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Pyroptosis is a novel process of programmed inflammatory necrosis, which is closely related to microbial infections and autoimmune diseases. In recent years it has been proved that Gasdermin D is an executive protein of pyroptosis. Cleaved Gasdermin D leads to the formation of pores in cell membrane to induce pyroptosis, and the release of inflammatory factors such as IL-1β and IL-18 aggravates the occurrence of inflammatory responses. With the deepening of Gasdermin D research, it is gradually clear its protein structure and activation sites, which has other activation pathways apart from inducing inflammasome activation. Activation of Gasdermin D does not necessarily mean cell death, even if it could induce the formation of pores in cell membrane and regulation of inflammatory cytokine secretion. These findings offer insight into Gasdermin D function and change our understanding of pyroptosis and programmed necrosis. The recent progress of Gasdermin D is highlighted in this review.
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The current study aimed to establish simple and quick quality evaluation method of Chishao based on QAMS. Oxypaeoniflorin is used as a marker in the Chishao root. Based on it, the content of other components could be obtained by establishing the mathematical relationship. UPLC method was used to collect data, and the detection wavelengths were 230nm (benzoic acid, paeoniflorin), 263nm (hydroxy paeoniflorin) 274nm (gallic acid, paeoniflorin, catechin), respectively. The stationary phase was an Agilent ZORBAX SB-C18 and the mobile phase was acetonitrile -0.1% formic acid-water. The gradient elution method was adopted at the certain flow rate (0.3 mL/min). The column temperature set 40oC, and the injection volume was 1µL. Multiple reaction monitoring mode was selected for data collection. The linear ranges of benzoic acid, paeoniflorin, hydroxy-paeoniflorin, gallic acid, catechin and paeoniflorinhad good linearity (r ≥0.9995). The UPLC method was established to determine the content of paeoniflorin, benzoic acid, catechin, gallic acid, paeoniflorin, andhydroxy-paeoniflorin in Radix Paeoniae Rubra. In the current study, the method for the chemical components in Radix Paeoniae Rubra to provide the evaluation basis of medicinal effects.
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Catequina , Medicamentos de Ervas Chinesas , Paeonia , Ácido Benzoico , Hidrocarbonetos Aromáticos com Pontes , Cromatografia Líquida de Alta Pressão/métodos , Ácido Gálico , Monoterpenos , Paeonia/química , CaramujosRESUMO
ObjectiveTo explore the intervention effect of Erxian decoction on intestinal microflora after ovariectomy in rats by 16S rRNA gene sequencing. MethodThirty-two female healthy SD rats were randomly divided into a Sham operation (Sham) group, a model (OVX) group, an estrogen (E) group, and an Erxian decoction (EXD) group, with 8 rats in each group. The rats in the E group and the EXD group received 1.8×10-4 g·kg-1 estradiol valerate solution and 9 g·kg-1 Erxian decoction, respectively, and those in the Sham group and the OVX group received an equal volume of distilled water once a day for 16 weeks. After 16 weeks, the levels of serum estrogen and blood lipid were detected. The fecal DNA was extracted, followed by 16S rRNA gene sequencing and analysis. ResultCompared with the Sham group, the OVX group showed reduced serum estrogen level (P<0.01) and increased serum levels of total cholesterol (TC) and low-density lipoprotein cholesterol (LDL-C) (P<0.05). Compared with the OVX group, the E group and the EXD group showed increased serum estrogen level (P<0.01) and reduced TC and LDL-C (P<0.05). Alpha diversity showed that there was no significant change in intestinal microflora diversity after ovariectomy. Beta diversity showed that there were significant differences in the structure of intestinal microflora in the four groups. The intervention of Erxian decoction could improve the changes in intestinal microflora after ovariectomy. LEfSe was used to analyze the differential flora in the four groups. The results showed that the Sham group and the OVX group had 3 differential bacterial phyla and 18 differential bacterial genera, the OVX group and the E group had 1 differential bacterial phylum and 12 differential bacterial genera, and the OVX group and the EXD group had 3 differential bacterial phyla and 5 differential bacterial genera. Estrogen intervention could reverse the change trend of Ruminococcus 1, Anaerovibrio, and Turicibacter in the OVX group. Erxian decoction intervention could reverse the change trend of Bacteroidetes, Firmicutes, Prevotella 9, Ruminococcaceae UCG-014, Ruminococcus 1, and Fusicatenibacter in the OVX group. ConclusionThe structure and function of intestinal microflora in ovariectomized rats changed obviously, and Erxian decoction could ameliorate the change.
