fibers, TPCA and DDAB, ensure the stable modification. The facile and green fabrication process is achieved in water under mild conditions without using any toxic substances. The TPCA@SF manifests the combining features of high quantum yield, fluorescence water-fastness, antiphotobleaching, good mechanical property, and biocompatibility. The strategy holds great potential for exploring various biocompatible photoluminescent substances with stability and durability.

pH-sensitive charge-conversional and NIR responsive bubble-generating liposomal system for synergetic thermo-chemotherapy.

A charge-conversional and NIR responsive rapid release liposomal system (PSD/DOX/Cypate-BTSL) was developed to enhance therapeutic efficacy of cancer therapy. The cationic liposomes containing Cypate, doxorubicin (DOX) and NH4HCO3 were shielded by pH-sensitive poly(methacryloyl sulfadimethoxine) (PSD) through electrostatic interaction at pH 7.4. At the tumor site (pH 6.5), PSD was deshielded and the liposomes displayed pH-sensitive charge reversal capability. The DOX released from PSD/DOX/Cypate-BTSL with irradiation was markedly higher than the other groups, indicating NIR irradiation and NH4HCO3 had a significant effect on the drug release. After irradiation, the hyperthermia induced by Cypate could produce CO2 bubbles quickly on account of the decomposition of NH4HCO3, achieving the rapid drug release. In 4T1 cells, PSD/DOX/Cypate-BTSL improved cellular uptake and cytotoxicity with irradiation at pH 6.5. In vivo results implied that the liposomes with irradiation could efficiently enhance the tumor accumulation and antitumor efficacy, and reduce systemic side effects of DOX. In conclusion, PSD/DOX/Cypate-BTSL is a promising candidate as a carrier for synergistic effects of PTT and chemotherapy.

NIR responsive liposomal system for rapid release of drugs in cancer therapy.

To design a rapid release liposomal system for cancer therapy, a NIR responsive bubble-generating thermosensitive liposome (BTSL) system combined with photothermal agent (Cypate), doxorubicin (DOX), and NH4HCO3 was developed. Cypate/DOX-BTSL exhibited a good aqueous stability, photostability, and photothermal effect. In vitro release suggested that the amounts of DOX released from BTSL were obviously higher than that of (NH4)2SO4 liposomes at 42°C. After NIR irradiation, the hyperthermic temperature induced by Cypate led to the decomposition of NH4HCO3 and the generation of a large number of CO2 bubbles, triggering a rapid release of drugs. Confocal laser scanning microscope and acridine orange staining indicated that Cypate/DOX-BTSL upon irradiation could facilitate to disrupt the lysosomal membranes and realize endolysosomal escape into cytosol, improving the intracellular uptake of DOX clearly. MTT and trypan blue staining implied that the cell damage of Cypate/DOX-BTSL with NIR irradiation was more severe than that in the groups without irradiation. In vivo results indicated that Cypate/DOX-BTSL with irradiation could dramatically increase the accumulation of DOX in tumor, inhibit tumor growth, and reduce systemic side effects of DOX. These data demonstrated that Cypate/DOX-BTSL has the potential to be used as a NIR responsive liposomal system for a rapid release of drugs in thermochemotherapy.

Sequential delivery of chlorhexidine acetate and bFGF from PLGA-glycol chitosan core-shell microspheres.

Wound treatment should meet the challenge both of preventing infection and promoting wound healing. To design a sequential delivery system for wound healing, PLGA-glycol chitosan (GC) core-shell microspheres containing chlorhexidine acetate (CHA) at the GC shell and bFGF in the core of PLGA microspheres were fabricated using emulsion-solvent evaporation method. SEM showed that the microspheres were all spherical in shape with a smooth surface. The average size of PLGA-GC microspheres increased due to the GC coating on the surface. The results of release profiles and fluorescence images indicated that PLGA-GC microspheres had an ability to deliver drugs in sequence. The CHA was rapidly released, whereas the proteins presented a sustained release. The release behavior could be modulated by changing the GC amount. Antibacterial assay and cell proliferation tests suggested that the released CHA and bFGF retained their antimicrobial activity and bioactivity during preparation. The microspheres exhibited non-cytotoxicity against 3T3 cells and had a good biocompatibility. These results demonstrated that PLGA-GC core-shell microspheres could be a promising controlled release system of delivering drugs and proteins in sequence for wound healing.

Surface modification of PLGA nanoparticles with biotinylated chitosan for the sustained in vitro release and the enhanced cytotoxicity of epirubicin.

