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BACKGROUND & AIMS: As the first approved medication for metabolic dysfunction-associated steatohepatitis (MASH), thyroid hormone receptor-beta (THR-ß) agonist MGL-3196 (Resmetirom) is highly spotlighted as the liver-directed, bioactive oral drug. However, it was also identified with remarkable heterogeneity of individual clinical efficacy and its interference with gut microbiota in host hepatoenteral circulation was still undocumented. METHODS: We compared MASH attenuation by MGL-3196 and its derivative drug HSK31679 between germ-free (GF) and specific-pathogen free (SPF) mice to evaluate the role of gut microbiota. Then cross-omics analyses of microbial metagenome, metabolome and single-cell RNA-sequencing were applied into the randomized, double-blind, placebo-controlled multiple-ascending-dose (MAD) cohort of HSK31679 treatment (n = 40), to comprehensively investigate the altered gut microbiota metabolism and circulating immune signatures. RESULTS: HSK31679 outperformed MGL-3196 in ameliorating MASH diet-induced steatohepatitis of SPF mice but not GF mice. In the MAD cohort of HSK31679, relative abundance of B. thetaiotaomicron was significantly enriched to impair glucosylceramide synthase (GCS)-catalyzed monoglucosylation of microbial Cer(d18:1/16:0) and Cer(d18:1/24:1). In stark contrast to the non-inferiority MASH resolution between MGL-3196 and HSK31679 for GFBTΔGCS mice, HSK31679 manifested superior steatohepatitis alleviation than MGL-3196 for GFBTWT mice, due to its steric hindrance with R123 and Y401 of gut microbial GCS. For participants with high fecal GCS activity, the administration of 160 mg HSK31679 induced a shift in peripheral compartments towards an immunosuppressive niche, characterized by decreased CD8α+ dendritic cells and MINCLE+ macrophages. CONCLUSIONS: This study provided novel insights into the indispensable gut microbiota for HSK31679 treatment, which revealed microbial GCS may serve as its prognostic biomarker of MASH treatment, as well as the new target for further strategies of microbiota-based MASH therapeutics. IMPACT AND IMPLICATIONS: Remarkable heterogeneity of individual clinical efficacy of THR-ß agonists and their interferences with microbiome in host hepatoenteral circulation are poorly understood. In our current germ-free mice models and randomized, double-blind multiple-dose cohort study, we identified microbial GCS as the prognostic biomarker of HSK31679 treatment, as well as the new target for further strategies of microbiota-based MASLD therapeutics.
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OBJECTIVES: Colitis is a global disease usually accompanied by intestinal epithelial damage and intestinal inflammation, and an increasing number of studies have found natural products to be highly effective in treating colitis. Anemoside B4 (AB4), an abundant saponin isolated from Pulsatilla chinensis (Bunge), which was found to have strong anti-inflammatory activity. However, the exact molecular mechanisms and direct targets of AB4 in the treatment of colitis remain to be discovered. METHODS: The anti-inflammatory activities of AB4 were verified in LPS-induced cell models and 2, 4, 6-trinitrobenzene sulfonic (TNBS) or dextran sulfate sodium (DSS)-induced colitis mice and rat models. The molecular target of AB4 was identified by affinity chromatography analysis using chemical probes derived from AB4. Experiments including proteomics, molecular docking, biotin pull-down, surface plasmon resonance (SPR), and cellular thermal shift assay (CETSA) were used to confirm the binding of AB4 to its molecular target. Overexpression of pyruvate carboxylase (PC) and PC agonist were used to study the effects of PC on the anti-inflammatory and metabolic regulation of AB4 in vitro and in vivo. RESULTS: AB4 not only significantly inhibited LPS-induced NF-κB activation and increased ROS levels in THP-1 cells, but also suppressed TNBS/DSS-induced colonic inflammation in mice and rats. The molecular target of AB4 was identified as PC, a key enzyme related to fatty acid, amino acid and tricarboxylic acid (TCA) cycle. We next demonstrated that AB4 specifically bound to the His879 site of PC and altered the protein's spatial conformation, thereby affecting the enzymatic activity of PC. LPS activated NF-κB pathway and increased PC activity, which caused metabolic reprogramming, while AB4 reversed this phenomenon by inhibiting the PC activity. In vivo studies showed that diisopropylamine dichloroacetate (DADA), a PC agonist, eliminated the therapeutic effects of AB4 by changing the metabolic rearrangement of intestinal tissues in colitis mice. CONCLUSION: We identified PC as a direct cellular target of AB4 in the modulation of inflammation, especially colitis. Moreover, PC/pyruvate metabolism/NF-κB is crucial for LPS-driven inflammation and oxidative stress. These findings shed more light on the possibilities of PC as a potential new target for treating colitis.
