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1.
J Transl Med ; 17(1): 278, 2019 Aug 22.
Artigo em Inglês | MEDLINE | ID: mdl-31438987

RESUMO

BACKGROUND: Calcium oxalate (CaOx), the major constituent of most kidney stones, induces inflammatory infiltration and injures renal tubular cells. However, the role of γδT cells in CaOx-mediated kidney injury remains unclear. Therefore, this study investigated the distribution of intrarenal γδT cells and T cell receptor δ (TCRδ) immune repertoires in response to interactions with CaOx crystals. METHODS: CaOx crystal mouse model was established by glyoxylate injection. Flow cytometer was used to analyze the expression of CD69 and IL-17 from intrarenal γδT cells. Furthermore, TCR immune repertoire sequencing (IR-Seq) was used to monitor the profile of the TCRδ immune repertoire. RESULTS: Our results indicated that CaOx crystals lead to obvious increases in the expression and activation of intrarenal γδT cells. In TCRδ immune repertoire, the majority of V/J gene and V-J/V-D-J combination segments, barring individual exceptions, were similar between kidneys with CaOx formation and control kidneys. Impressively, high complementarity determining region 3 (CDR3) diversity was observed in response to CaOx crystal formation along with distinct CDR3 distribution and abundance. CONCLUSION: Our work suggests the presence of aberrant γδT cell activation and reconstitution of the TCRδ immune repertoire in response to CaOx crystal deposition.

2.
J Bacteriol ; 2019 Jul 01.
Artigo em Inglês | MEDLINE | ID: mdl-31262840

RESUMO

Pseudomonas stutzeri A1501 is a versatile nitrogen-fixing bacterium capable of living in diverse environments and coping with various oxidative stresses. NfiS, a regulatory noncoding RNA (ncRNA) involved in the control of nitrogen fixation in A1501, was previously shown to be required for optimal resistance to H2O2; however, the precise role of NfiS and the target genes involved in the oxidative stress response is entirely unknown. In this work, we systematically investigated the NfiS-based mechanisms underlying the response of this bacterium to H2O2 at the cellular and molecular levels. A mutant strain carrying a deletion of nfiS showed significant downregulation of oxidative response genes, especially katB, a catalase gene, and oxyR, an essential regulator for transcription of catalase genes. Secondary structure prediction revealed two binding sites in NfiS for katB mRNA. Complementation experiments using truncated nfiS genes showed that each of two sites is functional, but not sufficient, for NfiS-mediated regulation of oxidative stress resistance and nitrogenase activities. Microscale thermophoresis assays further indicated direct base-pairing between katB mRNA and NfiS at both sites 1 and 2, thus enhancing the half-life of the transcript. We also demonstrated that katB expression is dependent on OxyR and that both OxyR and KatB are essential for optimal oxidative stress resistance and nitrogenase activities. H2O2 at low concentrations was detoxified by KatB, leaving O2 as a by-product to support nitrogen fixation under O2-insufficient conditions. Moreover, our data suggest that the direct interaction between NfiS and katB mRNA is a conserved and widespread mechanism among P. stutzeri strains.IMPORTANCE Protection against oxygen damage is crucial for survival of nitrogen-fixing bacteria due to the extreme oxygen sensitivity of nitrogenase. This work exemplifies how the small ncRNA, NfiS, coordinates oxidative stress response and nitrogen fixation via base pairing with katB mRNA and nifK mRNA. Hence, NfiS acts as a molecular link to coordinate the expression of genes involved in oxidative stress response and nitrogen fixation. Our study provides the first insight into the biological functions of NfiS in oxidative stress regulation, and adds a new regulation level to the mechanisms that contribute to the oxygen protection of the MoFe nitrogenase.

3.
Eur J Pharmacol ; 857: 172457, 2019 Aug 15.
Artigo em Inglês | MEDLINE | ID: mdl-31202804

RESUMO

Short-chain fatty acids (SCFAs) are produced by the fermentation of dietary fiber by the gut microbiota and are beneficial to the health of the body. Insufficient SCFAs productions are associated with type 2 diabetes (T2D). We used a long-term high-fat diet to simulate the pathogenesis of T2D and studied the effects of baicalin on gut microbiota and metabolites in mice as well as its mechanism, providing a theoretical basis for the treatment of T2D. Baicalin groups were given 200 mg/kg/day, and control groups were given an equal volume of 0.5% sodium carboxymethyl cellulose solution for 15 weeks. 16S rRNA amplicon pyrosequences was performed to evaluate the gut microbiota composition, and gas chromatography was used to detect SCFAs in stool samples in the different experimental groups. The abundance of gut microbiota in the high-fat model group was altered, and was associated with a decreased production of SCFAs. The microbiota abundance of the baicalin group was closer to that of the control group, increasing the population of SCFA-producing bacteria spp and improving metabolic syndrome, including abnormal glucose and lipid metabolism caused by a high-fat diet. Baicalin may improve abnormalities in glycolipid metabolism by affecting the production of SCFAs.

4.
Appl Environ Microbiol ; 85(14)2019 Jul 15.
Artigo em Inglês | MEDLINE | ID: mdl-31076427

RESUMO

Expression of nitrogenase genes (nifHDK) is strictly regulated at both transcriptional and posttranscriptional levels. Efficient nitrogenase activity requires maintaining sufficient levels of nif mRNAs, yet the underlying mechanism is not fully understood due to its complexity. We have previously shown that a novel regulatory noncoding RNA (ncRNA), NfiS, optimizes nitrogen fixation through targeting nifK mRNA in Pseudomonas stutzeri A1501. Here, we report the identification and characterization of a second ncRNA inducible under nitrogen fixation conditions (nitrogen-free and microaerobic conditions), termed NfiR (for nitrogen fixation condition-inducible ncRNA), the expression of which is dependent on two global regulators, NtrC and Hfq. Comparative phenotypic and proteomic analyses of an nfiR mutant identify a role of NfiR in regulating the expression of nitrogenase genes. Further microscale thermophoresis and genetic complementation showed that an 11-nucleotide (nt) sequence in the stem-loop structure of NfiR (nucleotides 12 to 22) pairs with its counterpart in the coding region of nifD mRNA (nucleotides 1194 to 1207) by eight nucleotides. Significantly, deletion of nfiR caused a 60% reduction of nitrogenase activity, and the half-life of nifD mRNA was reduced from 20 min for the wild type to 15 min for the ΔnfiR mutant. With regard to nitrogenase activity and stability of the nifD and nifK transcripts, phenotypes were more severe for the double deletion mutant lacking nfiR and nfiS, suggesting that NfiR, in concert with NfiS, optimizes nitrogenase production at the posttranscriptional level.IMPORTANCE Biological nitrogen fixation is an energy-expensive process requiring the hydrolysis of 16 ATPs. Consequently, the expression of nif genes is highly regulated at both transcriptional and posttranscriptional levels through complex regulatory networks. Global regulation involves a number of regulatory proteins, such as the nif-specific activator NifA and the global nitrogen regulator NtrC, as well as various regulatory ncRNAs. We show that the two P. stutzeri ncRNAs, namely NfiS and NfiR (for nitrogen fixation condition-inducible ncRNA), optimize nitrogen fixation and environmental stress responses. NfiS and NfiR respond differently to various environmental signals and differ in their secondary structures. In addition, the two ncRNAs target the mRNAs of nifK and nifD, respectively. Such ncRNA-based posttranscriptional regulation of nitrogenase expression might be an evolved survival strategy, particularly in nitrogen-limiting environments. This study not only highlights the significant roles of regulatory ncRNAs in the coordination and fine tuning of various physiological processes but also provides a new paradigm for posttranscriptional regulation in nitrogen-fixing bacteria.

5.
Chin J Cancer Res ; 29(3): 196-203, 2017 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-28729770

RESUMO

OBJECTIVE: Detection rate and isolation yield of circulating tumor cell (CTC) are low in squamous cell carcinoma of head and neck (SCCHN) with in vitro approaches due to limited sample volumes. In this study, we applied the CellCollector to capture CTC in vivo from peripheral blood. METHODS: In total, the study included 22 cases with 37 times of detection. All of the patients were newly diagnosed with locally advanced or metastatic SCCHN, including laryngocarcinoma (40.9%, 9/22) and hypopharyngeal carcinoma (59.1%, 13/22). All patients received CTC analysis before treatment. Three patients received induction chemotherapy. Sixteen patients received surgical therapy, of which 13 patients received postoperative detection. Two patients received both induction chemotherapy and surgery treatment. Patients underwent two successive CellCollector applications 24 h before and 7 d after surgical therapy. Nine healthy volunteers were enrolled as the control group. Epidermal growth factor receptor variant type III (EGFRVIII) expression was analyzed with fluorescent dye labeled antibody. RESULTS: With CellCollector isolation, 72.7% (16/22) of the patients were positive for ≥1 CTC (CTC; range, 1-17 cells) before treatments and 46.7% (7/15) of patients were CTC positive for ≥1 CTC (CTC; range, 1-29 cells) after surgical therapy. Moreover, the detection rate of CellCollector (82.4%, 14/17; CTC count range, 0-17) in advanced SCCHN (stage III-IV) was much higher than that in early stages (stage I-II, 40.0%, 2/5; CTC count range, 0-2) (P<0.05). EGFRVIII expression of CTC was also analyzed with fluorescence staining. One CTCEGFRVIII-positive patient was detected from six CTC-positive patients, and the positive expression of EGFRVIII was also found in the tumor tissue of this patient. CONCLUSIONS: In vivo detection of CTCs had high sensitivity in SCCHN, which might improve CTC application in clinic.

6.
Medicine (Baltimore) ; 95(33): e4339, 2016 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-27537558

RESUMO

Acceptance of chronic pain has increasingly become a significant issue in the field of pain management. Many researchers have suggested that patients with better acceptance of pain are more likely to have better functioning both in physical and psychological status. In many countries, the Chronic Pain Acceptance Questionnaire (CPAQ) and CPAQ-8 have been validated and utilized frequently to measure the pain acceptance of patients with chronic pain. However, the CPAQ and CPAQ-8 yet have not been introduced and validated in Mainland China.In this study, we aimed to translate the English version of the CPAQ into simplified Chinese, make proper cross-cultural adaptations, and validate the psychometric properties of the Chinese version of the CPAQ and the CPAQ-8.The English version of the CPAQ was first linguistically translated and cross-culturally adapted to formulate a Chinese version. Then, we recruited 224 patients from a pain clinic and every participant was asked to finish a series of questionnaires. Finally, statistical analysis was performed to test the psychometric properties of the CPAQ and the CPAQ-8.Both confirmatory factor analysis (CFA) and principal component analysis (PCA) confirmed a 2-factor structure for the CPAQ and the CPAQ-8. Nine out of 10 of the hypotheses were validated for construct validity. The overall intraclass correlation coefficient (ICC) value for the CPAQ and CPAQ-8 were 0.92 and 0.89, respectively. In addition, the Cronbach α values for both the CPAQ and the CPAQ-8 showed excellent test-retest reliability.In conclusion, the original CPAQ was successfully developed into the Chinese version of the CPAQ and CPAQ-8 with excellent validity and reliability. The scores of the CPAQ or CPAQ-8 might be a strong predictor for the physical and psychological function of chronic pain patients. In addition, to improve the satisfaction of surgery patients, we recommend measuring patients' pain acceptance using the CPAQ or CPAQ-8 before and after the surgery. For patients with lower acceptance, psychological interventions may be more effective than treatment that simply reduces symptoms. Finally, we suggest that the Chinese version of the CPAQ and CPAQ-8 are appropriate for use in clinical settings or fundamental research in Mainland China.


Assuntos
Dor Crônica/diagnóstico , Atitude Frente a Saúde , China , Dor Crônica/psicologia , Análise Fatorial , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Manejo da Dor/psicologia , Medição da Dor , Análise de Componente Principal , Reprodutibilidade dos Testes , Inquéritos e Questionários , Tradução
7.
Chin J Nat Med ; 14(1): 48-55, 2016 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-26850346

RESUMO

Insulin resistance is the pathophysiological basis of many diseases. Overcoming early insulin resistance highly significant in prevention diabetes, non-alcoholic fatty liver, and atherosclerosis. The present study aimed at evaluating the therapeutic effects of baicalin on insulin resistance and skeletal muscle ectopic fat storage in high fat diet-induced mice, and exploring the potential molecular mechanisms. Insulin resistance in mice was induced with a high fat diet for 16 weeks. Animals were then treated with three different doses of baicalin (100, 200, and 400 mg·kg(-1)·d(-1)) for 14 weeks. Fasting blood glucose, fasting serum insulin, glucose tolerance test (GTT), insulin tolerance test (ITT), and skeletal muscle lipid deposition were measured. Additionally, the AMP-activated protein kinase/acetyl-CoA carboxylase and protein kinase B/Glycogen synthase kinase 3 beta pathways in skeletal muscle were further evaluated. Baicalin significantly reduced the levels of fasting blood glucose and fasting serum insulin and attenuated high fat diet induced glucose tolerance and insulin tolerance. Moreover, insulin resistance was significantly reversed. Pathological analysis revealed baicalin dose-dependently decreased the degree of the ectopic fat storage in skeletal muscle. The properties of baicalin were mediated, at least in part, by inhibition of the AMPK/ACC pathway, a key regulator of de novo lipogenesis and activation of the Akt/GSK-3ß pathway, a key regulator of Glycogen synthesis. These data suggest that baicalin, at dose up to 400 mg·kg(-1)·d(-1), is safe and able to attenuate insulin resistance and skeletal muscle ectopic fat storage, through modulating the skeletal muscle AMPK/ACC pathway and Akt/GSK-3ß pathway.


Assuntos
Tecido Adiposo/metabolismo , Dieta Hiperlipídica , Flavonoides/farmacologia , Glicogênio Sintase Quinase 3 beta/fisiologia , Resistência à Insulina , Músculo Esquelético/metabolismo , Proteínas Proto-Oncogênicas c-akt/fisiologia , Transdução de Sinais/fisiologia , Proteínas Quinases Ativadas por AMP/metabolismo , Acetil-CoA Carboxilase/metabolismo , Animais , Masculino , Camundongos , Camundongos Endogâmicos C57BL
8.
Artigo em Chinês | MEDLINE | ID: mdl-25989679

RESUMO

OBJECTIVE: To investigate the possibility of thyroidectomy/lobectomy without drainage after surgery. METHOD: Eighty-eight consecutive cases with thyroid mass have been underwent operations including: lobectomy group(42 cases), thyroid lobectomy with contralateral partial thyroidectomy group (17 cases) and total thyroidectomy group (29 cases) from 2013. 06 to 2014. 06 in Nanjing Tongren Hospital. FIfteen patients with thyroid operation in other hospital were collected from 2014. 01 to 2014. 06, the recovery of postoperative incision were compared. RESULTS: Eighty-four cases were smoothly discharged from hospital except 4 thyroidectomy cases suffered from a small amount of effusion in surgical cavity. Our postoperative wound recovery were more in line with the principle of cosmetology compared with other hospital operation group. CONCLUSION: Drainage following thyroidectomy is not essential, if the thyroid surgery is meticulous in each step of the surgery, and can increase the effect of beauty.


Assuntos
Drenagem/métodos , Tireoidectomia , Humanos , Período Pós-Operatório , Doenças da Glândula Tireoide
9.
Cell Physiol Biochem ; 35(6): 2349-59, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-25896320

RESUMO

BACKGROUND/AIMS: Obesity-associated fatty liver disease affects millions of individuals. This study aimed to evaluate the therapeutic effects of baicalin to treat obesity and fatty liver in high fat diet-induced obese mice, and to study the potential molecular mechanisms. METHODS: High fat diet-induced obese animals were treated with different doses of baicalin (100, 200 and 400 mg/kg/d). Whole body, fat pad and liver were weighed. Hyperlipidemia, liver steatosis, liver function, and hepatic Ca(2+)/CaM-dependent protein kinase kinase ß (CaMKKß) / AMP-activated protein kinase (AMPK) / acetyl-CoA carboxylase (ACC) were further evaluated. RESULTS: Baicalin significantly decreased liver, epididymal fat and body weights in high fat diet-fed mice, which were associated with decreased serum levels of triglycerides, total cholesterol, LDL, alanine transaminase and aspartate transaminase, but increased serum HDL level. Pathological analysis revealed baicalin dose-dependently decreased the degree of hepatic steatosis, with predominantly diminished macrovesicular steatosis at lower dose but both macrovesicular and microvesicular steatoses at higher dose of baicalin. Baicalin dose-dependently inhibited hepatic CaMKKß/AMPK/ACC pathway. CONCLUSION: These data suggest that baicalin up to 400 mg/kg/d is safe and able to decrease the degree of obesity and fatty liver diseases. Hepatic CaMKKß/AMPK/ACC pathway may mediate the therapeutic effects of baicalin in high fat diet animal model.


Assuntos
Proteínas Quinases Ativadas por AMP/metabolismo , Acetil-CoA Carboxilase/metabolismo , Quinase da Proteína Quinase Dependente de Cálcio-Calmodulina/metabolismo , Dieta Hiperlipídica/efeitos adversos , Fígado Gorduroso/tratamento farmacológico , Flavonoides/farmacologia , Obesidade/tratamento farmacológico , Tecido Adiposo/efeitos dos fármacos , Tecido Adiposo/metabolismo , Alanina Transaminase/sangue , Animais , Aspartato Aminotransferases/sangue , Peso Corporal/efeitos dos fármacos , Fígado Gorduroso/sangue , Fígado Gorduroso/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Obesos , Obesidade/sangue , Obesidade/metabolismo , Transdução de Sinais/efeitos dos fármacos , Triglicerídeos/sangue
10.
Artigo em Chinês | MEDLINE | ID: mdl-25764924

RESUMO

OBJECTIVE: Thyroid lobectomy can be programmed to operate as "step by step". Each step must be performed meticulously with quality control, so as to minimize the complications of surgery. METHODS: From May 2013 to Auguest 2014, the meticulous thyroid operation was conducted in 72 cases (144 lobectomies). Pre- and post-operative evaluations by strobe laryngoscopy, parathyroid hormone (PTH), and blood calcium examinations were conducted in all cases. RESULTS: Four cases (5.6%) had transient recurrent laryngeal nerve paralysis after surgery, but no permanent recurrent laryngeal nerve paralysis. There was no transient or permanent superior laryngeal nerve paralysis. Transient hypoparathyroidism occurred in 10 cases, with no permanent hypoparathyroidism. No case presented with postoperative bleeding or infection. CONCLUSIONS: "Meticulous operation of thyroid lobectomy" may minimize effectively the complication of surgery.


Assuntos
Glândula Tireoide/cirurgia , Tireoidectomia , Humanos , Hipoparatireoidismo , Nervos Laríngeos , Laringoscopia , Glândulas Paratireoides , Hormônio Paratireóideo , Hemorragia Pós-Operatória , Período Pós-Operatório , Paralisia das Pregas Vocais
11.
Artigo em Chinês | MEDLINE | ID: mdl-25598365

RESUMO

OBJECTIVE: To investigate the protection of the parathyroid in the total thyroidectomy to avoid the postoperative permanent hypoparathyroidism. METHODS: Forty-three consecutive cases underwent total thyroidectomy from June 2013 to June 2014 in Nanjing Tongren Hospital were reviewed. Of them 26 cases with malignant and 17 cases with benign thyroid diseases. RESULTS: Intraoperatively, all 4 parathyroid glands were identified in 27 cases, 3 parathyroids in 10 cases and 2 parathyroids in 6 cases. Intraoperative parathyroid transplantation was performed in 6 cases, including 5 cases with 1 parathyroid transplantation and 1 case with 2 parathyroid transplantation. With the follow-up of 1-3 months after surgery, 10 cases presented with transient hypocalcemia, 8 cases with temporary hypoparathyroidism and no case with permanent hypoparathyroidism. CONCLUSION: Accurate identification and conservation in situ or auto-transplantation in total thyroidectomy could be effective for prevention of postoperative permanent hypoparathyroidism.


Assuntos
Hipoparatireoidismo/cirurgia , Tireoidectomia/métodos , Pesquisa Biomédica , Humanos , Glândulas Paratireoides , Período Pós-Operatório , Doenças da Glândula Tireoide , Transplante Autólogo
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