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Evodiamine(EVO)is an indoloquinazoline alkaloid isolated from the fruits of Euodia rutaecarpa,which has long been used in traditional Chinese medicine to treat various diseases.Modern medical research shows that evodiamine has various pharmacological activities and extremely high medicinal value such as anti-inflammation,anti-tumor,losing weight,treating Alzheimer's,and antibacterium.Evodiamine can interact with a variety of proteins.For example,the interaction between evodiamine and TRPV1 induces its activation to exert anti-inflammatory and losing weight; the interaction with DNA topoisomerase I inhibits its function and thus inhibit bacterial proliferation; the interaction with tubulin promotes NLRP3 inflammasome activation,and exerts anti-tumor and anti-inflammatory functions.In recent years,with the in-depth study of evodiamine,pharmacological research and mechanism of evodiamine has significant improvement.Recent progress of pharmacological effect of evodiamine is highlighted in this review.
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Objective:To observe the changes of myocardial microvessel density, microvascular endothelial cell morphology and hemorheology in ovariectomized rats and explore the interventional effects of Erxian decoction. Method:Thirty-two healthy 10 week-old female SPF SD rats were randomly divided into sham operation group, model group, estrogen group (estradiol valerate, 0.18 mg·kg-1) and Erxian decoction group (9 g·kg-1). The rats were intragastrically administered 2 weeks after ovariectomy, once a day for 16 weeks. Sham operation groups and model groups were given equal volumes of purified water. After 16 weeks of administration, the cardiac function was measured by noninvasive ultrasound cardiogram (UCG), CD34 in the myocardial tissue was tested by immunofluorescence staining to measure the microvessel density, the morphological structure of microvessels of myocardial tissue were detected by transmission electron microscope, the levels of estrogen (E2) in rat plasma were detected by radioimmunoassay, the levels of endothelin-1 (ET-1), prostacyclin I2 (PGI2), thromboxane A2 (TXA2), endothelial growth factor (VEGF), and von Willebrand Factor (vWF) in rat plasma were detected by enzyme-linked immuno sorbent assay (ELISA), four items of coagulation was detected by blood coagulation analyzer, whole blood viscosity and plasma viscosity were detected by hemorheology. Result:Compared with sham operation group, the ejection fraction (EF) decreased (P<0.01), the left ventricular short axis shortening rate (FS) decreased (P<0.01), and the left ventricular end systolic volume (LVVols) increased (P<0.01), myocardial microvessel density significantly reduced (P<0.01), the endothelial cells were swollen and the cytoplasm was cavitation, E2 in rat plasma decreased (P<0.01), ET-1, VEGF, vWF increased (P<0.01), prostacyclin I2 /thromboxane A2 (PGI2/TXA2) decreased (P<0.01), plasma activated partial prothrombin time (APTT) decreased (P<0.01), fibrinogen (FIB) increased (P<0.01), whole blood viscosity, plasma viscosity, and cassone viscosity increased (P<0.01), whole blood high-cut, low-cut index, and red blood cell (RBC) aggregation index increased (P<0.05) in model group. Compared with model group, EF and FS increased (P<0.05), LVVols decreased (P<0.05), myocardial microvessel density significantly increased (P<0.01), the endothelial cell edema was improved, and transport vesicles were clearly visible, E2 in rat plasma increased (P<0.01), ET-1, VEGF, decreased (P<0.01), PGI2/TXA2 increased (P<0.01), APTT increased (P<0.01), whole blood viscosity, whole blood high shear relative index, RBC aggregation index decreased (P<0.05), Kasson viscosity and plasma viscosity decreased (P<0.01) in Erxian decoction group. Conclusion:Erxian decoction increases myocardial microvessel density, protects the structural integrity of microvascular endothelial cells, improves its endothelial secretion function and hemorheology in ovariectomized rats, and protects heart function.