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Colite , Saponinas , Ratos , Camundongos , Animais , Piruvato Carboxilase/metabolismo , NF-kappa B/metabolismo , Lipopolissacarídeos/farmacologia , Simulação de Acoplamento Molecular , Colite/induzido quimicamente , Colite/tratamento farmacológico , Colite/metabolismo , Inflamação/metabolismo , Saponinas/farmacologia , Anti-Inflamatórios/farmacologia , Anti-Inflamatórios/uso terapêutico , Macrófagos/metabolismo , Sulfato de Dextrana/efeitos adversos , Sulfato de Dextrana/metabolismo , Camundongos Endogâmicos C57BL , Modelos Animais de DoençasRESUMO
BACKGROUND: Hepatocellular carcinoma (HCC) has a high incidence and mortality worldwide, which seriously threatens people's physical and mental health. Coagulation is closely related to the occurrence and development of HCC. Whether coagulation-related genes (CRGs) can be used as prognostic markers for HCC remains to be investigated. METHODS: Firstly, we identified differentially expressed coagulation-related genes of HCC and control samples in the datasets GSE54236, GSE102079, TCGA-LIHC, and Genecards database. Then, univariate Cox regression analysis, LASSO regression analysis, and multivariate Cox regression analysis were used to determine the key CRGs and establish the coagulation-related risk score (CRRS) prognostic model in the TCGA-LIHC dataset. The predictive capability of the CRRS model was evaluated by Kaplan-Meier survival analysis and ROC analysis. External validation was performed in the ICGC-LIRI-JP dataset. Besides, combining risk score and age, gender, grade, and stage, a nomogram was constructed to quantify the survival probability. We further analyzed the correlation between risk score and functional enrichment, pathway, and tumor immune microenvironment. RESULTS: We identified 5 key CRGs (FLVCR1, CENPE, LCAT, CYP2C9, and NQO1) and constructed the CRRS prognostic model. The overall survival (OS) of the high-risk group was shorter than that of the low-risk group. The AUC values for 1 -, 3 -, and 5-year OS in the TCGA dataset were 0.769, 0.691, and 0.674, respectively. The Cox analysis showed that CRRS was an independent prognostic factor for HCC. A nomogram established with risk score, age, gender, grade, and stage, has a better prognostic value for HCC patients. In the high-risk group, CD4+T cells memory resting, NK cells activated, and B cells naive were significantly lower. The expression levels of immune checkpoint genes in the high-risk group were generally higher than that in the low-risk group. CONCLUSIONS: The CRRS model has reliable predictive value for the prognosis of HCC patients.
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Carcinoma Hepatocelular , Neoplasias Hepáticas , Humanos , Prognóstico , Carcinoma Hepatocelular/genética , Neoplasias Hepáticas/genética , Nomogramas , Fatores de Risco , Microambiente TumoralRESUMO
In this letter, a sub-pm linewidth, high pulse energy and high beam quality microsecond-pulse 766.699â nm Ti:sapphire laser pumped by a frequency-doubled Nd:YAG laser is demonstrated. At an incident pump energy of 824 mJ, the maximum output energy of 132.5 mJ at 766.699â nm with linewidth of 0.66 pm and a pulse width of 100 µs is achieved at a repetition rate of 5â Hz. To the best of our knowledge, this is the highest pulse energy at 766.699â nm with pulse width of hundred micro-seconds for a Ti:sapphire laser. The beam quality factor M2 is measured to be 1.21. It could be precisely tuned from 766.623 to 766.755â nm with a tuning resolution of 0.8 pm. The wavelength stability is measured to be less than ±0.7 pm over 30 min. The sub-pm linewidth, high pulse energy and high beam quality Ti:sapphire laser at 766.699â nm can be used to create a polychromatic laser guide star together with a home-made 589â nm laser in the mesospheric sodium and potassium layer for the tip-tilt correction resulting in the near-diffraction limited imagery on a large telescope.
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PURPOSE: Posterior reversible encephalopathy syndrome (PRES) displayed various acute neurological symptoms. PRES is a rare presentation of hypercalcemia. Here we present a case with ectopic secretion of parathyroid hormone from neuroendocrine carcinoma of the endometrium presenting as hypercalcemia-related PRES. CASE REPORT: A 67-year-old woman presented with acute generalized tonic-clonic seizure followed by post-ictal confusion and neuropsychiatric behaviors. The diagnosis is PRES. Investigations showed uterine cervical region with multiple liver metastasis complicated with hypercalcemia, elevated intact parathyroid hormone. Further pathology concluded as a poorly differentiated adenocarcinoma of the endometrium with neuroendocrine differentiation and immunoreactive for PTH. The patient's neurologic manifestations had resolved. Serum free calcium level and intact-PTH had declined after first course of definitive chemoradiation. CONCLUSION: Immunostaining of the tumor tissue can be used to estimate the ectopic PTH production within the tumor cells. Early detection and appropriate clinical treatment hold the potential to improve the prognosis of refractory hypercalcemia and hypercalcemia related PRES. Keyword: Posterior reversible encephalopathy syndrome; hypercalcemia; intact-parathyroid hormone; parathyroid hormone-related peptide; neuroendocrine carcinoma of endometrium.
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Carcinoma Neuroendócrino , Hipercalcemia , Síndrome da Leucoencefalopatia Posterior , Idoso , Carcinoma Neuroendócrino/complicações , Endométrio , Feminino , Humanos , Hipercalcemia/etiologia , Hormônio ParatireóideoRESUMO
Carcinosarcomas are biphasic tumors comprising carcinoma and sarcoma components that occur in many tissues but are rarely found in the orbit. A 70-year-old male presented to the ophthalmic clinic with progressive proptosis, having decreased vision in the left eye for 8 months. On examination, severe exophthalmos and lagophthalmos with limited extraocular movement were noted. Orbital computed tomography scans revealed a large, well-defined, heterogeneously enhanced mass in the left retrobulbar orbital cavity. The tumor was completely resected, and the pathological examination revealed a carcinosarcoma. The prognosis was excellent without local recurrence at 48 months postoperatively. Thus, when considering treatment for effective management of such tumors, tumor resection followed by radiotherapy or chemotherapy is highly recommended.
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Carcinossarcoma , Exoftalmia , Idoso , Carcinossarcoma/diagnóstico por imagem , Carcinossarcoma/radioterapia , Carcinossarcoma/cirurgia , Exoftalmia/etiologia , Humanos , Masculino , Órbita/diagnóstico por imagem , Órbita/cirurgia , Prognóstico , Tomografia Computadorizada por Raios XRESUMO
Background Tirapazamine's (TPZ) tolerability after an intra-arterial (IA) injection remains unclear. We investigated TPZ's safety and tolerability in rats by first injecting into the left hepatic artery and then performing a hepatic artery ligation, which recapitulates the transarterial embolization used clinically. Research design and methods: Forty-six rats in five groups were respectively injected with 0, 0.25, 0.50, 1.0, or more than 1.5 mL IA of TPZ (0.7 mg/mL) into the left hepatic artery and then subjected to hepatic artery ligation under laparotomy. Blood samples were collected four times daily up to day 15 after which the rats were euthanized and necropsied. The toxicity profile of IA injection of TPZ followed by hepatic artery ligation was then assessed. Results No significant changes to the rats' body weight and serum total bilirubin were observed. Serum alanine aminotransferase (ALT) levels increased slightly but remained below 100 U/L one day after treatment for most rats. Three rats in Groups 3 and 4 exhibited an over two-fold transient elevation of ALT. All ALT recovered to the baseline at day 14. Liver tissues were collected on day 15 using H&E staining. One rat in Group 3 showed ischemic coagulative necrosis in its liver tissue. Other sporadic pathological changes not related to TPZ doses were observed in Groups 2, 3, 4, and 5. Conclusion TPZ by IA injection followed by embolization is tolerated up to 7 mg/kg. This finding supports the strategy of administering an IA injection of TPZ followed by trans-arterial embolization to the liver.