In this study, poly(d,l-lactide-co-glycolide) nanoparticles (PLGA NPs) with biotinylated chitosan (Bio-CS)-surface modification were prepared to be usded as a tumor-targeted and prolonged delivery system for anticancer drugs. Epirubicin (EPB), as a model drug, was encapsulated into Bio-CS surface modified PLGA (Bio-CS-PLGA) NPs with a drug encapsulation efficiency of 84.1 ± 3.4%. EPB-loaded Bio-CS-PLGA NPs were spherical shaped, and had a larger size and higher positive zeta potential compared to the unmodfied EPB-loaded PLGA NPs. The in vitro drug releases showed that EPB-loaded Bio-CS-PLGA NPs exhibited relatively constant drug release kinetics during the first 48 h and the drug burst release significantly decreased in comparison to the unmodified PLGA NPs. The results of MTS assays showed that Bio-CS-PLGA NPs markedly increased the cytotoxicity of EPB, compared to both the unmodified PLGA NPs and the CS-PLGA NPs. The uptakes of NPs in human breast cancer MCF-7 cells were evaluated by the flow cytometry and the confocal microscope. The results revealed that Bio-CS-PLGA NPs exhibited a greater extent of cellular uptake than the unmodified PLGA NPs and CS-PLGA NPs. Moreover, the cellular uptake of Bio-CS-PLGA NPs was evidently inhibited by the endocytic inhibitors and the receptor ligand, indicating that biotin receptor-mediated endocytosis was perhaps involved in the cell entry of Bio-CS-PLGA NPs. In MCF-7 tumor-bearing nude mice, EPB-loaded Bio-CS-PLGA NPs were efficiently accumulated in the tumors. In summary, Bio-CS-PLGA NPs displayed great potential for application as the carriers of anticancer drugs.

Fish collagen-based scaffold containing PLGA microspheres for controlled growth factor delivery in skin tissue engineering.

To design a scaffold controlled release system for skin tissue engineering, fish collagen/chitosan/chondroitin sulfate scaffolds were fabricated by freeze-drying and incorporated with bFGF-loaded PLGA microspheres (MPs). SEM showed that the scaffolds exhibited an interconnected porous structure, and the spherical MPs were uniformly distributed into the scaffolds. The higher swelling and degradation rate of scaffolds/MPs could lead to a higher diffusion rate of MPs from the scaffolds, causing an increase in the protein release. The release rate of proteins could be adjusted by the size of MPs and the ratio of collagen to chitosan of scaffolds. Circular dichroism spectroscopy and MTT of bFGF after release indicated that the released bFGF retained its structural integrity and bioactivity during preparation. Cell proliferation and in vivo evaluation results suggested that the scaffolds/MPs had a good biocompatibility and an ability to promote fibroblast cell proliferation and skin tissue regeneration. These results demonstrated that this scaffold/MP controlled release system has the potential for skin tissue engineering.

Synthesis of Surface-Enhanced Raman Scattering-Active Gold Nanoflowers by 5-Hydroxytryptophan in Acidic Solution.

We demonstrated a one-step synthesis of gold nanoflowers by 5-hydroxytryptophan (5-HTP) in acidic solutions at room temperature. The synthesized nanoflowers are 400 nm in dimension consisting of a solid core and many small petal-like nanoparticles (30-50 nm) in various directions. The shape, size and surface morphology of the gold nanostructure could be tuned by controlling the molar ratio of 5-HTP to HAuCl4 and HCl concentration. Examination of the nanoparticles at different reaction stages with transmission electron microscopy (TEM) reveals the shape evolutionary process of the nanoflowers to get a better understanding on their possible formation mechanism. Additionally, these gold nanoflowers exhibit a high SERS performance because of textured surface which is expected to provide many active sites.

Antitumor Effects and Related Mechanisms of Penicitrinine A, a Novel Alkaloid with a Unique Spiro Skeleton from the Marine Fungus Penicillium citrinum.

Penicitrinine A, a novel alkaloid with a unique spiro skeleton, was isolated from a marine-derived fungus Penicillium citrinum. In this study, the isolation, structure and biosynthetic pathway elucidation of the new compound were described. This new compound showed anti-proliferative activity on multiple tumor types. Among them, the human malignant melanoma cell A-375 was confirmed to be the most sensitive. Morphologic evaluation, apoptosis rate analysis, Western blot and real-time quantitative PCR (RT-qPCR) results showed penicitrinine A could significantly induce A-375 cell apoptosis by decreasing the expression of Bcl-2 and increasing the expression of Bax. Moreover, we investigated the anti-metastatic effects of penicitrinine A in A-375 cells by wound healing assay, trans-well assay, Western blot and RT-qPCR. The results showed penicitrinine A significantly suppressed metastatic activity of A-375 cells by regulating the expression of MMP-9 and its specific inhibitor TIMP-1. These findings suggested that penicitrinine A might serve as a potential antitumor agent, which could inhibit the proliferation and metastasis of tumor cells.

