ABSTRACT
Antimicrobial resistance poses a significant global challenge, demanding innovative approaches, such as the CRISPR-Cas-mediated resistance plasmid or gene-curing system, to effectively combat this urgent crisis. To enable successful curing of antimicrobial genes or plasmids through CRISPR-Cas technology, the development of an efficient broad-host-range delivery system is paramount. In this study, we have successfully designed and constructed a novel functional gene delivery plasmid, pQ-mini, utilizing the backbone of a broad-host-range Inc.Q plasmid. Moreover, we have integrated the CRISPR-Cas12f system into the pQ-mini plasmid to enable gene-curing in broad-host of bacteria. Our findings demonstrate that pQ-mini facilitates the highly efficient transfer of genetic elements to diverse bacteria, particularly in various species in the order of Enterobacterales, exhibiting a broader host range and superior conjugation efficiency compared to the commonly used pMB1-like plasmid. Notably, pQ-mini effectively delivers the CRISPR-Cas12f system to antimicrobial-resistant strains, resulting in remarkable curing efficiencies for plasmid-borne mcr-1 or blaKPC genes that are comparable to those achieved by the previously reported pCasCure system. In conclusion, our study successfully establishes and optimizes pQ-mini as a broad-host-range functional gene delivery vector. Furthermore, in combination with the CRISPR-Cas system, pQ-mini demonstrates its potential for broad-host delivery, highlighting its promising role as a novel antimicrobial tool against the growing threat of antimicrobial resistance.
Subject(s)
Anti-Bacterial Agents , CRISPR-Cas Systems , Gram-Negative Bacteria , Plasmids , CRISPR-Cas Systems/genetics , Plasmids/genetics , Gram-Negative Bacteria/genetics , Gram-Negative Bacteria/drug effects , Anti-Bacterial Agents/pharmacology , Drug Resistance, Bacterial/genetics , Gene Transfer Techniques , Gene Editing/methodsABSTRACT
Mycophenolate mofetil (MMF) is commonly utilized for the treatment of neuromyelitis optica spectrum disorders (NMOSD). However, a subset of patients experience significant gastrointestinal (GI) adverse effects following MMF administration. The present study aims to elucidate the underlying mechanisms of MMF-induced GI toxicity in NMOSD. Utilizing a vancomycin-treated mouse model, we compiled a comprehensive data set to investigate the microbiome and metabolome in the GI tract to elucidate the mechanisms of MMF GI toxicity. Furthermore, we enrolled 17 female NMOSD patients receiving MMF, who were stratified into non-diarrhea NMOSD and diarrhea NMOSD (DNM) groups, in addition to 12 healthy controls. The gut microbiota of stool samples was analyzed using 16S rRNA gene sequencing. Vancomycin administration prevented weight loss and tissue injury caused by MMF, affecting colon metabolomes and microbiomes. Bacterial ß-glucuronidase from Bacteroidetes and Firmicutes was linked to intestinal tissue damage. The DNM group showed higher alpha diversity and increased levels of Firmicutes and Proteobacteria. The ß-glucuronidase produced by Firmicutes may be important in causing gastrointestinal side effects from MMF in NMOSD treatment, providing useful information for future research on MMF. IMPORTANCE: Neuromyelitis optica spectrum disorder (NMOSD) patients frequently endure severe consequences like paralysis and blindness. Mycophenolate mofetil (MMF) effectively addresses these issues, but its usage is hindered by gastrointestinal (GI) complications. Through uncovering the intricate interplay among MMF, gut microbiota, and metabolic pathways, this study identifies specific gut bacteria responsible for metabolizing MMF into a potentially harmful form, thus contributing to GI side effects. These findings not only deepen our comprehension of MMF toxicity but also propose potential strategies, such as inhibiting these bacteria, to mitigate these adverse effects. This insight holds broader implications for minimizing complications in NMOSD patients undergoing MMF therapy.