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Antineoplásicos/toxicidade , Tirapazamina/toxicidade , Alanina Transaminase/sangue , Animais , Bilirrubina/sangue , Feminino , Artéria Hepática/cirurgia , Injeções Intra-Arteriais , Ligadura , Fígado/efeitos dos fármacos , Fígado/patologia , Masculino , Ratos , Hipóxia TumoralRESUMO
Photoresponsive materials are a key part of the age of smart technology that have potential in a broad range of applications. Coordination networks (CNs) are widely used due to their designability and stability. In this work, three novel alkaline earth metal coordination networks (AEM-CNs): [Mg(CMNDI)(H2 O)2 ], [Ca(CMNDI)(H2 O)2 ]â H2 O, and [Sr(CMNDI)(H2 O)(DMF)] with fsl, cds, and scn topology nets were synthetized via N,N'-bis(carboxymethyl)-1,4,5,8-naphthalenediimide (H2 CMNDI); the scn net is not found in the Reticular Chemistry Structure Resource or ToposPro. The reusable and sensitive photochromic properties of the three CNs enable them to be used as secret inks or ultraviolet detectors. In addition, the CNs also exhibited reusable photoluminescent turn-off toward the drug molecules, balsalazide disodium (Bal.) and colchicine (Col.), with good limits of detection of 0.16 and 0.70â µM. To the best of our knowledge, this is the first study of a fluorescence sensor for Bal. Thus, the AEM-CNs provide a design idea for integrated photoresponsive materials that could be further improved in the near future by further study.
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Metais AlcalinoterrososRESUMO
Juniperus indica Bertol. is an herbal plant that belongs to the genus Juniperus, which is commonly used in traditional medicine to refresh the mind and for diuretic use. However, few studies have reported the function of J. indica Bertol. Hence, this study aimed to investigate the anti-tumor and synergistic potential of J. indica Bertol. extract (JIB extract) for melanoma cells. Our results indicated the anti-melanoma activity of JIB extract. JIB extract induced cell cycle arrest at the G0/G1 phase and decreased cyclin and cdk protein expressions. In addition, AKT/mTOR signaling and MAPK signaling were inhibited by JIB extract to suppress melanoma cell growth and proliferation. Additionally, JIB extract induced B16/F10 cell apoptosis via the caspase cascade. According to the JIB extract's anti-melanoma capacity, to assess the synergistic effects of cisplatin and JIB extract. The results demonstrated that JIB extract combined with cisplatin enhanced the inhibition of cell growth, proliferation, and survival through the obstruction of cell cycle progression and AKT/mTOR and MAPK signaling as well as the induction of cell apoptosis. Collectively, our results indicate that JIB extract showed anti-tumor effects and synergized with cisplatin against B16/F10 cells, indicating the possibility of JIB extract to be developed as adjuvant therapy for melanoma.
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Protocolos de Quimioterapia Combinada Antineoplásica/farmacologia , Cisplatino/farmacologia , Juniperus/química , Melanoma/tratamento farmacológico , Extratos Vegetais/farmacologia , Neoplasias Cutâneas/tratamento farmacológico , Animais , Protocolos de Quimioterapia Combinada Antineoplásica/uso terapêutico , Apoptose/efeitos dos fármacos , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Cisplatino/uso terapêutico , Cães , Sinergismo Farmacológico , Pontos de Checagem da Fase G1 do Ciclo Celular/efeitos dos fármacos , Humanos , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Células Madin Darby de Rim Canino , Melanoma/patologia , Camundongos , Extratos Vegetais/uso terapêutico , Proteínas Proto-Oncogênicas c-akt/metabolismo , Transdução de Sinais/efeitos dos fármacos , Serina-Treonina Quinases TOR/metabolismoRESUMO
BACKGROUND: pH-sensitive peptides are a relatively new strategy for conquering the poor endosomal release of cationic polymer-mediated transfection. Modification of antimicrobial peptides by exchanging positively-charged residues with negatively-charged glutamic acid residues (Glu) greatly improves its lytic activity at the endosomal pH, which could improve cationic polymer-mediated transfection. METHODS: In the present study, we investigated the effect of the number of Glu substituted for positively-charged residues on the endosomal escape activity of AR-23 and the ability of mutated AR-23 with respect to enhancing cationic polymer-mediated transfection. Three analogs were synthesized by replacing the positively-charged residues in the AR-23 sequence with Glu one-by-one. RESULTS: The pH-sensitive lysis ability of the peptides, the effect of peptides on the physicochemical characteristics, the intracellular trafficking, the transfection efficiency and the cytotoxicity of the polyplexes were determined. Increased lytic activity of peptides was observed with the increased number of Glu replacement in the AR-23 sequence at acidic pH. The number of Glu substituted for positively-charged residues of AR-23 dramatically affects its lysis ability at neutral pH. Triple-Glu substitution in the AR-23 sequence greatly improved poly(l-lysine)-mediated gene transfection efficiency at the same time as maintaining low cytotoxicity. CONCLUSIONS: The results indicate that replacement of positively-charged residues with sufficient Glu residues may be considered as a method for designing pH-sensitive peptides, which could be applied as potential enhancers for improving cationic polymer-mediated transfection.