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Objective::To investigate the evolution of cardiac function and blood pressure in ovariectomized rats and the effect and mechanism of Erxiantang. Method::Healthy 10-week-old female SPF SD rats were randomly divided into sham operation group, model group, estrogen group(estradiol valerate, 0.18 mg·kg-1) and Erxiantang group(7.5 g·kg-1). The rats were intragastrically administered 2 weeks after ovariectomy, once a day for 12 weeks.Sham operation groups and model groups were given equal volumes of purified water.At the 4th week, 8th week, and 12th week after administration, the cardiac function, blood pressure, and levels of estrogen (E2) in rat serum were measured by non-invasive ultrasound cardiogram (UCG), tail artery detection techniques and radioimmunoassay.The levels of endothelin-1 (ET-1) and angiotensin 2(Ang Ⅱ) in rat serum were detected by enzyme-linked immuno sorbent assay (ELISA). The cardiac morphology and apoptosis were detected by hematoxylin-eosin (HE) staining, electron microscopy and Terminal-deoxynucleoitidyl transferase mediated nick end labeling (TUNEL). Result::Compared with sham operation group, the ejection fraction (EF) decreased and the left ventricular end systolic volume (LVVols) increased in the model group at 4th week after administration(P<0.05). There was no significant difference in cardiac function between the groups at 8th week.The left ventricular end diastolic diameter (LVIDs), LVVols, left ventricular end diastolic diameter (LVIDd), and left ventricular end diastolic volume (LVVold) were significantly increased in the model group at 12th week (P<0.01). At the 4th weeks, 8th week and 12th week, the systolic blood pressure (SBP) of the model group increased (P<0.05) and showed an increasing trend, and the diastolic blood pressure (DBP) did not change significantly.At the 12th week, the levels of E2 in serum decreased (P<0.05), ET-1 and Ang Ⅱ increased of the model group (P<0.01). The cardiac myofibrils were irregular, some myofilament was broken, and mitochondrial palsy was disordered, broken or disappeared, and cardiac apoptosis increased (P<0.01). Compared with the model group, myocardial contraction and diastolic function were significantly improved in Erxian decoction group, and blood pressure was decreased.The levels of E2 in serum was increased (P<0.05). The levels of ET-1 was decreased (P<0.05), and AngⅡ in serum was significantly decreased (P<0.01). The mitochondrial morphological structure was improved and the cardiac apoptotic rate was significantly decreased (P<0.01). Conclusion::After the ovariectomy, the rats showed a series of pathological changes such as decreased heart function and increased blood pressure.Compared with the decrease of heart function, the changes of blood pressure appeared earlier.Erxiantang exerts its intervention on cardiac function and blood pressure in ovariectomized rats by regulating E2, blood active substances and cardiac apoptosis.
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Objective: To observe the clinical efficacy of addition and subtraction therapy of Taohong Siwutang combined with Chaihu Shugansan to unstable angina pectoris with type A behavior pattern (Qi stagnation and blood stasis syndrome), and investigate its effects on proinflammatory factors and serotonin (5-HT). Method: One hundred twenty-four patients were randomly divided into control group (60 cases) and observation group(64 cases) by random number table. Patients in control group got Aspirin enteric-coated tablets, 100 mg/time, 1 time/day. Tigrillo tablets, 90 mg/time, 2 times/days. Metoprolol tartrate tablets, 50 mg/time, 2 times/days. Simvastatin tablets, 10 mg/time, 1 time/day. Nitroglycerin tablets, 0.5 mg/time. Based on the treatment in control group, patients in observation group also received addition and subtraction therapy of Taohong Siwutang combined with Chaihu Shugansan, 1 dose/day. The treatment course was 8 weeks in both groups. Number of attacks, duration, degree of pain and usage of nitroglycerin were recorded for every week. Before and after treatment, electrocardiogram was also recorded. And levels of triglyceride (TG), total cholesterol (TC), high density lipoprotein cholesterol (HDL-C), low density lipoprotein cholesterol (LDL-C), interleukin-6 (IL-6), tumor necrosis factor-α(TNF-α), hypersensitive C-reactive protein and 5-HT were detected. In addition, scores of Seattle Angina Scale (SAQ) and Qi stagnation and blood stasis syndrome were graded. Result: In the rank sum test, the curative effect in electrocardiogram of observation group was better than that of control group (Z=1.965, PPPPα and 5-HT in observation group were lower than those in control group (PPConclusion: On the basis of conventional western medicine, addition and subtraction therapy of Taohong Siwutang combined with Chaihu Shugansan can further control angina attack, relieve clinical symptoms, improve quality of life, regulate lipid metabolism, and can inhibit expression of proinflammatory factors and 5-HT, so it can play a role in stabilizing the disease.