Collagen/chitosan film containing biotinylated glycol chitosan nanoparticles for localized drug delivery.

The objective of this study was to design a drug delivery system consisting of biotinylated cholesterol-modified glycol chitosan (Bio-CHGC) nanoparticles and fish collagen/chitosan (Col/Ch) film for localized chemotherapy. Bio-CHGC was synthesized, and then its self-assembled nanoparticles were prepared by probe sonication. Doxorubicin (DOX)-loaded Bio-CHGC (DBC) nanoparticles prepared by dialysis had spherical shape, and their sizes were in the range of 330-397 nm. Col/Ch/DBC nanoparticle films were fabricated by freeze-drying. SEM showed that the DBC nanoparticles were uniformly distributed into the films, and the films retained their structural integrity. A higher degradation and swelling rate of the drug films led to a higher diffusion rate of the nanoparticles from the films, resulting in an increase in the drug release from nanoparticles. The release of DOX from the films or Bio-CHGC nanoparticles was sensitive to the pH value of the release medium. In addition, the DOX release ratio of the drug films was lower than that of the nanoparticles alone, suggesting that the drug films had a double-sustained effect on the drug release. MTT assay implied that the DBC nanoparticle film showed a higher inhibitory ratio than the film containing nanoparticles without biotin, indicating that biotin moieties in the nanoparticles played an important role in exerting a cytotoxic effect. These data demonstrate that Col/Ch/DBC nanoparticle film has the potential to be used as a localized delivery system for hydrophobic antitumor drugs.

[Study of collagen mimetic peptide's triple-helix structure and its thermostability by circular dichroism].

In the present study, the authors explore the triple-helix conformation and thermal stability of collagen mimetic peptides (CMPs) as a function of peptide sequence and/or chain length by circular dichroism(CD). Five CMPs were designed and synthetized varying the number of POG triplets or incorporating an integrin alpha2beta1 binding motif Gly-Phe-Hyp-Gly-Glu-Arg (GFOGER). CD spectroscopy from 260 to 190 nm was recorded to confirm the existence of triple-helix conformation at room temperature, while thermal melting and thermal annealing of triple-helix (thermal unfolding and refolding of triple-helix, respectively) was characterized by monitoring ellipticity at 225 nm as a function of temperature. The results demonstrated that all the CMPs adopted triple-helix conformation, and the thermal stability of the CMPs was enhanced with increasing the number of POG triplets. In contrast to natural collagen, the thermal denaturation processes of CMPs were reversible, i. e. the triple-helix unfolded upon heating while refolded upon cooling. Meanwhile, the phenomenon of "hysteresis" was observed by comparing melting and thermal curves. These findings add new insights to the mechanisms of collagen and CMPs assembly, as well as provide an alternative approach to the fabrication of artificial collagen-likes biomaterials.

Speradines F-H, three new oxindole alkaloids from the marine-derived fungus Aspergillus oryzae.

A rare hexacyclic oxindole alkaloid, speradine F (1), together with two novel tetracyclic oxindole alkaloids, speradines G (2) and H (3), were isolated from the marine-derived fungus Aspergillus oryzae. Their structures were determined by spectroscopic methods and X-ray diffraction analysis. This study is the first report on cyclopiazonic acid (CPA)-type alkaloids with a hexacyclic skeleton.

The marine fungal metabolite, dicitrinone B, induces A375 cell apoptosis through the ROS-related caspase pathway.