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DNA/administração & dosagem , Endossomos/efeitos dos fármacos , Terapia Genética , Hemólise/efeitos dos fármacos , Neoplasias/terapia , Polilisina/química , Proteínas Citotóxicas Formadoras de Poros/farmacologia , Apoptose , Proliferação de Células , Técnicas de Transferência de Genes , Humanos , Concentração de Íons de Hidrogênio , Neoplasias/genética , Neoplasias/patologia , Proteínas Citotóxicas Formadoras de Poros/química , Células Tumorais CultivadasRESUMO
A 100 W level kilohertz repetition-rate microsecond (µs)-pulse all-solid-state sodium beacon laser at 589 nm is demonstrated for the first time, to the best of our knowledge, via combining two independent µs-pulsed lasers. Each beamlet is generated by the sum-frequency mixing of pulsed 1064 and 1319 nm lasers in a lithium triborate (LBO) crystal, which operate at 500 Hz pulse repetition frequency with 61 W $p$p-polarized and 53 W $s$s-polarized output, respectively. An incoherent sequence combining technology of polarized laser beams is employed to add the two beamlets. The average power of the combined beam is up to 107.5 W with a combining efficiency of 94.3%. The combined beam has a 1 kHz repetition rate with ${\sim}{120}\;\unicode{x00B5} {\rm s}$â¼120µs pulse duration and beam quality ${M^2} = {1.41}$M2=1.41. The central wavelength with a linewidth of ${\sim}{0.3}\;{\rm GHz}$â¼0.3GHz is locked to a sodium ${{\rm D}_{2a}}$D2a absorption line. To the best of our knowledge, this is a record-high power operating at kilohertz for µs-pulsed solid-state sodium beacon lasers.
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A void-free bonding technique was demonstrated for a large slab Nd: YAG crystal with a bonding surface dimension of â¼160mm×70mm. By using the novel fluxless oxide layer removal technology, the indium-oxide barrier problem was resolved. With the help of electrochemical-polished indium solder and a plasma-cleaned heat sink, the solderability of the indium was enhanced; in particular, the contact angle of the solder was improved from 51° to 31°. With the largest-bonding-size slab, a single-slab laser created a maximum output power of 7.3 kW under an absorbed pump power of 12.8 kW, corresponding to an optical to optical efficiency of 57% and a slope conversion of 67.8%. By detecting the wavefront of the interferometer before and after bonding, the RMS of wavefront was 0.192λ and 0.434λ (λ=633nm), respectively. To the best of our knowledge, this is the largest void-free bonding size for a laser slab and the highest output power achieved from a single-slab crystal laser oscillator.