Subject(s)
Disease Models, Animal , Gastrointestinal Microbiome , Mycophenolic Acid , Neuromyelitis Optica , Mycophenolic Acid/adverse effects , Mycophenolic Acid/therapeutic use , Neuromyelitis Optica/drug therapy , Neuromyelitis Optica/microbiology , Humans , Animals , Mice , Gastrointestinal Microbiome/drug effects , Female , Adult , Middle Aged , Vancomycin/adverse effects , RNA, Ribosomal, 16S/genetics , Gastrointestinal Tract/drug effects , Gastrointestinal Tract/microbiology , Diarrhea/chemically induced , Diarrhea/microbiology , Male , Gastrointestinal Diseases/chemically induced , Feces/microbiology , Bacteria/drug effects , Bacteria/genetics , Bacteria/classificationABSTRACT
Mycophenolate mofetil (MMF) is a viable therapeutic option against various immune disorders as a chemotherapeutic agent. Nevertheless, its application has been undermined by the gastrotoxic metabolites (mycophenolic acid glucuronide, MPAG) produced by microbiome-associated ß-glucuronidase (ßGUS). Therefore, controlling microbiota-produced ßGUS underlines the potential strategy to improve MMF efficacy by overcoming the dosage limitation. In this study, the octyl gallate (OG) was identified with promising inhibitory activity on hydrolysis of PNPG in our high throughput screening based on a chemical collection of approximately 2000 natural products. Furthermore, OG was also found to inhibit a broad spectrum of BGUSs, including mini-Loop1, Loop 2, mini-Loop 2, and mini-Loop1,2. The further in vivo experiments demonstrated that administration of 20â¯mg/kg OG resulted in predominant reduction in the activity of BGUSs while displayed no impact on the overall fecal microbiome in mice. Furthermore, in the MMF-induced colitis model, the administration of OG at a dosage of 20â¯mg/kg effectively mitigated the gastrointestinal toxicity, and systematically reverted the colitis phenotypes. These findings indicate that the OG holds promising clinical potential for the prevention of MMF-induced gastrointestinal toxicity by inhibition of BGUSs and could be developed as a combinatorial therapy with MFF for better clinical outcomes.
Subject(s)
Colitis , Gallic Acid/analogs & derivatives , Gastrointestinal Microbiome , Mice , Animals , Mycophenolic Acid/pharmacology , Mycophenolic Acid/therapeutic use , Immunosuppressive Agents/therapeutic use , Glucuronidase/metabolism , Bacteria/metabolism , Colitis/drug therapyABSTRACT
BACKGROUND: Neuromyelitis optica spectrum disorder (NMOSD) is a recurring inflammatory demyelinating disease that is commonly observed in Asian countries like China. Prior investigations have shown that mycophenolate mofetil (MMF) with better biocompatibility compared to azathioprine (AZA), and can prevent relapses of NMOSD, but the efficacy was controversially reported in different NMOSD cases. We aimed to explore the factors that weaken efficacy of MMF in NMOSD. METHODS: A total of 34 NMOSD patients treated with MMF were prospectively enrolled and grouped according to the therapeutic efficacy as effective group (EG, n = 23) versus less-effective group (LEG, n = 11). The purine metabolites were profiled in serum samples and gut microbiota was analyzed using 16S rRNA sequencing with stool samples from the same patients. RESULTS: Purine salvage pathway (PSP) metabolites (inosine, hypoxanthine, xanthine, guanine and uric acid) in the serum of NMOSD patients were elevated in the LEG compared to EG (p < 0.05). Additionally, the richness and microbial diversity of gut microbiota was found to be similar between EG and LEG patients. However, LEG patients had increased presence of Clostridium and Synergistes but decreased abundance of the Coprococcus genus. CONCLUSIONS: The PSP metabolites and composition of the gut microbiota were changed between patients with or without optimal clinical response after MMF treatment. This may help us to understand the pharmacodynamics of MMF in NMOSD.