Dicitrinone B, a rare carbon-bridged citrinin dimer, was isolated from the marine-derived fungus, Penicillium citrinum. It was reported to have antitumor effects on tumor cells previously; however, the details of the mechanism remain unclear. In this study, we found that dicitrinone B inhibited the proliferation of multiple tumor types. Among them, the human malignant melanoma cell, A375, was confirmed to be the most sensitive. Morphologic evaluation, cell cycle arrest and apoptosis rate analysis results showed that dicitrinone B significantly induced A375 cell apoptosis. Subsequent observation of reactive oxygen species (ROS) accumulation and mitochondrial membrane potential (MMP) reduction revealed that the apoptosis induced by dicitrinone B may be triggered by over-producing ROS. Further studies indicated that the apoptosis was associated with both intrinsic and extrinsic apoptosis pathways under the regulation of Bcl-2 family proteins. Caspase-9, caspase-8 and caspase-3 were activated during the process, leading to PARP cleavage. The pan-caspase inhibitor, Z-VAD-FMK, could reverse dicitrinone B-induced apoptosis, suggesting that it is a caspase-dependent pathway. Our data for the first time showed that dicitrinone B inhibits the proliferation of tumor cells by inducing cell apoptosis. Moreover, compared with the first-line chemotherapy drug, 5-fluorouracil (5-Fu), dicitrinone B showed much more potent anticancer efficacy, suggesting that it might serve as a potential antitumor agent.

Purification and characterization of a gelatinolytic matrix metalloproteinase from the skeletal muscle of grass carp (Ctenopharyngodon idellus).

A gelatinolytic matrix metalloproteinase (gMMP) from grass carp skeletal muscle was purified by 30-70% ammonium sulphate fractionation and a combination of chromatographic steps including ion exchange on DEAE-Sephacel, gel filtration on Sephacryl S-200, and affinity on gelatin-sepharose. The molecular weight of the proteinase as estimated by SDS-PAGE was 70 kDa under non-reducing conditions. The enzyme revealed high activity from 30 to 50 °C, and the gelatin hydrolysing activity was investigated at a slightly alkaline pH range using gelatin as substrate. Metalloproteinase inhibitor EDTA completely suppressed the gelatinolytic activity, while other proteinase inhibitors did not show any inhibitory effect. Divalent metal ion Ca(2+) was essential for the gelatinolytic activity. Further, peptide mass fingerprinting obtained four fragments with 45 amino acid residues, which were highly identical to MMP-2 from fish species. The gMMP could effectively hydrolyse type I collagen even at 4 °C, suggesting its involvement in the texture softening of fish muscle during the post-mortem stage.

In vitro degradation of three-dimensional chitosan/apatite composite rods prepared via in situ precipitation.

Three-dimensional (3D) compact rods with multilayer structure made from chitosan (CHI) and apatite (Ap) have been prepared. The cytocompatibility assay revealed that the CHI/Ap composite could promote cell proliferation. In vitro degradation behaviors of the rods have been systematically investigated for up to 6 weeks in phosphate buffer saline (PBS) solution at 37°C. The properties of the composite rods were measured by means of weight loss, swelling ratio, and the changes in mechanical properties, etc. The pH of the PBS solution during the first 2 weeks of degradation was also detected. Results showed that the medium of CHI/Ap composite rods exhibited more stable pH change compared with that of CHI rods. Weight loss as well as the changes in mechanical properties happened more often to CHI rods than CHI/Ap rods. The presence of Ap could effectively reduce the degradation rate of the composite rods. All the results suggested that the composite rods could keep the initial shapes and mechanical properties longer than the pure CHI rods.

Fabrication of chitosan/hydroxylapatite composite rods with a layer-by-layer structure for fracture fixation.

A composite rod for fracture fixation using chitosan (CHI)/hydroxylapatite (HA) was prepared by means of in situ precipitation, which had a layer-by-layer structure, good mechanical properties, and cell compatibilities. The CHI/HA composite rods were precipitated from the chitosan solution with calcium and phosphorus precursors, followed by treatment with a tripolyphosphate-trisodium phosphate solution (pH >13) to crosslink the CHI and to hydrolyze the calcium phosphates to nanocrystalline HA. The results of FTIR, XRD, and TEM measurements confirmed that HA had been formed within the CHI matrix. The effects of the CHI/HA ratios (20/0, 20/1, 20/2, 20/4, and 20/5, w/w) on the mechanical properties were investigated. At the CHI/HA ratio of 20/4 (w/w), the bending strength and modulus of the rods were 133 MPa and 6.8 GPa, respectively. Pre-osteoblast MC3T3-E1 cells were cultured in an extract of the CHI/HA rods (20/4, w/w) to study the cell compatibilities of the composite. The observations indicated that the CHI/HA composite could promote the growth of MC3T3-E1 cells better than the composite without HA (p < 0.05). Furthermore, the co-cultivation of the cells and the CHI/HA composite showed that cells fully spread on the surface of the composite with an obvious cytoskeleton organization, which also revealed that the CHI/HA composite had a good biocompatibility.