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Globozoospermia has been reported to be a rare but severe causation of male infertility, which results from the failure of acrosome biogenesis and sperm head shaping. Variants of dpy-19-like 2 (DPY19L2) are highly related to globozoospermia, but related investigations have been mainly performed in patients from Western countries. Here, we performed a screening of DPY19L2 variants in a cohort of Chinese globozoospermic patients and found that five of nine patients carried DPY19L2 deletions and the other four patients contained novel DPY19L2 point mutations, as revealed by whole-exome sequencing. Patient 3 (P3) contained a heterozygous variant (c.2126+5G>A), P6 contained a homozygous nonsense mutation (c.1720C>T, p.Arg574*), P8 contained compound heterozygous variants (c.1182-1184delATC, p.Leu394_Ser395delinsPhe; c.368A>T, p.His123Arg), and P9 contained a heterozygous variant (c.1182-1184delATCTT, frameshift). We also reported intracytoplasmic sperm injection (ICSI) outcomes in the related patients, finding that ICSI followed by assisted oocyte activation (AOA) with calcium ionophore achieved high rates of live births. In summary, the infertility of these patients results from DPY19L2 dysfunction and can be treated by ICSI together with AOA.
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Adulto , Feminino , Humanos , Masculino , Gravidez , Acrossomo , China , Códon sem Sentido , Proteínas de Membrana/genética , Mutação Puntual , Resultado da Gravidez , Taxa de Gravidez , Deleção de Sequência , Cabeça do Espermatozoide , Injeções de Esperma Intracitoplásmicas , Teratozoospermia/genética , Sequenciamento do ExomaRESUMO
Based on the outpatient interview and literature review,the initial framework of the outpatient experience of human caring scale was formed with 9 dimensions of outpatient process.The research aim was to improve the scale by Delphi method.Sixteen experts in medical management,human caring or medical education were invited to evaluate the importance of the dimensions and items of the scale and provided some expertise via filling out the Delphi consultation questionnaires twice in the consulting round.In the first round,the recovery rate showing the experts' positivity was 80%;the coefficient of reliability (Cr) ascertaining the authority of the evaluation was 0.92;the mean and full mark ratios responding the concentration of the evaluation were 2.88-4.94 and 6.25%-93.75% respectively;the coefficients of variation (CV) and the Kendall's W determining the concordance of the evaluation were 5.06%-52.15% and 0.21-0.24 respectively.In the second round,the recovery rate was 93.75%;the Cr was 0.93;the mean was 3.93-4.93;the full mark ratios were 26.67%-93.33%;the Kendall's W was 0.14-0.31,the CV was 5.25%-23.61%.Via the two-round Delphi study,the scale that included 10 dimensions and 61 items has been improved.Ten dimensions are pre-hospital medical service,guidance,registration,waiting,diagnosis & treatment,paying,inspection & assay,medicine receiving,therapy/injection/transfusion and global evaluation.It was concluded that Chinese scholars have paid high attention to human caring and outpatient experience.The experts have given high agreements about the dimensions which were established with Chinese outpatient process.The dimensions are different from the similar researches about outpatient experience study.In the future,it is necessary to survey the outpatients to test the construct validity,internal consistency reliability and others of the scale to improve the scale.