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This paper was aimed to observe the effect of anemoside B4(hereinafter referred to as B4) on cisplatin-induced acute kidney injury in mice, and to investigate its possible mechanism in renal protection from inflammation and apoptosis aspects. Mice were divided into normal group, model group, dexamethasone positive group and B4 high, middle and low dose groups(5, 2.5, and 1.25 mg·kg~(-1 )doses). All the other mice groups except normal group were given with tail vein injection of cisplatin(15 mg·kg~(-1)) to induce acute kidney injury models. The drug administration was started on the day of modeling, and lasted for 4 days. After 1 hour of the last injection, orbital blood was collected. After the serum was separated, serum urea nitrogen(BUN), creatinine(Cre), total protein(TP), and albumin(ALB) were tested by using an automatic biochemical analyzer; the changes of kidney pathological morphology were observed by PAS staining; the protein expression levels of inflammatory factors including nucleotide binding oligomerization domain-like receptor(NLRP3), cysteinyl aspartate specific proteinase 1(caspase-1), interleukin-18(IL-18), interleukin-1ß(IL-1ß), tumor necrosis factor(TNF-α), and interleukin-6(IL-6) and apoptosis factors including p53, caspase-3, cleaved-caspase-3, Bcl-2 associated X protein(Bax), and B-cell lymphoma-2(Bcl-2) were analyzed by Western blot. The results showed that B4 significantly reduced the serum BUN and Cre contents, and alleviated pathological changes in renal tissues, such as the shedding and degeneration of renal tubular epithelial cells, tubulin tubule type. B4 significantly down-regulated the protein expressions of p53, Bax, cleaved-caspase-3 in the kidney and up-regulated the expression of Bcl-2/Bax. In model group, however, no significant up-regulation was observed in the protein expression levels of inflammatory cytokines(NLRP3, pro-caspase-1, IL-18, IL-1ß, TNF-α, IL-6). The results suggested that B4 had a certain protective effect on cisplatin-induced acute kidney injury, and could activate p53 signaling pathway related apoptotic factors. B4 renal protective effect was mainly related to the regulation of p53 signaling pathway, while NLRP3 inflammasome and related inflammatory factors had no obvious response in this model.
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Injúria Renal Aguda/tratamento farmacológico , Apoptose , Inflamação , Saponinas/uso terapêutico , Injúria Renal Aguda/induzido quimicamente , Animais , Proteínas Reguladoras de Apoptose , Citocinas , Rim , CamundongosRESUMO
Immediate and delayed hypersensitivity reactions may occur after the first or many exposures to tocilizumab, and they have varying presentations. Here, we describe a Wolf's isotopic response that manifested in a patient as erythema on the same site of a previous healed herpes zoster infection. This phenomenon has rarely been reported in the literature.
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Anticorpos Monoclonais Humanizados/efeitos adversos , Antirreumáticos/efeitos adversos , Toxidermias/etiologia , Eritema/etiologia , Herpes Zoster/complicações , Idoso , Artrite Reumatoide/tratamento farmacológico , Feminino , HumanosRESUMO
Camellia oleifera C. Abel has been widely cultivated in China, and a group of bioactive constituents such as triterpeniod saponin have been isolated from C. oleifera C. Abel. In the current study, a new triterpeniod saponin was isolated from the EtOH extract of the roots of C. oleifera C. Abel, named as oleiferoside W, and the cytotoxic properties of oleiferoside W were evaluated in non-small cell lung cancer A549 cells. At the same time the inducing apoptosis, the depolarization of mitochondrial membrane potential (Δψ), the up-regulation of related pro-apoptotic proteins, such as cleaved-PARP, cleaved-caspase-3, and the down-regulation of anti-apoptotic marker Bcl-2/Bax were measured on oleiferoside W. Furthermore, the function, inducing the generation of reactive oxygen species (ROS) and apoptosis, of oleiferoside W could be reversed by N-acetylcysteine (NAC). In conclusion, our findings showed that oleiferoside W induced apoptosis involving mitochondrial pathway and increasing intracellular ROS production in the A549 cells, suggesting that oleiferoside W may have the possibility to be a useful anticancer agent for therapy in lung cancer.