Subject(s)
Gastrointestinal Microbiome , Neuromyelitis Optica , Humans , Mycophenolic Acid/therapeutic use , Neuromyelitis Optica/drug therapy , RNA, Ribosomal, 16S , Treatment Outcome , Azathioprine/therapeutic use , RecurrenceABSTRACT
BACKGROUND: Stem cell-based therapies have demonstrated great potential in several clinical trials. However, safety data on stem cell application remain inadequate. This study evaluated the toxicity of human umbilical cord mesenchymal stem cells (hUC-MSCs) in NOD/Shi-scid/IL-2 Rγnull (NOG) mice. RESEARCH DESIGN AND METHODS: Mice were administered hUC-MSCs intravenously at doses of 3.5 × 106 cells/kg and 3.5 × 107 cells/kg. Toxicity was assessed by clinical observation, behavioral evaluation, pathology, organ weight, and histopathology. We determined the distribution of hUC-MSCs using a validated qPCR method and colonization using immunohistochemistry. RESULTS: No significant abnormal effects on clinical responses, body weight, or food intake were observed in the mice, except for two in the high-dose group that died during the last administration. Mouse activity in the high-dose group decreased 6 h after the first administration. Terminal examination revealed dose-dependent changes in hematology. The mice in the high-dose group displayed pulmonary artery wall plaques and mild alveolar wall microthrombi. hUC-MSCs colonized primarily the lung tissues and were largely distributed there 24 h after the final administration. CONCLUSIONS: The no observed adverse effect level for intravenous administration of hUC-MSCs in NOG mice over a period of 3 w was 3.5 × 106 cells/kg.
Subject(s)
Mesenchymal Stem Cells , Umbilical Cord , Humans , Mice , Animals , Injections, Intravenous , Mice, Inbred NOD , Lung , Mesenchymal Stem Cells/physiologyABSTRACT
Ganoderma lucidum spore powder is a traditional Chinese medicine with a variety of health benefits. Sporoderm-removed Ganoderma lucidum spores (RGLS) can be more effectively absorbed and utilized by the body. Due to the extensive clinical application and lack of long-term (>30 days) safety evaluation of RGLS, it is necessary to evaluate its repeated dose toxicity during a longer administration period. Here, we conducted a 26-week repeated dose toxicity test of RGLS in SpragueâDawley (SD) rats. The male and female rats were orally administered RGLS at doses of 0, 0.4, 1.2, and 4.0 g/kg once daily for a period of 26 weeks. The safety profile of RGLS was assessed through in vivo observations of survival, body weight, and food consumption; hematological, biochemical, and urine analyses; immunotoxicity assays; and histopathological examinations. The results showed that no significant systemic toxicity was observed following 26 weeks of repeated RGLS administration. Our data showed a no-observed adverse effect level (NOAEL) of 4.0 g/kg, which is approximately 20 times higher than the human equivalent dose. Our results support that RGLS can be considered a safe medicinal or food product that can be added to a healthy diet.
Subject(s)
Ganoderma , Reishi , Humans , Rats , Male , Female , Animals , Spores, Fungal , Rats, Sprague-Dawley , Medicine, Chinese Traditional , No-Observed-Adverse-Effect LevelABSTRACT
The fall armyworm, Spodoptera frugiperda (Noctuidae: Lepidoptera), is a wide-reaching notorious insect pest of important cereal crops, which has developed resistance to multiple classes of insecticides. It invaded the Sichuan Province of China in 2019. In this study, we performed resistance monitoring of insecticides for 11 field-collected populations from Sichuan, and all the populations were susceptible to emamectin benzoate and chlorpyrifos. The variations in resistance level to indoxacarb (resistance ratio (RR), 9.23-45.53-fold), spinetoram (RR, 4.32-18.05-fold), and chlorantraniliprole (RR, 2.02-10.39-fold) were observed among these populations. To investigate the resistance mechanism of chlorantraniliprole, synergism tests were performed and showed that piperonyl butoxide had a slight synergistic effect on chlorantraniliprole for the QJ-20 population (1.43-fold) in moderate resistance (RR, 10.39-fold) compared with the treatment group without synergist. Furthermore, the expression scanning for resistance-related genes showed that five P450 genes (CYP6AE43, CYP321A8, CYP305A1, CYP49A1, and CYP306A1) and the ryanodine receptor gene (Ryr, chlorantraniliprole target) were overexpressed in the QJ-20 population. These results indicated that the fall armyworm in Sichuan has exhibited diverse susceptibilities to several classes of insecticides, and the overexpression of Ryr and several P450 genes may contribute to the development of resistance in S. frugiperda to chlorantraniliprole.
ABSTRACT
[This corrects the article DOI: 10.3389/fphys.2023.1155455.].