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Based on the outpatient interview and literature review,the initial framework of the outpatient experience of human caring scale was formed with 9 dimensions of outpatient process.The research aim was to improve the scale by Delphi method.Sixteen experts in medical management,human caring or medical education were invited to evaluate the importance of the dimensions and items of the scale and provided some expertise via filling out the Delphi consultation questionnaires twice in the consulting round.In the first round,the recovery rate showing the experts' positivity was 80%;the coefficient of reliability (Cr) ascertaining the authority of the evaluation was 0.92;the mean and full mark ratios responding the concentration of the evaluation were 2.88-4.94 and 6.25%-93.75% respectively;the coefficients of variation (CV) and the Kendall's W determining the concordance of the evaluation were 5.06%-52.15% and 0.21-0.24 respectively.In the second round,the recovery rate was 93.75%;the Cr was 0.93;the mean was 3.93-4.93;the full mark ratios were 26.67%-93.33%;the Kendall's W was 0.14-0.31,the CV was 5.25%-23.61%.Via the two-round Delphi study,the scale that included 10 dimensions and 61 items has been improved.Ten dimensions are pre-hospital medical service,guidance,registration,waiting,diagnosis & treatment,paying,inspection & assay,medicine receiving,therapy/injection/transfusion and global evaluation.It was concluded that Chinese scholars have paid high attention to human caring and outpatient experience.The experts have given high agreements about the dimensions which were established with Chinese outpatient process.The dimensions are different from the similar researches about outpatient experience study.In the future,it is necessary to survey the outpatients to test the construct validity,internal consistency reliability and others of the scale to improve the scale.
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In this study,we estimated the application value of detecting Mycobacterium tuberculosis (MTB) specific IgG/IgM antibodies for tuberculosis diagnosis with colloidal gold immunochromatography assay (GICA).We collected 332 effective serum samples and their background information,including 260 patients with tuberculosis and 72 healthy individuals.The means of GICA was used to detect MTB specific IgG/IgM antibodies.Results were compared with the clinical diagnosis and the results of bacteriological tests.The SPSS 22.0 software was used to analyze the results,and when P<0.05 the difference was statistically significant.The sensitivity and specificity of GICA were 41.15% and 91.67%,and the sensitivity of the bacterial positive and negative patients were 51.38% and 33.77%,respectively.The positive rate of IgG/IgM antibodies detection with GICA (41.15%) was much higher than that of bacteria with acid-fast stain of sputum smear (18.84%) and sputum bacteria cultivation (36.15 %) (P < 0.05) respectively.The positive rate of the combination of tuberculosis antibody detection,sputum bacterial culture and sputum smear was 61.54%,higher than the result of single method or combination of two methods.The detection of specific antibodies against MTB in serum with GICA is sensitive,specific,rapid and convenient,which can be used in clinical screening.Meanwhile,there are still certain limitations of this method,and the sensitivity and specificity need to be improved.Therefore,the GICA can be used as an auxiliary diagnosis combined with sputum bacteriology,imaging test and clinical features rather than diagnose tuberculosis alone.
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Chinese medicine prescription is the main form and means to treat diseases in traditional Chinese medicine (TCM). However, the disjointing situation is generally present between "systemic" and "local", "macro" and "micro", " process" and "activity evaluation" in the study of TCM at present. An urgent task for the modernization of TCM is to establish new strategies and methods which can reflect the overall characteristics of TCM. The introduction of integrative pharmacology provided a feasible approach to solve the problem of the fragmentation of TCM. Internet-based computation platform method was adopted in this study to explore the active molecular mechanism of Yinchenhao decoction in the treatment of cirrhosis. Based on the analysis of the functional integration of Internet-based Computation Platform V1.0 version software, the "core components-key target- main pathway" multidimensional network of Yinchenhao decoction in treatment of cirrhosis disease was constructed to explore the potential molecular mechanism of Yinchenhao decoction in treatment of cirrhosis from multiple perspectives. The molecular mechanism analysis of Yinchenhao decoction showed that Yinchenhao decoction can achieve the therapeutic effect on cirrhosis and the mechanism might be associated with oxidative phosphorylation, energy metabolism, circulatory system, glycerophospholipid metabolis, lipid metabolism and other pathways. Yinchenhao decoction in treatment of cirrhosis may be associated with energy metabolism and lipid metabolism.
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At present, the disjointing situation is generally present between "systemic"and "local", "macro" and "micro", " process" and "activity evaluation" in the study of traditional Chinese medicine (TCM). An urgent task for the modernization of TCM is to establish new strategies and methods which can reflect the overall characteristics of TCM. The introduction of integrative pharmacology provided a feasible approach to solve the problem of the fragmentation of TCM. Internet-based computation platform method was adopted in this study to explore the active molecular mechanism of Mylabris in the treatment of colorectal cancer. Based on the analysis of the functional integration of internet-based computation platform V1.0 version software, the "core components-key target- main pathway" multidimensional network of Mylabris in treatment of colorectal cancer disease was constructed to explore the potential molecular mechanism of Mylabris in treatment of colorectal cancer from multiple perspectives. The results showed that Mylabris can treat the colorectal cancer and the mechanism might be associated with amino acid metabolism, NF-κB signaling pathway, immune system, endocrine system, nervous system, and chemokine signal transduction pathway, epithelial cell signaling in helicobacter pylori infection, T cell receptor signaling pathway, B cell receptor signaling pathway and so on.
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<p><b>OBJECTIVE</b>To identify potential serum biomarkers for distinguishing between latent tuberculosis infection (LTBI) and active tuberculosis (TB).</p><p><b>METHODS</b>A proteome microarray containing 4,262 antigens was used for screening serum biomarkers of 40 serum samples from patients with LTBI and active TB at the systems level. The interaction network and functional classification of differentially expressed antigens were analyzed using STRING 10.0 and the TB database, respectively. Enzyme-linked immunosorbent assays (ELISA) were used to validate candidate antigens further using 279 samples. The diagnostic performances of candidate antigens were evaluated by receiver operating characteristic curve (ROC) analysis. Both antigen combination and logistic regression analysis were used to improve diagnostic ability.</p><p><b>RESULTS</b>Microarray results showed that levels of 152 Mycobacterium tuberculosis (Mtb)-antigen- specific IgG were significantly higher in active TB patients than in LTBI patients (P < 0.05), and these differentially expressed antigens showed stronger associations with each other and were involved in various biological processes. Eleven candidate antigens were further validated using ELISA and showed consistent results in microarray analysis. ROC analysis showed that antigens Rv2031c, Rv1408, and Rv2421c had higher areas under the curve (AUCs) of 0.8520, 0.8152, and 0.7970, respectively. In addition, both antigen combination and logistic regression analysis improved the diagnostic ability.</p><p><b>CONCLUSION</b>Several antigens have the potential to serve as serum biomarkers for discrimination between LTBI and active TB.</p>
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Adolescente , Adulto , Idoso , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Adulto Jovem , Anticorpos Antibacterianos , Especificidade de Anticorpos , Antígenos de Bactérias , Biomarcadores , Sangue , Tuberculose Latente , Sangue , Diagnóstico , Modelos Logísticos , Mycobacterium tuberculosis , Análise Serial de Proteínas , Métodos , Proteoma , Genética , Proteômica , Métodos , Curva ROCRESUMO
Objective To investigate the effect of rosiglitazone on the proliferation and apoptosis of colon cancer cells and the changes in the AKT/GSK3β signaling pathway. Methods Different concentrations of rosiglitazone ( 20. 0μmol/L, 40. 0 μmol/L, 80. 0 μmol/L) were used to treat colon cancer HT29 cells and HCT116 cells. Cell proliferation was detected by MTT assay. Annexin V FITC/PI cell death detection kit was used to test the cell apoptosis rate. The expression of apoptotic protein Bcl-2, Bax and Akt, GSK3β were detected by Western blot. Results Different concentrations of rosiglitazone had different effect on the proliferation of colon cancer cells compared with the blank control group, and showed a dose dependence (P< 0. 01). With the increase of rosiglitazone dose, the apoptosis-inducing effect @was increased dose-dependently (P< 0. 01). When the cells were treated with rosiglitazone for 48 h, the expressions of Bcl-2/Bax, p-GSK3β, and p-Akt were significantly decreased compared with the blank control group (P< 0. 01), but the expression level of Akt and GSK3β was not significantly different compared with the blank control group ( P > 0. 05 ) . Conclusions Rosiglitazone significantly induces apoptosis and inhibits the proliferation of HT29 cells. It may be via inhibiting Akt/GSK3β signaling pathway and change the ratio of Bcl-2/Bax.
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BACKGROUND: Momordica charantia (MC) is an edible medicinal plant that is known for its diversified biological functions. Momordica Antiviral Protein 30kD (MAP30) is a type I single chain ribosome-inactivating protein (RIP) isolated from the mature fruit and seeds of MC. Since MAP30 content in MC is limited, the study aim was to generate the recombinant MAP30 protein using prokaryotic expression system and determine its apoptotic/growth inhibitory effects on bladder cancer 5637 cells. METHODS: MAP30 gene was amplified by PCR from MC genomic DNA and identified by sequencing. The target gene was inserted into pET-28a (+) vector and transformed into E. coli BL21 (DE3) cells. Positive clones were selected by PCR. Recombinant protein was efficiently expressed under induction with 1.0 mM Isopropylthio-ß-D-galactoside (IPTG) at 30° C for 4 hours. Cytotoxicity studies were performed using MTT assay by treating 5637 bladder cancer cells with 100 µg/mL, 200 µg/mL, and 400 µg/mL concentrations of MAP30 for 24 hours and 48 hours, respectively. Flow cytometry was used to measure the apoptosis of MAP30-treatedcells in time course experiments. RESULTS: Full-length MAP30 gene was successfully expressed in Escherichia coli (E. coli) BL21 strain and MAP30 recombinant protein inhibited the growth of bladder cancer 5637 cells at 200 µg/mL and 400 µg/mL concentrations by inducing apoptosis of target cells in a dose- and time-dependent manner. CONCLUSION: It was, therefore, concluded that the MAP30 recombinant protein displayed potent antitumor activity in vitro.
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Momordica charantia/metabolismo , Proteínas Inativadoras de Ribossomos Tipo 2/biossíntese , Proteínas Inativadoras de Ribossomos Tipo 2/uso terapêutico , Neoplasias da Bexiga Urinária/tratamento farmacológico , Apoptose/efeitos dos fármacos , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Ensaios de Seleção de Medicamentos Antitumorais , Escherichia coli/metabolismo , Humanos , Proteínas Recombinantes/biossínteseRESUMO
Interleukin 8 (IL8) is an important chemokine that elicits host immune response against tuberculosis (TB). However, whether there is an association between IL8 gene polymorphism and TB susceptibility in the Chinese population is unknown. IL8 gene was amplified and sequenced to search for nucleotide polymorphisms among the Chinese population. Four single nucleotide polymorphisms (SNPs) were identified, selected, and analyzed in a cohort of 438 patients with TB and 536 healthy controls. Allelic, genotypic, and haplotypic analysis demonstrated that the distribution of the four IL8 SNPs between patients with TB and healthy controls was not significantly different (P>0.05). The four IL8 SNPs detected in this study were not associated with TB susceptibility in the Chinese population. Secretion of IL8 by peripheral blood cells was greatly stimulated upon exposure to Mycobacterium tuberculosis whole cell extract, but such enhanced secretion was not associated with the IL8 rs4073 alleles.
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Humanos , Povo Asiático , Genética , China , Predisposição Genética para Doença , Interleucina-8 , Genética , Polimorfismo de Nucleotídeo Único , Tuberculose , GenéticaRESUMO
We evaluate the performance of Xpert MTB/RIF for the diagnosis of extrapulmonary tuberculosis (EPTB) in China. The performance of Xpert was evaluated compared to the composite reference standard (CRS), drug susceptibility testing (DST), and imaging examination. The overall sensitivity and specificity of Xpert were 64.1% (195/304) and 100% (24/24), respectively, using CRS as the gold standard. The sensitivity was significantly higher than that of culture for pus (P<0.05). The proportion of EPTB-positive cases diagnosed by imaging was two times more than that diagnosed using Xpert; however, 6 out of 19 cases may have been overdiagnosed by imaging. Compared to phenotypic DST, the sensitivity and specificity of Xpert were 80% (12/15) and 100% (75/75), respectively. Considering its high sensitivity and specificity, Xpert MTB/RIF may be used as a rapid initial test for EPTB diagnosis, and may also support a quicker decision on the treatment regimen. The combination of imaging and Xpert testing could provide high efficiency and accurate diagnosis of suspected EPTB.
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Humanos , Proteínas de Bactérias , Genética , Metabolismo , China , RNA Polimerases Dirigidas por DNA , Genética , Metabolismo , Testes Diagnósticos de Rotina , Métodos , Farmacorresistência Bacteriana , Testes de Sensibilidade Microbiana , Mycobacterium tuberculosis , Genética , Metabolismo , Estudos Retrospectivos , Rifampina , Farmacologia , Sensibilidade e Especificidade , Escarro , Tuberculose , Tuberculose Pulmonar , Diagnóstico , MicrobiologiaRESUMO
<p><b>OBJECTIVE</b>To re-analyze the data published in order to explore plausible biological pathways that can be used to explain the anti-aging effect of curcumin.</p><p><b>METHODS</b>Microarray data generated from other study aiming to investigate effect of curcumin on extending lifespan of Drosophila melanogaster were further used for pathway prediction analysis. The differentially expressed genes were identified by using GeneSpring GX with a criterion of 3.0-fold change. Two Cytoscape plugins including BisoGenet and molecular complex detection (MCODE) were used to establish the protein-protein interaction (PPI) network based upon differential genes in order to detect highly connected regions. The function annotation clustering tool of Database for Annotation, Visualization and Integrated Discovery (DAVID) was used for pathway analysis.</p><p><b>RESULTS</b>A total of 87 genes expressed differentially in D. melanogaster melanogaster treated with curcumin were identified, among which 50 were up-regulated significantly and 37 were remarkably down-regulated in D. melanogaster melanogaster treated with curcumin. Based upon these differential genes, PPI network was constructed with 1,082 nodes and 2,412 edges. Five highly connected regions in PPI networks were detected by MCODE algorithm, suggesting anti-aging effect of curcumin may be underlined through five different pathways including Notch signaling pathway, basal transcription factors, cell cycle regulation, ribosome, Wnt signaling pathway, and p53 pathway.</p><p><b>CONCLUSION</b>Genes and their associated pathways in D. melanogaster melanogaster treated with anti-aging agent curcumin were identified using PPI network and MCODE algorithm, suggesting that curcumin may be developed as an alternative therapeutic medicine for treating aging-associated diseases.</p>
Assuntos
Animais , Envelhecimento , Genética , Ciclo Celular , Genética , Curcumina , Farmacologia , Proteínas de Drosophila , Genética , Metabolismo , Drosophila melanogaster , Genética , Regulação da Expressão Gênica , Redes Reguladoras de Genes , Genes de Insetos , Biossíntese de Proteínas , Genética , Mapas de Interação de Proteínas , Genética , Receptores Notch , Genética , Metabolismo , Ribossomos , Metabolismo , Transdução de Sinais , Genética , Proteína Supressora de Tumor p53 , Metabolismo , Via de Sinalização Wnt , GenéticaRESUMO
<p><b>BACKGROUND</b>Currently, migration has become one of the risk factors of high burden of tuberculosis in China. This study was to explore the influence of mass migration on the dynamics of Mycobacterium (M.) tuberculosis in Beijing, the capital and an urban area of China.</p><p><b>METHODS</b>Three hundred and thirty-six M. tuberculosis strains from the Changping district, where the problem of urban migrants was more pronounced than in other Beijing regions, were genotyped by Spoligotyping, large sequence polymorphisms (LSPs 105 and 181), and variable number tandem repeat (VNTR) typing. Based on the genotype data, the phylogeny of the isolates was studied.</p><p><b>RESULTS</b>In Changping district, the proportion of Beijing lineage M. tuberculosis isolates amounted to 89.0% (299/336), among which 86.6 % (252) belonged to the modern lineage. The frequency of modern Beijing lineage strains is so high (around 75% (252/336)) that associated risk factors affecting the tuberculosis epidemic cannot be determined. The time to the most recent common ancestor (TMRCA) of the Beijing lineage strains was estimated to be 5073 (95% CI: 4000-6200) years. There was no significant difference in the genetic variation of Beijing isolates from urban migrants and local residents.</p><p><b>CONCLUSIONS</b>The clone of modern Beijing lineage M. tuberculosis, which is dominant in the Beijing area, most likely started to expand with the five thousand-year-old Chinese civilization. In the future, with the urbanization in the whole of China, modern Beijing lineage M. tuberculosis may gain the larger geographical spread.</p>