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Antineoplásicos Fitogênicos/farmacologia , Apoptose/efeitos dos fármacos , Camellia/química , Pontos de Checagem do Ciclo Celular/efeitos dos fármacos , Raízes de Plantas/química , Saponinas/farmacologia , Triterpenos/farmacologia , Células A549 , Antineoplásicos Fitogênicos/administração & dosagem , Antineoplásicos Fitogênicos/química , Relação Dose-Resposta a Droga , Humanos , Estrutura Molecular , Espécies Reativas de Oxigênio , Saponinas/administração & dosagem , Saponinas/química , Triterpenos/administração & dosagem , Triterpenos/químicaRESUMO
Advanced melanoma can metastasize to distal organs from the skin and yield an aggressive disease and poor prognosis even after treatment with chemotherapeutic agents. The compound n-Butylidenephthalide (BP) is isolated from Angelica sinensis, which is used to treat anemia and gynecological dysfunction in traditional Chinese medicine. Studies have indicated that BP can inhibit cancers, including brain, lung, prostate, liver, and colon cancers. However, because BP is a natural hydrophobic compound, it is quickly metabolized by the liver within 24 h, and thus has limited potential for development in cancer therapy. This study investigated the anticancer mechanisms of BP through encapsulation with a novel polycationic liposome containing polyethylenimine (PEI) and polyethylene glycol complex (LPPC) in melanoma cells. The results demonstrated that BP/LPPC had higher cytotoxicity than BP alone and induced cell cycle arrest at the G0/G1 phase in B16/F10 melanoma cells. The BP/LPPC-treated cell indicated an increase in subG1 percentage and TUNEL positive apoptotic morphology through induction of extrinsic and intrinsic apoptosis pathways. The combination of BP and LPPC and clinical drug 5-Fluorouracil had a greater synergistic inhibition effect than did a single drug. Moreover, LPPC encapsulation improved the uptake of BP values through enhancement of cell endocytosis and maintained BP cytotoxicity activity within 24 h. In conclusion, BP/LPPC can inhibit growth of melanoma cells and induce cell arrest and apoptosis, indicating that BP/LPPC has great potential for development of melanoma therapy agents.
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Antineoplásicos/administração & dosagem , Lipossomos , Melanoma , Anidridos Ftálicos/administração & dosagem , Poliaminas , Angelica sinensis/química , Animais , Antineoplásicos/química , Antineoplásicos/isolamento & purificação , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Portadores de Fármacos/química , Humanos , Lipossomos/química , Melanoma/tratamento farmacológico , Melanoma/metabolismo , Camundongos , Anidridos Ftálicos/química , Anidridos Ftálicos/isolamento & purificação , Polieletrólitos , Polietilenoglicóis/química , Polietilenoimina/químicaRESUMO
BACKGROUND: Poor endosomal release is a major barrier of polyplex-mediated gene transfection. Antimicrobial peptides (AMPs) are commonly used to improve polyethylenimine (PEI)-mediated gene transfection by increasing endosomal release. In the present study, we designed novel pH-sensitive peptides that highly enhance transfection efficiency compared to their parent peptides. METHODS: Two analogues of melittin (Mel) and RV-23 (RV) were synthesized by replacing the positively-charged residues in their sequences with glutamic acid residues. The pH-sensitive lysis ability of the peptides, the effect of the peptides on physicochemical characteristics, the intracellular trafficking, the transfection efficiency, and the cytotoxicity of the polyplexes were determined. RESULTS: The acidic peptides showed pH-sensitive lytic activity. The hemolytic activity of acidic peptides at pH 5.0 was higher than that at pH 7.4. The incorporation of acidic peptides did not affect the DNA binding ability of PEI but affected the physicochemical characteristics of the PEI/DNA polyplexes, which may be beneficial for endosomal release and gene transfection. The incorporation of acidic peptides into PEI/DNA polyplexes enhanced the PEI-mediated transfection efficiency corresponding to up to 42-fold higher luciferase activity compared to that of PEI alone. CONCLUSIONS: The results of the present study indicate that replacement of positively-charged residues with glutamic acid residues in the AMP sequence yields pH-sensitive peptides, which enhance the transfection efficiency of PEI/DNA polyplexes in various cell lines.
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Anti-Infecciosos/química , Peptídeos/química , Polietilenoimina/química , Anti-Infecciosos/farmacologia , Linhagem Celular , Linhagem Celular Tumoral , DNA/metabolismo , Endossomos/metabolismo , Técnicas de Transferência de Genes , Células HeLa , Células Hep G2 , Células Endoteliais da Veia Umbilical Humana , Humanos , Concentração de Íons de Hidrogênio , Meliteno/química , Peptídeos/administração & dosagem , Peptídeos/farmacologia , Transfecção/métodosRESUMO
Filler migration is a potential complication following the injection of multiple fillers. With the increasing popularity of multiple filler injections, migrated granulomas should be an essential differential diagnosis for newly growing facial lumps. It is important for all physicians to be aware that complication induced by dermal fillers can occur in locations other than the planned injected sites. We described a case of filler migration to the forehead in a patient addicted to cosmetic fillers. To our knowledge, it has never been published in dermatology literature so far. A detailed history of cosmetic procedures from the patient addicted to filler injections is necessary for accurate diagnosis. Because account of previous cosmetic filler injections is not always reliable, an early skin biopsy with pathological examination is the gold standard for determining whether multiple filler injections have been performed.
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Técnicas Cosméticas/efeitos adversos , Preenchedores Dérmicos/efeitos adversos , Testa , Migração de Corpo Estranho/patologia , Poliésteres/efeitos adversos , Feminino , Humanos , Pessoa de Meia-IdadeRESUMO
Cluster of differentiation 44 (CD44) and octamer-binding transcription factor 3/4 (Oct3/4) are important factors influencing cancer stem cell (CSC) development, but their clinical applications on pancreatic cancer are still unknown. Here, we tested the hypothesis that expression of CD44 and Oct3/4 correlates with the clinicopathological parameters of pancreatic ductal adenocarcinomas (PDACs). Firstly, data on the mRNA expression levels in PDACs and normal pancreatic tissues were collected from Gene Expression Omnibus (GEO) repository datasets. Immunohistochemical analyses of CD44 and Oct3/4 were next performed in tissue microarrays of 80 surgical specimens derived from a Chinese population, which included 9 normal pancreatic ducts and 71 PDACs, amongst which 12 were well differentiated, 47 moderately differentiated, and 12 poorly differentiated. From the GEO results, mRNA expression levels of both CD44 and Oct3/4 were higher in PDACs than in normal pancreatic tissues. In addition, immunostaining scores of these biomarkers were higher in most PDACs than in non-neoplastic pancreatic ducts. The intensity of CD44 and Oct3/4 staining in normal pancreatic tissues was weak and limited to small areas. Although CD44 and Oct3/4 overexpression in PDACs tended to be associated with advanced histologic grades of PDACs, the correlation of CD44 and Oct3/4 expression with the American Joint Committee on Cancer (AJCC) pathological stage was not statistically significant. In conclusion, CD44 and Oct3/4 overexpression may imply malignant transformation of pancreatic ducts and could help pathologists make a more accurate diagnosis and decision on clear surgical margins.
Assuntos
Adenocarcinoma/metabolismo , Biomarcadores Tumorais/metabolismo , Receptores de Hialuronatos/metabolismo , Fator 3 de Transcrição de Octâmero/metabolismo , Ductos Pancreáticos/metabolismo , Neoplasias Pancreáticas/metabolismo , Adenocarcinoma/patologia , Feminino , Perfilação da Expressão Gênica/métodos , Humanos , Masculino , Ductos Pancreáticos/patologia , Neoplasias Pancreáticas/patologia , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , Células Tumorais CultivadasRESUMO
OBJECTIVE: Alcohol dehydrogenase (ADH) and aldehyde dehydrogenase (ALDH) are principal enzymes responsible for the metabolism of ethanol. East Asian populations are unique in that they carry both a prevalent ADH1B*2 and a dominant-negative ALDH2*2 allele. A systematic investigation of ethanol-metabolizing activities in normal livers correlated with the corresponding functional allelic variations and protein contents of the relevant isozymes in respective enzyme families has been lacking. MATERIALS AND METHODS: To obtain a reasonable sample size encompassing all possible genetic allelotypes of the ADH1B and ALDH2, 141 surgical liver specimens from adult Han Chinese were studied. Expression patterns and activities of ADH and ALDH were determined with stratification of the genetic phenotypes. Absolute protein contents as well as cellular localization of the activity and protein of ADH/ALDH isozymes were also investigated. RESULTS: The activities of ADH1B*1/*2 and ADH1B*2/*2 allelic phenotypes were 5-6-fold those of the ADH1B*1/*1, suggesting that ADH1B*2 allele-encoded subunits are dominant over expression of hepatic ADH activity. The activities of the ALDH2-active phenotype were 90% higher than those of the ALDH2-inactive phenotype. Sex and age did not significantly influence the hepatic ADH and ALDH activities with specified genetic phenotypes. The isozyme protein contents were as follows in decreasing order: ADH1, ADH2, ALDH1A1, ALDH2, and ADH3. Both ADH1, but not ADH2/3, and ALDH1A1/2 showed a preferential expression in perivenular hepatocytes. CONCLUSION: Functional correlations of ADH1B*2 and ALDH2*2 variant alleles in the liver provide a biochemical genetic basis suggesting their contribution toward variability in ethanol metabolism as well as susceptibility to alcoholism and alcohol-related diseases in East Asians.