ABSTRACT
Introduction: Spodoptera frugiperda is an important nomadic agricultural pest with a diverse host range and resistance against several insecticides. The current study investigated the life history traits of two strains of the field-collected population against chlorantraniliprole using an age-stage two-sex life table. Method: For this, we established the chlorantraniliprole-susceptible (Crp-SUS G12), and chlorantraniliprole-reduced susceptible (Crp-RES G12) strains derived from the sixth generation of the QJ-20 population having a resistance ratio (RR) of 10.39-fold, compared with the reported susceptible population. Results: The results showed that the chlorantraniliprole-reduced susceptible strain attained a 4.0-fold RR, while the chlorantraniliprole-susceptible strain attained an RR of 0.85-fold, having overlapped fiducial limits (FLs) with the referred susceptible baseline. Meanwhile, the present study revealed that the development time of the susceptible strain was significantly longer than that of the reduced susceptible strain. Similarly, the mean longevity, adult pre-oviposition period (APOP), and total pre-oviposition period (TPOP) of the female chlorantraniliprole-susceptible strain were considerably longer than those of the female chlorantraniliprole-reduced susceptible strain. Contrarily, the population parameters, including the intrinsic rate of increase (r), finite rate of increase (λ), and net reproductive rate (R), of the chlorantraniliprole-susceptible strain were considerably lower than those of the chlorantraniliprole-reduced susceptible strain, while the mean generation time (T) of the chlorantraniliprole-susceptible strain was substantially longer than the chlorantraniliprole-reduced susceptible strain. The age-stage characteristic survival rate (s xj ) and age-stage characteristic life expectancy (e xj ) of the chlorantraniliprole-susceptible strain were longer than those of the chlorantraniliprole-reduced susceptible strain, but the age-stage-specific reproductive value (v xj ) of the chlorantraniliprole-susceptible strain was shorter than that of the chlorantraniliprole-reduced susceptible strain. Moreover, the contents of vitellogenin (Vg) and VgR in the chlorantraniliprole-reduced susceptible strain were higher than those in the chlorantraniliprole-susceptible strain. Discussion: These findings showed that reducing susceptibility to chlorantraniliprole promoted population growth in S. frugiperda. Therefore, this study could provide conceptual support for the integrated pest management (IPM) approach to control S. frugiperda in the field.
ABSTRACT
Proinflammatory signaling is a hallmark feature of human cancer, including in myeloproliferative neoplasms (MPNs), most notably myelofibrosis (MF). Dysregulated inflammatory signaling contributes to fibrotic progression in MF; however, the individual cytokine mediators elicited by malignant MPN cells to promote collagen-producing fibrosis and disease evolution are yet to be fully elucidated. Previously, we identified a critical role for combined constitutive JAK/STAT and aberrant NF-κB proinflammatory signaling in MF development. Using single-cell transcriptional and cytokine-secretion studies of primary cells from patients with MF and the human MPLW515L (hMPLW515L) murine model of MF, we extend our previous work and delineate the role of CXCL8/CXCR2 signaling in MF pathogenesis and bone marrow fibrosis progression. Hematopoietic stem/progenitor cells from patients with MF are enriched for a CXCL8/CXCR2 gene signature and display enhanced proliferation and fitness in response to an exogenous CXCL8 ligand in vitro. Genetic deletion of Cxcr2 in the hMPLW515L-adoptive transfer model abrogates fibrosis and extends overall survival, and pharmacologic inhibition of the CXCR1/2 pathway improves hematologic parameters, attenuates bone marrow fibrosis, and synergizes with JAK inhibitor therapy. Our mechanistic insights provide a rationale for therapeutic targeting of the CXCL8/CXCR2 pathway among patients with MF.
Subject(s)
Myeloproliferative Disorders , Neoplasms , Primary Myelofibrosis , Humans , Mice , Animals , Primary Myelofibrosis/pathology , Myeloproliferative Disorders/genetics , Signal Transduction , Neoplasms/complications , Cytokines/metabolism , Janus Kinase 2/genetics , Janus Kinase 2/metabolismABSTRACT
The co-occurrence of plasmid-mediated multidrug resistance and hypervirulence in epidemic carbapenem-resistant Klebsiella pneumoniae has emerged as a global public health issue. In this study, an ST23 carbapenem-resistant hypervirulent K. pneumoniae (CR-HvKP) strain VH1-2 was identified from cucumber in China and harbored a novel hybrid plasmid pVH1-2-VIR. The plasmid pVH1-2-VIR carrying both virulence and multidrug-resistance (MDR) genes was likely generated through the recombination of a virulence plasmid and an IncFIIK conjugative MDR plasmid in clinical ST23 18622 isolated from a sputum sample. The plasmid pVH1-2-VIR exhibited the capacity for transfer to the clinical ST11 carbapenem-resistant K. pneumoniae (CRKP) strain via conjugation assay. Acquisition of pVH1-2-VIR plasmid directly converted a CRKP into CR-HvKP strain characterized by hypermucoviscosity, heightened virulence for Galleria mellonella larvae, and increased colonization ability in the mouse intestine. The emergence of such a hybrid plasmid may expedite the spread of CR-HvKP strains, posing a significant risk to human health.
ABSTRACT
Current therapy for myelofibrosis (MF) results in a limited prolongation of patient survival. In order to improve treatment outcomes, we developed a strategy to effectively deplete MF hematopoietic stem/progenitor cells (HSPCs). In the present study, an imipridone, ONC201, was combined with RG7112, an antagonist of MDM2, a p53 negative regulator, to activate downstream events of the p53 and TNF-related apoptosis-inducing ligand (TRAIL)/death receptor (DR) pathways. As compared to treatment with the individual drugs, the combination of ONC201 and RG7112 promoted greater degrees of apoptosis of MF CD34+ cells through activation of both p53-dependent and -independent pathways. Importantly, treatment with ONC201-RG7112 not only decreased the number of JAK2V617F+ and calreticulin mutated colonies assayed from MF CD34+ cells, but allowed for the persistence or appearance of JAK2 wild type colonies. Treatment with ONC201 combined with RG7112 could be a potentially effective strategy for treating MF patients.
Subject(s)
Antineoplastic Agents/pharmacology , Hematopoietic Stem Cells/drug effects , Imidazoles/pharmacology , Imidazolines/pharmacology , Primary Myelofibrosis/drug therapy , Pyridines/pharmacology , Pyrimidines/pharmacology , Tumor Suppressor Protein p53/metabolism , Cells, Cultured , Drug Delivery Systems , Hematopoietic Stem Cells/metabolism , Humans , Primary Myelofibrosis/metabolism , Signal Transduction/drug effectsABSTRACT
A series of vegetable oil-based waterborne polyurethane composites were prepared through construction of novel semi-interpenetrating polymers network using carboxymethyl chitosan (CA) as the secondary polymer phase. The effects of CA contents on storage stability, and particle size distribution of the composite dispersions and thermal stability, mechanical properties and surface wettability of composite films were investigated and discussed. The results showed that the composite dispersions displayed excellent storage stability and the biomass contents of resulting films were high up to 80 %. A significant increase in crosslinking density and glass transition temperature of the composite films were observed as the CA contents increased, which was attributed to the increasing hard segment of films and strong hydrogen bonding interaction between polyurethanes and CA. This work provided a simple method to tailor the performance of environmentally friendly vegetable oil-based waterborne polyurethane, which could find application in the field of coatings, adhesives, ink and so on.
Subject(s)
Castor Oil/chemistry , Chitosan/analogs & derivatives , Polyurethanes/chemistry , Water/chemistry , Chitosan/chemistry , Drug Stability , Humans , Hydrogen Bonding , Materials Testing , Particle Size , Phase Transition , Tensile Strength , Transition Temperature , WettabilityABSTRACT
RATIONALE: Pituitary stalk interruption syndrome (PSIS) is a congenital pituitary anatomical defect. It is characterized by the triad of thin or interrupted pituitary stalk, absent or ectopic posterior lobe, and hypoplastic or aplastic anterior lobe. Moreover, this condition is considered rare. PATIENT CONCERNS: A 23-year-old male patient presented with a history of short stature and hypogonadism. Laboratory assessment revealed low thyroxine, cortisol, and adrenocorticotropic hormone levels, which are consistent with adrenal insufficiency without hypoglycemia. The insulin-induced hypoglycemia tolerance test finding indicated growth hormone (GH) deficiency. Moreover, magnetic resonance imaging revealed an interrupted pituitary stalk, ectopic posterior pituitary, and hypoplastic anterior pituitary. This triad of symptoms was indicative of PSIS. DIAGNOSIS: INTERVENTIONS:: The patient was deficient in adrenaline, thyroxine, gonadal steroid, and GH. Thus, glucocorticoid replacement therapy was initiated, followed by euthyrox, androgen, and human chorionic gonadotropin treatment. Calcium tablets, calcitriol, and alendronate sodium were used for the management of osteoporosis. The patient was 164âcm tall, and his bone age was approximately 15 years old. However, owing to a poor economic condition, the family did not proceed with GH therapy. OUTCOMES: The patient did not present with adrenal or hypothyroidism crisis after receiving poly-hormonal replacement therapy. His secondary sexual characteristics began to develop. However, owing to a short treatment window period, the patient could not receive the required treatment. Hence, whether the patient would have a normal fertility function needs to be confirmed. LESSONS: PSIS is a rare disease with various clinical characteristics. During the neonatal period and infancy, the signs and symptoms of PSIS are often not evident. Therefore, diagnosis is delayed. The early detection of hormone deficiency and treatment initiation can affect both the quality of life and the prognosis of patients with PSIS. Thus, the diagnosis and treatment of this disease must be improved to help patients achieve a better quality of life and to prevent reproductive health problems.
Subject(s)
Pituitary Gland/abnormalities , Adrenocorticotropic Hormone/deficiency , Glucose Tolerance Test , Growth Disorders/etiology , Humans , Hydrocortisone/deficiency , Hypogonadism/etiology , Magnetic Resonance Imaging , Male , Pituitary Gland/diagnostic imaging , Pituitary Gland, Anterior/abnormalities , Pituitary Gland, Posterior/abnormalities , Pituitary-Adrenal System , Thyroxine/deficiency , Young AdultABSTRACT
Sanghuangporus vaninii, called "Sanghuang," is orally used for health care, tumor, and inflammation treatment in Asia. However, the safety of S. vaninii has not been evaluated. The major compounds analysis showed that aqueous extracts of S. vaninii fruiting body were rich in polysaccharides, nucleotides, and polyphenols. Then, the aqueous was given orally to Sprague-Dawley rats for toxical test. In acute toxicity study, the maximum tolerated dose was 21 g/kg. In 17-week repeated dose toxicity experiment, all rats had no abnormal reaction among control group, low dose group (0.15 g/kg) and middle dose group (1.00 g/kg). At high dose group (6.00 g/kg), the feces began to darken on 16th day (D16), and turned to drug stained stool on D21, all rats recovered on the 3rd day (D92) of recovery period. During the whole experiment, there were no animal death and no treatment-related changes in any of the parameters under the all doses. These results indicated the No-Observed Adverse Effect Level of aqueous extract of S. vaninii fruiting body was 1.0 g/kg.
ABSTRACT
OBJECTIVE: To analyze the detection rate of thyroid nodules and its influencing factors among individuals undergoing routine health check-up in Guangdong Province. METHODS: We analyzed the results of thyroid ultrasonography of 309 576 individuals receiving routine health check-up in a general hospital in Guangdong Province over the past 10 years. The data were compared between the individuals with and without thyroid nodules using two independent samples t test, MannWhitney U test and Chi-square test. Binary logistic regression was used to analyze the relationship between different factors and thyroid nodules, and the odds ratio (OR) and 95% confidence interval (95% CI) were calculated. RESULTS: Based on the structured and standardized data, the total detection rate of thyroid nodules was 9.68% among the individuals, 7.71% in male and 13.51% in female subjects, showing a significant gender difference (χ2=2677.08, P < 0.001). In the overall population, the detection rates of thyroid nodules gradually increased with age, but in each age group, the detection rate remained significantly higher in female subjects than in male subjects (P < 0.001). The subjects with thyroid nodules showed a significant higher percentage of male gender, an older age, a greater BMI, and significantly higher SBP, DBP, FBG, TC, LDL-C, HDL-C, TG as well as higher detection rates of fatty liver, hyperlipidemia, hyperglycemia, and metabolic syndrome than those without thyroid nodules (all P < 0.001). Binary logistic regression analysis showed that thyroid nodules were significantly associated with gender (OR=0.455, 95%CI: 0.443-0.468), age (45-59 years: OR=1.660, 95%CI: 1.613-1.710; ≥ 60 years: OR=3.329, 95%CI: 3.202- 3.462), BMI (underweight: OR=0.808, 95%CI: 0.755-0.864; overweight: OR=1.074, 95%CI: 1.038-1.112; obesity: OR=1.281, 95%CI: 1.221-1.343), hyperlipidemia (OR=1.053, 95%CI:1.022-1.085), high blood glucose (OR=1.177, 95%CI: 1.105-1.252), and metabolic syndrome (OR=1.111, 95%CI: 1.071-1.152). CONCLUSIONS: The detection rate of thyroid nodule is much higher in female than in male individuals in Guangdong Province and is significantly associated with gender, age, BMI and metabolic related diseases. Male gender and underweight might be protective factors for thyroid nodules, while an advanced age, overweight/obesity, hyperlipidemia, high blood glucose and metabolic syndrome are associated with an increased risk of thyroid nodules. Thyroid ultrasound examination is recommended in routine health check-up for early detection and treatment of thyroid diseases.
Subject(s)
Thyroid Nodule , Adolescent , Adult , Female , Humans , Male , Middle Aged , Overweight , Risk Factors , Surveys and Questionnaires , Young AdultABSTRACT
A limited number of drugs are available to treat patients with polycythemia vera (PV) and essential thrombocythemia (ET). We attempted to identify alternative agents that may target abnormalities within malignant hematopoietic stem (HSCs) and progenitor cells (HPCs). Previously, MDM2 protein levels were shown to be upregulated in PV/ET CD34+ cells, and exposure to a nutlin, an MDM2 antagonist, induced activation of the TP53 pathway and selective depletion of PV HPCs/HSCs. This anticlonal activity was mediated by upregulation of p53 and potentiated by the addition of interferon-α2a (IFN-α2a). Therefore, we performed an investigator-initiated phase 1 trial of the oral MDM2 antagonist idasanutlin (RG7388; Roche) in patients with high-risk PV/ET for whom at least 1 prior therapy had failed. Patients not attaining at least a partial response by European LeukemiaNet criteria after 6 cycles were then allowed to receive combination therapy with low-dose pegylated IFN-α2a. Thirteen patients with JAK2 V617F+ PV/ET were enrolled, and 12 (PV, n = 11; ET, n = 1) were treated with idasanutlin at 100 and 150 mg daily, respectively, for 5 consecutive days of a 28-day cycle. Idasanutlin was well tolerated; no dose-limiting toxicity was observed, but low-grade gastrointestinal toxicity was common. Overall response rate after 6 cycles was 58% (7 of 12) with idasanutlin monotherapy and 50% (2 of 4) with combination therapy. Median duration of response was 16.8 months (range, 3.5-26.7). Hematologic, symptomatic, pathologic, and molecular responses were observed. These data indicate that idasanutlin is a promising novel agent for PV; it is currently being evaluated in a global phase 2 trial. This trial was registered at www.clinicaltrials.gov as #NCT02407080.
Subject(s)
Antineoplastic Agents/administration & dosage , Polycythemia Vera/drug therapy , Pyrrolidines/administration & dosage , para-Aminobenzoates/administration & dosage , Administration, Oral , Adult , Aged , Aged, 80 and over , Antineoplastic Agents/adverse effects , Biomarkers , Female , Humans , Male , Middle Aged , Molecular Targeted Therapy , Mutation , Polycythemia Vera/diagnosis , Polycythemia Vera/etiology , Proto-Oncogene Proteins c-mdm2/antagonists & inhibitors , Proto-Oncogene Proteins c-mdm2/genetics , Proto-Oncogene Proteins c-mdm2/metabolism , Pyrrolidines/adverse effects , Treatment Outcome , para-Aminobenzoates/adverse effectsABSTRACT
PURPOSE: All trans-retinoic acid (ATRA) is successful in treating acute promyelocytic leukemia (APL) by inducing terminal differentiation-mediated cell death, but it has limited activity in non-APL acute myeloid leukemia (AML). We aim to improve ATRA therapy of AML by enhancing apoptosis through repression of the antiapoptotic proteins Bcl-2 and Mcl-1. EXPERIMENTAL DESIGN: APL and AML cell lines, as well as primary AML samples, were used to explore the mechanisms regulating differentiation and apoptosis during ATRA treatment. Stable transfection and gene silencing with siRNA were used to identify the key factors that inhibit apoptosis during induction of differentiation and drugs that accelerate apoptosis. RESULTS: In differentiation-responsive AML cells, ATRA treatment induces long-lasting repression of Bcl-2 while first upmodulating and then reducing the Mcl-1 level. The Mcl-1 level appears to serve as a gatekeeper between differentiation and apoptosis. During differentiation induction, activation of MEK/ERK and PI3K/Akt pathways by ATRA leads to activation of p90RSK and inactivation of glycogen synthase kinase 3ß (GSK3ß), which increase Mcl-1 levels by increasing its translation and stability. Sorafenib blocks ATRA-induced Mcl-1 increase by reversing p90RSK activation and GSK3ß inactivation, maintains the repressed Bcl-2 level, and enhances ATRA induced apoptosis in non-APL AML cell lines and in primary AML cells. CONCLUSIONS: Inhibition of Mcl-1 is required for apoptosis induction in ATRA differentiation-responsive AML cells. ATRA and sorafenib can be developed as a novel drug combination therapy for AML patients because this drug combination augments apoptosis by inhibiting Bcl-2 and Mcl-1.
Subject(s)
Leukemia, Myeloid, Acute/drug therapy , Leukemia, Promyelocytic, Acute/drug therapy , Myeloid Cell Leukemia Sequence 1 Protein/biosynthesis , Proto-Oncogene Proteins c-bcl-2/biosynthesis , Tretinoin/administration & dosage , Antineoplastic Combined Chemotherapy Protocols/administration & dosage , Apoptosis/drug effects , Cell Differentiation/drug effects , Gene Expression Regulation, Neoplastic/drug effects , Glycogen Synthase Kinase 3 beta/biosynthesis , HL-60 Cells , Humans , Leukemia, Myeloid, Acute/genetics , Leukemia, Myeloid, Acute/pathology , Leukemia, Promyelocytic, Acute/genetics , Leukemia, Promyelocytic, Acute/pathology , Myeloid Cell Leukemia Sequence 1 Protein/genetics , Niacinamide/administration & dosage , Niacinamide/analogs & derivatives , Phenylurea Compounds/administration & dosage , Phosphatidylinositol 3-Kinases/genetics , Proto-Oncogene Proteins c-bcl-2/genetics , RNA, Small Interfering/genetics , Sorafenib , Xenograft Model Antitumor AssaysABSTRACT
Myelofibrosis (MF) is characterized by cytopenias, constitutional symptoms, splenomegaly, and marrow histopathological abnormalities (fibrosis, increased microvessel density, and osteosclerosis). The microenvironmental abnormalities are likely a consequence of the elaboration of a variety of inflammatory cytokines generated by malignant megakaryocytes and monocytes. We observed that levels of a specific inflammatory cytokine, lipocalin-2 (LCN2), were elevated in the plasmas of patients with myeloproliferative neoplasms (MF > polycythemia vera or essential thrombocythemia) and that LCN2 was elaborated by MF myeloid cells. LCN2 generates increased reactive oxygen species, leading to increased DNA strand breaks and apoptosis of normal, but not MF, CD34(+) cells. Furthermore, incubation of marrow adherent cells or mesenchymal stem cells with LCN2 increased the generation of osteoblasts and fibroblasts, but not adipocytes. LCN2 priming of mesenchymal stem cells resulted in the upregulation of RUNX2 gene as well as other genes that are capable of further affecting osteoblastogenesis, angiogenesis, and the deposition of matrix proteins. These data indicate that LCN2 is an additional MF inflammatory cytokine that likely contributes to the creation of a cascade of events that results in not only a predominance of the MF clone but also a dysfunctional microenvironment.