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OBJECTIVE@#To explore the effect of dichloromethane extraction phase of ethanol extract from stem of Patrinia scabiosaefolia Fisch.(DPSS) on proliferation and differentiation of K562 cells and its related mechanism.@*METHODS@#MTT assay was used to detect the effects of DPSS at 0, 25, 50, 100 and 200 μg/ml on the proliferation of K562 cells at 24, 48 and 72 hours. Flow cytometry was used to analyze the changes of cell cycle and apoptosis at 24 and 48 hours. Wright-Giemsa staining was used to observe the morphological changes of K562 cells. The cell surface antigens CD33 and CD11b were detected by flow cytometry.@*RESULTS@#The proliferation of K562 cells treated with different concentrations of DPSS was inhibited in a time-dose dependent manner (r=-0.96). Cell cycle analysis showed that with the increase of DPSS concentration, cells in G2/M phase increased (r=0.88), and cells were blocked in G2/M phase. Flow cytometry results showed that with the apoptosis rate of K562 cells was the highest when treated with 200 μg/ml DPSS for 48 h. Morphological observation showed that the K562 cell body increased, the amount of cytoplasm increased, the ratio of nucleus to cytoplasm decreased, and the nuclear chromatin was rough after DPSS treatment. Cell differentiation antigen, CD33 and CD11b, were positively expressed after treated with DPSS.@*CONCLUSION@#DPSS can induce apoptosis through cell cycle arrest, inhibit the proliferation of K562 cells, and induce K562 cells to differentiate into monocytes, which has a potential anti-leukemia effect.
Subject(s)
Humans , K562 Cells , Patrinia , Methylene Chloride/pharmacology , