RESUMO
Curly top disease, caused by beet curly top virus (BCTV), is among the most serious viral diseases affecting sugar beets in western USA. The virus is exclusively transmitted by the beet leafhopper (BLH, Circulifer tenellus) in a circulative and non-propagative manner. Despite the growing knowledge on virus-vector interactions, our understanding of the molecular interactions between BCTV and BLH is hampered by limited information regarding the virus impact on the vector and the lack of genomic and transcriptomic resources for BLH. This study unveils the significant impact of BCTV on both the performance and transcriptome response of BLHs. Viruliferous BLHs had higher fecundity than non-viruliferous counterparts, which was evident by upregulation of differentially expressed transcripts (DETs) associated with development, viability and fertility of germline and embryos in viruliferous insects. Conversely, most DETs associated with muscle movement and locomotor activities were downregulated in viruliferous insects, implying potential behavioural modifications by BCTV. Additionally, a great proportion of DETs related to innate immunity and detoxification were upregulated in viruliferous insects. Viral infection also induced notable alterations in primary metabolisms, including energy metabolism, namely glucosidases, lipid digestion and transport, and protein degradation, along with other cellular functions, particularly in chromatin remodelling and DNA repair. This study represents the first comprehensive transcriptome analysis for BLH. The presented findings provide new insights into the multifaceted effects of viral infection on various biological processes in BLH, offering a foundation for future investigations into the complex virus-vector relationship and potential management strategies for curly top disease.
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Beta vulgaris , Perfilação da Expressão Gênica , Hemípteros , Insetos Vetores , Doenças das Plantas , Animais , Hemípteros/virologia , Hemípteros/genética , Doenças das Plantas/virologia , Doenças das Plantas/genética , Insetos Vetores/virologia , Insetos Vetores/genética , Beta vulgaris/virologia , Transcriptoma , Geminiviridae/genética , Geminiviridae/fisiologia , Fertilidade/genéticaRESUMO
BACKGROUND: Cytomegalovirus (CMV) infection can significantly complicate and worsen the condition of acute severe ulcerative colitis (UC) patients. We aimed to explore the predictive risk factors to prevent and identify CMV infection at an early stage in acute UC patients. METHODS: A total of 115 moderate-to-severe active UC patients from 17 hospitals throughout China were enrolled. Active CMV infection was diagnosed by one of the following: CMV pp65 antigens, CMV IgM antibodies or CMV DNA. We identified the independent risk factors by multivariate analyses. RESULTS: A total of 64 of 115 active UC patients had active CMV infection. Compared to the non-CMV-infected patients, the CMV-infected patients had a tendency to be male and to exhibit abdominal pain; fever; oral ulcers; eosinopenia; low albumin, immunoglobulin (Ig) A, IgM, and IgG levels; increased high-sensitivity C-reactive protein (hsCRP) levels; hyponatremia; pancolonic lesions; initial onset type; severe activity; and glucocorticoid (high-dose) and immunosuppressive agent use (P < 0.05). In further multivariate analyses, the use of high-dose glucocorticoids (OR 13.55, 95% CI 2.49-73.61, P < 0.01) and immunosuppressive agents (OR 11.23, 95% CI 1.05-119.99, P = 0.04) were independent risk factors for CMV infection. A decrease eosinophil and albumin levels were risk factors for CMV infection. With every 0.1*10^9/L decrease in the peripheral blood eosinophil level or 1 g/L decrease in the serum albumin level, the risk for CMV infection in UC patients increased by 5.21-fold (1/0.192) or 1.19-fold (1/0.839), respectively. CONCLUSIONS: High-dose glucocorticoid and immunosuppressive agent treatment significantly increase the risk of CMV infection, and correcting eosinopenia and low albumin levels may help prevent CMV infection in UC patients.
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Colite Ulcerativa , Infecções por Citomegalovirus , Albuminas , China/epidemiologia , Colite Ulcerativa/complicações , Colite Ulcerativa/tratamento farmacológico , Citomegalovirus , Infecções por Citomegalovirus/complicações , DNA Viral , Humanos , Imunoglobulina A , MasculinoRESUMO
OBJECTIVE: To observe the effects and safety of quadruple regimens including domestically manufactured rabeprazole used as first line/initial therapy for Helicobacter pylori(H.pylori) eradication in gastritis and duodenal ulcer patients, and to investigate the effects of extended use of bismuth after the quadruple therapy on eradication of H. pylori. METHODS: From January to August 2013, 430 patients with chronic gastritis or duodenal ulcer who were confirmed as H. pylori positive in gastroscopy for upper gastrointestinal symptoms were enrolled from 12 centers in China for initial treatment using quadruple regimens for H. pylori eradication. The study was a prospective, multicenter, randomized double-blinded double-dummy parallel-controlled clinical trial. The 310 chronic gastritis patients were divided into 2 groups: group A1 was given quadruple regime (rabeprazole+ amoxicillin+ clarithromycin+ bismuth potassium citrate) for 10 days followed by bismuth-placebo for 21 days; group A2 was given the quadruple regimen for 10 days and then bismuth potassium citrate for 21 days. The duodenal ulcer patients were given the quadruple for 10 days, then rabeprazole for 14 days. All the patients took (13)C urea breath test to detect H. pylori 28 days after medicine withdrawal. RESULTS: Altogether 428 cases were enrolled and 404 completed the trial. The total eradication rate in the chronic gastritis patients was 85.1% (262/308, intention-to-treat (ITT)analysis), which was 81.7% (125/153, ITT) in the A1 group and 88.4% (137/155, ITT) in the A2 group; the eradication rate in the duodenal ulcer patients was 85.8% (103/120, ITT). No severe adverse effects were reported. The symptoms (pain, burning sensation, reflux, belching, nausea, and vomiting) improvement status was similar among A1 and A2 groups. CONCLUSIONS: The quadruple regimen using rabeprazole manufactured in China and administered for 10 days as first line/initial therapy in chronic gastritis and duodenal ulcer patients could achieve good H. pylori eradication rate. The extended use of bismuth after 10-day quadruple regimen might further improve the eradication rate. The regimens containing proton-pump inhibitor and bismuth may be well tolerated and safe in clinical application.
Assuntos
Úlcera Duodenal , Helicobacter pylori , Amoxicilina , Antiácidos , Bismuto , Testes Respiratórios , Quimioterapia Combinada , Gastrite , Refluxo Gastroesofágico , Gastroscopia , Infecções por Helicobacter , Humanos , Compostos Organometálicos , Estudos Prospectivos , Inibidores da Bomba de Prótons , UreiaRESUMO
BACKGROUND/AIMS: We aimed to observe the changes in the anti-Helicobacter pylori (Hp) serum antibodies to Hp virulence factors after eradication therapy and evaluate the potential application value of protein microarray in detecting Hp antibodies after eradication therapy. METHODOLOGY: A total of 107 Hp-positive patients with peptic ulcers (55) and chronic gastritis (52) were recruited. Serum antibodies to Hp urease (Ure), cytotoxin-associated protein (CagA), vacuolating cytotoxin (VacA), heat shock protein 60 (Hsp60), and anti-RdxA nitroreductase were measured. Four weeks after treatment, a 13C-urea breath test (13C- UBT) was applied to assess the Hp eradication state and to analyze correlations between the Hp eradication rate and the five antibodies. Six months after the therapy, protein microarray analysis was used to study the changes in these five serum antibodies. RESULTS: The overall Hp eradication rate was 86.0%There was no significant difference in the rate among the groups that tested positive and negative for the remaining four virulence factors. CONCLUSION: The disease type and serum anti-CagA antibody levels affect the therapeutic outcome of Hp eradication therapy. Protein microarray detection of Hp-related antibodies did not have significant application value for the long-term follow-up of Hp infection after eradication therapy.
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Antibacterianos/uso terapêutico , Anticorpos Antibacterianos/sangue , Infecções por Helicobacter/tratamento farmacológico , Helicobacter pylori/efeitos dos fármacos , Antagonistas dos Receptores H2 da Histamina/uso terapêutico , Análise Serial de Proteínas , Inibidores da Bomba de Prótons/uso terapêutico , Fatores de Virulência/imunologia , Adulto , Idoso , Antígenos de Bactérias/imunologia , Proteínas de Bactérias/imunologia , Testes Respiratórios , Doença Crônica , Úlcera Duodenal/tratamento farmacológico , Úlcera Duodenal/microbiologia , Feminino , Seguimentos , Gastrite/tratamento farmacológico , Gastrite/microbiologia , Infecções por Helicobacter/diagnóstico , Infecções por Helicobacter/microbiologia , Helicobacter pylori/imunologia , Helicobacter pylori/patogenicidade , Humanos , Masculino , Pessoa de Meia-Idade , Indução de Remissão , Úlcera Gástrica/tratamento farmacológico , Úlcera Gástrica/microbiologia , Fatores de Tempo , Resultado do Tratamento , Urease/imunologia , VirulênciaRESUMO
BACKGROUND/AIMS: The relationship of miR-181c, a potential tumor regulatory factor, with gastric cancer is not well studied. We have investigated this relationship in our study. METHODOLOGY: Paraffin-embedded tissue specimens from 103 gastric cancer patients were subjected to total RNA extraction. Reverse transcription real-time fluorescence quantitative PCR was used to detect miR-181c expression. Its relative expression was correlated with the patients' clinicopathological features. Also, the 5-year survival rate and median survival time were correlated with the miR-181c expression level. RESULTS: miR-181c expression was significantly and directly correlated with the degree of tumor differentiation, invasive depth and clinical stage. Moreover, lymph node metastasis was significantly related with higher miR-181c expression. However, miR-181c expression was not significantly correlated with gender, age, tumor location and distant metastasis. The 5-year overall survival rate and median survival time were significantly greater in patients with low expression of miR-181c. CONCLUSIONS: miR-181c expression level was significantly related to several clinicopathological features of gastric cancer; therefore, miR-181c probably plays a role in its development and progression. This relationship needs to be studied in detail, so that the potential use of miR-181c as a marker for gastric cancer and therapeutic target can be explored.
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MicroRNAs/fisiologia , Neoplasias Gástricas/patologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Progressão da Doença , Feminino , Humanos , Metástase Linfática , Masculino , MicroRNAs/análise , Pessoa de Meia-Idade , Prognóstico , Neoplasias Gástricas/genética , Neoplasias Gástricas/mortalidadeRESUMO
Human cellular reprogramming to induced pluripotency is still an inefficient process, which has hindered studying the role of critical intermediate stages. Here we take advantage of high efficiency reprogramming in microfluidics and temporal multi-omics to identify and resolve distinct sub-populations and their interactions. We perform secretome analysis and single-cell transcriptomics to show functional extrinsic pathways of protein communication between reprogramming sub-populations and the re-shaping of a permissive extracellular environment. We pinpoint the HGF/MET/STAT3 axis as a potent enhancer of reprogramming, which acts via HGF accumulation within the confined system of microfluidics, and in conventional dishes needs to be supplied exogenously to enhance efficiency. Our data suggest that human cellular reprogramming is a transcription factor-driven process that it is deeply dependent on extracellular context and cell population determinants.
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Células-Tronco Pluripotentes Induzidas , Humanos , Células-Tronco Pluripotentes Induzidas/metabolismo , Reprogramação Celular , Regulação da Expressão Gênica , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo , Células CultivadasRESUMO
OBJECTIVE: To explore the effects of 7-day quadruple regimen as the first-line therapy strategy for Helicobacter pylori(H. pylori)infection and compare the eradication rate of ilaprazole versus esoprazole-based regimen. METHODS: A total of 440 patients with H. pylori infection, who had never received H. pylori eradication treatment, were enrolled from 10 domestic hospitals from October 2010 to July 2011. Diagnosed as chronic gastritis or duodenal ulcer according to their endoscopic examination results, they were randomized into ilaprazole and(or) esoprazole-based bismuth-containing quadruple regimen group with amoxicillin and clarithromycin (n = 110 each). After a 7-day eradication treatment, all patients with duodenal ulcer received PPI (ilaprazole and(or) esoprazole) treatment for 14 days and (13)C urea breath test was performed at least 28 days after the end of therapy. The patients with failed eradication treatment underwent endoscopy examination and biopsy. H. pylori culture and detection of antibiotic-resistant genes were also performed. RESULTS: In gastritis patients, the eradication rate (per-protocol, PP value) were 78.2% (79/101) and 82.0% (82/100) in ilaprazole and esoprazole groups (P = 0.50) while the (intention-to-treat) ITT value of eradication rate were 71.8% (79/110) and 74.5% (82/110) in ilaprazole and esoprazole groups respectively (P = 0.65). And there was no statistical difference (P > 0.05). In duodenal patients, the eradication rate (PP) were 92.1% (93/101) and 91.4% (96/105) in ilaprazole and esoprazole group (P = 0.86) while the ITT value of eradication rate were 84.5% (93/110) and 87.3% (96/110) in ilaprazole and esoprazole groups respectively (P = 0.56). And no significant difference existed between two groups in gastritis and duodenal ulcer patients (P > 0.05). In total, the eradication rate was 80.1% (161/201) (PP) and 73.2% (161/220) (ITT), 91.7% (189/206) (PP) and 85.9% (189/220) (ITT) in chronic gastritis and duodenal ulcer patients respectively. The symptomatic improvements of stomachache, burning, belching and nausea remained almost unchanged. No severe side effect was observed. The point mutations for clarithromycin resistance were detected in all 53 H. pylori strains (100%) isolated from the patients with failed eradication treatment. CONCLUSIONS: The eradication rate of PPI based bismuth-containing quadruple regimen as the first-line treatment is satisfactory in chronic gastritis and duodenal ulcer patients. No significant difference exists between the effects of ilaprazole and esoprazole-based groups. And the treatment failure may be attributed mainly to the clarithromycin resistance of H. pylori.
Assuntos
2-Piridinilmetilsulfinilbenzimidazóis/uso terapêutico , Antiulcerosos/uso terapêutico , Infecções por Helicobacter/tratamento farmacológico , Omeprazol/uso terapêutico , 2-Piridinilmetilsulfinilbenzimidazóis/administração & dosagem , Adolescente , Adulto , Idoso , Antiulcerosos/administração & dosagem , China , Quimioterapia Combinada , Feminino , Helicobacter pylori , Humanos , Masculino , Pessoa de Meia-Idade , Omeprazol/administração & dosagem , Adulto JovemRESUMO
OBJECTIVE: To explore the efficacy of Jinghuaweikang capsules plus triple therapy (LACJ) in treatment of Helicobacter pylori (H. pylori) associated gastritis or duodenal ulcer, compare it with bismuth-containing quadruple therapy (LACB) and standard triple therapy (LAC) and analyze the antibiotic sensitivity of gastric mucosal H. pylori strains from the failed patients. METHODS: A total of 565 patients with H. pylori infection were recruited from 11 hospitals from January 2010 to June 2011. There were 336 males and 229 females. They underwent gastroendoscopy examination due to upper gastrointestinal symptoms and had never received H. pylori eradication therapies. Duodenal ulcer patients were divided randomly into LACJ therapy group, LACB therapy group and LAC therapy group while gastritis patients LACJ therapy group and LACB therapy group. Group LAC received lansoprazole 30 mg + amoxicillin 1000 mg + clarithromycin 500 mg, twice a day, for 7 d (d1-7). Group LACJ: LAC therapy plus Jinghuaweikang, 3 capsules, twice a day, for 7 d (d1-7) then Jinghuaweikang, 3 capsules, twice a day, for 14 d (d8-21). Group LACB: LAC plus bismuth potassium citrate 220 mg, twice a day, for 7 d (d1-7) and then bismuth potassium citrate 220 mg, twice a day, for 14 d (d8-21). All duodenal ulcer patients received lansoprazole (30 mg, once a day) for 14 days after the first 7-day of treatment (d 8-21). At least 28 days after the end of treatment, all patients underwent (13)C urea breath test. Gastric mucosa was collected under endoscopy from the failed patients. The detection technique of gene chip was employed to detect antibiotics resistant gene from mucosa. RESULTS: The eradication rates of duodenal ulcer patients in groups LACJ, LACB and LAC were as follows: per-protocol (PP), 80.2% (77/96), 89.9% (89/99) and 72.2% (70/97) (P = 0.007), intention-to-treat (ITT), 78.6% (77/98), 88.1% (89/101) and 70.0% (70/100) (P = 0.007). No statistical differences existed between groups LACJ and LACB or LAC (all P > 0.05). But there were statistical differences between groups LACB and LAC (both P = 0.002). The eradication rates of PP and ITT of chronic gastritis patients in groups LACJ and LACB were as follows: 75.8% (97/128), 74.6% (97/130) vs 83.8% (109/130), 80.1% (109/136) (both P > 0.05). The symptomatic improvements of abdominal pain, burning and acid reflux of duodenal ulcer patients in group LACJ were higher than those in groups LACB and LAC. There were statistical differences between groups LACJ and LAC (all P < 0.05). The symptomatic improvements of bloating and belching for chronic gastritis patients in group LACJ were higher than those of group LACB. But no significant difference existed between two groups (all P > 0.05). Sixty samples of gastric mucosa were collected from the failed patients. The detection rates of antibiotic-resistant gene to clarithromycin and amoxicillin were 60.0% (36/36) and 18.3% (11/60) respectively. CONCLUSIONS: The efficacy of LACJ for the treatment of H. pylori infection patients is similar to LACB and superior to LAC. And the symptomatic improvement of patients is better than the other two regimens. The main cause of treatment failure is antibiotic resistance of H. pylori strains.
Assuntos
Medicamentos de Ervas Chinesas/uso terapêutico , Úlcera Duodenal/tratamento farmacológico , Gastrite/tratamento farmacológico , Infecções por Helicobacter/tratamento farmacológico , Adulto , Farmacorresistência Bacteriana , Úlcera Duodenal/microbiologia , Feminino , Gastrite/microbiologia , Infecções por Helicobacter/microbiologia , Helicobacter pylori , Humanos , Masculino , Pessoa de Meia-Idade , Estudos ProspectivosRESUMO
BACKGROUND: The outbreak of the coronavirus disease 2019 (COVID-19) caused by the severe acute respiratory syndrome coronavirus 2 has been the most important clinical challenge worldwide since January 2020. COVID-19 inactivated vaccines play a crucial role in reducing the rates of morbidity and mortality. CASE SUMMARY: We presented a 48-year-old woman from Haidian District, Beijing, China who developed ischemic colitis after receiving the second dose of COVID-19 inactivated vaccine. Computed tomography of the abdomen showed edema and bowel wall thickening with hypodensity in the sigmoid colon and descending colon. Colonoscopy revealed hyperemia, edema and erosion of the mucosa with superficial ulceration and a yellow-white coating at the descending colon and sigmoid colon. The symptoms were relieved after 1 wk of receiving pinaverium bromide (50 mg, tid) and aspirin enteric-coated tablets (0.1 g, qd). CONCLUSION: The possible occurrence of ischemic colitis should be considered after administration of the COVID-19 inactivated vaccines.
RESUMO
BACKGROUND: Phosphorylated proteins are known to be present in multiple body fluids in normal conditions, and abnormally accumulated under some pathological conditions. The biological significance of their role in the extracellular space has started being elucidated only recently, for example in bone mineralization, neural development, and coagulation. Here, we address some criticalities of conventional culture systems for the study of the extracellular regulation of phosphorylation. METHODS: We make use of microfluidics to scale-down the culture volume to a size comparable to the interstitial spaces occurring in vivo. The phosphoprotein content of conditioned media was analyzed by a colorimetric assay that detects global phosphorylation. RESULTS: We found that miniaturization of the culture system increases phosphoprotein accumulation. Moreover, we demonstrated that in conventional culture systems dilution affects the extent of the phosphorylation reactions occurring within the extracellular space. On the other hand, in microfluidics the phosphorylation status was not affected by addition of adenosine triphosphate (ATP) and FAM20C Golgi Associated Secretory Pathway Kinase (FAM20C) ectokinase, as if their concentration was already not limiting for the phosphorylation reaction to occur. CONCLUSIONS: The volume of the extracellular environment plays a role in the process of extracellular phosphorylation due to its effect on the concentration of substrates, enzymes and co-factors. GENERAL SIGNIFICANCE: Thus, the biological role of extracellular phosphoregulation may be better appreciated within a microfluidic culture system.
Assuntos
Calcificação Fisiológica , Fosfoproteínas , Trifosfato de Adenosina/metabolismo , Complexo de Golgi/metabolismo , Fosfoproteínas/metabolismo , FosforilaçãoRESUMO
Nuclear deformation is an essential phenomenon allowing cell migration and can be observed in association with pathological conditions such as laminopathies, neurodegenerative disorders and diabetes. Abnormal nuclear morphologies are a hallmark of cancer progression and nuclear deformability is a necessary feature for metastatic progression. Nevertheless, the cellular processes and the key molecular components controlling nuclear shape are poorly understood, in part due to a limited availability of assays that allow high-throughput screening of nuclear morphology-phenotypes. In this study, we explore the application of micropillared substrates as the basis for a phenotypic screening platform aimed at identifying novel determinants of nuclear morphology. We designed PDMS substrates to maximize simplicity in image acquisition and analyses, and in a small-scale screening of inhibitors targeting chromatin-modifying enzymes, we identify histone deacetylation as cellular process involved in nuclear deformation. With increasingly specific targeting approaches, we identify HDAC2 as a novel player in controlling nuclear morphology through gene transcription repression. This study shows the effectiveness of micropillar-based substrates to act as phenotypic drug screening platforms and opens a new avenue in the identification of genes involved in determining the nuclear shape.
Assuntos
Histona Desacetilase 2 , Neoplasias , Cromatina/genética , Cromatina/metabolismo , Avaliação Pré-Clínica de Medicamentos , Histona Desacetilase 2/genética , Histona Desacetilase 2/metabolismo , Humanos , FenótipoRESUMO
The establishment of in vitro naive human pluripotent stem cell cultures opened new perspectives for the study of early events in human development. The role of several transcription factors and signaling pathways have been characterized during maintenance of human naive pluripotency. However, little is known about the role exerted by the extracellular matrix (ECM) and its three-dimensional (3D) organization. Here, using an unbiased and integrated approach combining microfluidic cultures with transcriptional, proteomic, and secretome analyses, we found that naive, but not primed, hiPSC colonies are characterized by a self-organized ECM-rich microenvironment. Based on this, we developed a 3D culture system that supports robust long-term feeder-free self-renewal of naive hiPSCs and also allows direct and timely developmental morphogenesis simply by modulating the signaling environment. Our study opens new perspectives for future applications of naive hiPSCs to study critical stages of human development in 3D starting from a single cell.
Assuntos
Células-Tronco Pluripotentes Induzidas , Células-Tronco Pluripotentes , Humanos , Proteômica , Matriz Extracelular , MorfogêneseRESUMO
OBJECTIVE: To contrast the effect of rebamipide with proton pump inhibitors (PPIs) versus PPIs alone for the treatment of endoscopic submucosal dissection (ESD-) induced ulcers. METHODS: PubMed, Embase, the Cochrane library, the WanFang database, and China National Knowledge Infrastructure (CNKI) were searched to identify studies that met the inclusion criteria. RESULTS: Nine randomized controlled trials (RCTs) were recognized, including 1170 patients. In general, rebamipide plus PPIs acted better than PPIs alone against ESD-induced ulcers at four weeks (RR = 1.42, 95% CI: 1.13-1.78, P = 0.003) but showed no significant differences at eight weeks (RR = 1.03, 95% CI: 0.97-1.10, P = 0.315). The use of rebamipide plus PPIs was superior to PPIs alone for ESD-induced ulcers greater than 20 mm in size (20-40 mm: RR = 1.98, 95% CI: 1.22-3.23, P = 0.006; >40 mm: RR = 5.14, 95% CI: 1.49-17.74, P = 0.010). In addition, rebamipide plus PPI therapy was discovered to be significantly more effective than PPIs alone for lower ESD-induced ulcers (RR = 1.82, 95% CI: 1.04-3.20, P = 0.037). There were no significant differences between the treatment groups with the ulcer reduction rate. CONCLUSION: Evidences now available show rebamipide plus PPIs is practical for protecting against ESD-induced ulcers at four weeks but not at eight weeks, especially large ulcers (>20 mm). However, we still need more high-quality RCTs in the future to supplement our conclusions.
Assuntos
Alanina/análogos & derivados , Antiulcerosos/uso terapêutico , Ressecção Endoscópica de Mucosa/efeitos adversos , Inibidores da Bomba de Prótons/uso terapêutico , Quinolonas/uso terapêutico , Úlcera Gástrica/tratamento farmacológico , Alanina/uso terapêutico , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Úlcera Gástrica/etiologiaRESUMO
Nuclear shape modulates cell behavior and function, while aberrant nuclear morphologies correlate with pathological phenotype severity. Nevertheless, functions of specific nuclear morphological features and underlying molecular mechanisms remain poorly understood. Here, we investigate a nucleus-intrinsic mechanism driving nuclear lobulation and segmentation concurrent with granulocyte specification, independently from extracellular forces and cytosolic cytoskeleton contributions. Transcriptomic regulation of cholesterol biosynthesis is equally concurrent with nuclear remodeling. Its putative role as a regulatory element is supported by morphological aberrations observed upon pharmacological impairment of several enzymatic steps of the pathway, most prominently the sterol ∆14-reductase activity of laminB-receptor and protein prenylation. Thus, we support the hypothesis of a nuclear-intrinsic mechanism for nuclear shape control with the putative involvement of the recently discovered GGTase III complex. Such process could be independent from or complementary to the better studied cytoskeleton-based nuclear remodeling essential for cell migration in both physiological and pathological contexts such as immune system function and cancer metastasis.
Assuntos
Citoesqueleto/metabolismo , Granulócitos/metabolismo , Prenilação de Proteína/genética , Células HL-60 , Humanos , Modelos MolecularesRESUMO
Injury to the skin from severe burns can cause debilitating physical and psychosocial distress to the patients. Upon healing, deep dermal burns often result in devastating hypertrophic scar formation. For many decades, stem cell-based therapies have shown significant potential in improving wound healing. However, current cell delivery methods are often insufficient to maintain cell viability in a harmful burn wound environment to promote skin regeneration. In this study, we developed an enhanced approach to deliver adipose-derived stem cells (ASCs) for the treatment of burn wounds, using an in-situ-formed hydrogel system comprised of a hyperbranched poly(ethylene glycol) diacrylate (HB-PEGDA) polymer, a commercially available thiol-functionalized hyaluronic acid (HA-SH) and a short RGD peptide. Stable hydrogels with tunable swelling and mechanical properties form within five minutes under physiological conditions via the Michael-type addition reaction. Combining with RGD peptide, as a cell adhesion motif, significantly alters the cellular morphology, enhances cell proliferation, and increases the paracrine activity of angiogenesis and tissue remodeling growth factors and cytokines. Bioluminescence imaging of luciferase+ ASCs indicated that the hydrogel protected the implanted cells from the harmful wound environment in burns. Hydrogel-ASC treatment significantly enhanced neovascularization, accelerated wound closure and reduced the scar formation. Our findings suggest that PEG-HA-RGD-based hydrogel provides an effective niche capable of augmenting the regenerative potential of ASCs and promoting burn wound healing. STATEMENT OF SIGNIFICANCE: Burn injury is one of the most devastating injures, and patients suffer from many complications and post-burn scar formation despite modern therapies. Here, we designed a conformable hydrogel-based stem cell delivery platform that allows rapid in-situ gelation upon contact with wounds. Adipose-derived stem cells were encapsulated into a PEG-HA-RGD hydrogels. Introducing of RGD motif significantly improved the cellular morphology, proliferation, and secretion of angiogenesis and remodeling cytokines. A deep second-degree burn murine model was utilized to evaluate in-vivo cell retention and therapeutic effect of the hydrogel-ASC-based therapy on burn wound healing. Our hydrogel remarkably improved ASCs viability in burn wounds and the hydrogel-ASC treatment enhanced the neovascularization, promoted wound closure, and reduced scar formation.
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Ácido Hialurônico , Hidrogéis , Adipócitos , Tecido Adiposo , Animais , Humanos , Camundongos , Células-TroncoRESUMO
Osmotic stress, caused by drought, salinity, or PEG (polyethylene glycol), is one of the most important abiotic factors that hinder plant growth and development. In Arabidopsis, more than 100 R2R3-MYB transcription factors (TFs) have been identified, and many of them are involved in the transcriptional regulation of a variety of biological processes related to growth and development, as well as responses to biotic and abiotic stresses. However, the MYB TF involving in both plant development and stress response has rarely been reported. We report here that Arabidopsis AtMYB109, a R2R3-MYB TF, functions as a negative regulator of stomatal closure under osmotic stress as well as of pollen tube elongation. Under PEG-induced osmotic stress, whole leaves of AtMYB109-OXs were intensely wilted, while leaves of the wild-type (WT) and myb109 were weakly affected. Moreover, we confirmed that the wilting in AtMYB109-OXs was more severe than in WT and myb109 under drought conditions, and that after re-watering, WT and myb109 plants promptly recovered, while AtMYB109-OXs failed to survive. In addition, stomatal closure was delayed in the AtMYB109-OXs compared to the WT and myb109. However, proline content and the expression of stress-induced and proline synthesis genes were higher in the overexpression lines than in WT and myb109. Then, we observed that the expression of ICS1, a key gene in SA biosynthesis, was greatly suppressed in AtMYB109-OXs. In addition, we found that AtMYB109 expression gradually increased until the flowers were fully opened and thereafter dramatically decreased during silique development. The pollen tube growth was significantly suppressed in AtMYB109-OXs compared to the WT and myb109. Using EMSA and ChIP-qPCR, we confirmed that AtMYB109 bound to the promoter of RABA4D, a gene encoding a pollen development regulator. Taken together, we suggest the delayed stomatal closing and vulnerable phenotypes in the AtMYB109-OXs under osmotic stress are possibly directly or indirectly associated with a SA-mediated mechanism, and that AtMYB109 suppresses RABA4D that modulates pollen tube growth.
Assuntos
Arabidopsis/crescimento & desenvolvimento , Arabidopsis/genética , Regulação da Expressão Gênica de Plantas/fisiologia , Estômatos de Plantas/genética , Estômatos de Plantas/fisiologia , Estresse Fisiológico/fisiologia , Flores/crescimento & desenvolvimento , Flores/metabolismo , Genes de Plantas , Pressão Osmótica/fisiologia , Folhas de Planta/crescimento & desenvolvimento , Folhas de Planta/metabolismo , Raízes de Plantas/crescimento & desenvolvimento , Raízes de Plantas/metabolismoRESUMO
Skeletal muscle decellularization allows the generation of natural scaffolds that retain the extracellular matrix (ECM) mechanical integrity, biological activity, and three-dimensional (3D) architecture of the native tissue. Recent reports showed that in vivo implantation of decellularized muscles supports muscle regeneration in volumetric muscle loss models, including nervous system and neuromuscular junctional homing. Since the nervous system plays pivotal roles during skeletal muscle regeneration and in tissue homeostasis, support of reinnervation is a crucial aspect to be considered. However, the effect of decellularized muscles on reinnervation and on neuronal axon growth has been poorly investigated. Here, we characterized residual protein composition of decellularized muscles by mass spectrometry and we show that scaffolds preserve structural proteins of the ECM of both skeletal muscle and peripheral nervous system. To investigate whether decellularized scaffolds could per se attract neural axons, organotypic sections of spinal cord were cultured three dimensionally in vitro, in presence or in absence of decellularized muscles. We found that neural axons extended from the spinal cord are attracted by the decellularized muscles and penetrate inside the scaffolds upon 3D coculture. These results demonstrate that decellularized scaffolds possess intrinsic neurotrophic properties, supporting their potential use for the treatment of clinical cases where extensive functional regeneration of the muscle is required.
Assuntos
Matriz Extracelular/metabolismo , Imageamento Tridimensional/métodos , Músculo Esquelético/metabolismo , Proteômica/métodos , Engenharia Tecidual/métodos , Animais , Feminino , Humanos , Masculino , RatosRESUMO
OBJECTIVE: To investigate the expression of Abl-interacting protein 1 (ABI1) in normal gastric mucosal cell line GES-1 and gastric cancer cell line AGS, and the effects of ABI1 gene overexpression upon the proliferation of human gastric cancer cell AGS in vitro. METHODS: Firstly the ABI1 expression in GES-1 and AGS cells were identified by immunohistochemistry, immunofluorescence, RT-PCR, real-time PCR and Western blot. Secondly human gastric cancer cell line AGS was cultured and transfected with recombinant MSCV-GFP-ABI1 plasmid or blank plasmid MSCV-GPF. Real-time PCR and Western blot were used to detect the mRNA and protein expression of ABI1. And lastly the cell proliferation was detected by CCK-8 assay. RESULTS: ABI1 was expressed both in normal gastric mucosal cell line GES-1 and in gastric cancer cell line AGS. Compared to GES-1 cells, the ABI1 expression in AGS cells was lowered significantly. There were no significant differences in the ABI1 mRNA and protein expression between the AGS and AGS-MSCV-GFP groups. Compared to those of the AGS group, the ABI1 mRNA expression levels of the AGS-MSCV-GFP-ABI1 group increased by 1.87 times (P = 0.002). The protein expression levels of the AGS-MSCV-GFP-ABI1 group were remarkably higher than those of the AGS and AGS-MSCV-GFP groups (P = 0.002). CCK-8 assay showed that there were no significant differences in the proliferation rates at different time points between the AGS and AGS-MSCV-GFP groups. However, the proliferation rates at the time points of 24, 48, 72 and 96 hours of the AGS-MSCV-GFP-ABI1 were 1.46 +/- 0.31, 4.75 +/- 0.12, 6.62 +/- 0.32 and 8.96 +/- 0.27 respectively. And they were significantly lower than the proliferation rates of the AGS and AGS-MSCV-GFP groups (P < 0.01). CONCLUSION: ABI1 gene is down-regulated in gastric cancer cells. The ABI1 overexpression effectively inhibits the proliferation in human gastric cancer cell lines. It suggests that ABI1 may be involved in gastric cancer pathogenesis by regulating the proliferation of gastric carcinomas cells.
Assuntos
Proteínas Adaptadoras de Transdução de Sinal/metabolismo , Proliferação de Células , Proteínas do Citoesqueleto/metabolismo , Mucosa Gástrica/patologia , Neoplasias Gástricas/metabolismo , Neoplasias Gástricas/patologia , Apoptose , Linhagem Celular Tumoral , Mucosa Gástrica/citologia , Mucosa Gástrica/metabolismo , Humanos , TransfecçãoRESUMO
OBJECTIVE: To evaluate the diagnostic value of antineutrophil cytoplasmic antibodies (ANCA) in patients with ulcerative colitis (UC). METHODS: Articles related to diagnosis of UC by ANCA test, published before November 2007, were retrieved in the databanks such as Chinese BioMedical Disc, Chinese Medical Current Contents, China National Knowledge Infrastructure, and VIP databank. Related journals were searched manually. Meta-analysis was performed by summary receiver operating characteristic curve recommended by Diagnostic and Screening Group of the Cochrane Collaboration Web with the parameters such as sensitivity, specificity, accuracy, predictive values and likelihood ratio. RESULTS: Totally 10 studies including 381 patients met the inclusion criteria. Meta-analysis showed that the sensitivity and specificity of ANCA to the diagnosis of UC were 52.2% and 99.0% respectively. The positive and negative predictive values (PPV and NPV) were 98.0% and 68.7% respectively. Meanwhile, the positive and negative likelihood ratio (+LR and -LR) were 52.2 and 0.5, respectively. CONCLUSIONS: ANCA is an immunological index related to UC. Positive ANCA helps diagnose UC, but is not sensible enough to screen UC.
Assuntos
Anticorpos Anticitoplasma de Neutrófilos , Colite Ulcerativa/diagnóstico , Adolescente , Adulto , Idoso , Anticorpos Anticitoplasma de Neutrófilos/imunologia , Colite Ulcerativa/imunologia , Humanos , Pessoa de Meia-Idade , Neutrófilos/citologia , Periplasma/imunologia , Valor Preditivo dos Testes , Sensibilidade e EspecificidadeRESUMO
The pollen wall is a specialized extracellular cell wall matrix that surrounds male gametophytes and plays an essential role in plant reproduction. Uncovering the mechanisms that control the synthesis and polymerization of the precursors of pollen wall components has been a major research focus in plant biology. We review current knowledge on the genetic and biochemical mechanisms underlying pollen wall development in eudicot model Arabidopsis thaliana and monocot model rice (Oryza sativa), focusing on the genes involved in the biosynthesis, transport, and assembly of various precursors of pollen wall components. The conserved and divergent aspects of the genes involved as well as their regulation are addressed. Current challenges and future perspectives are also highlighted.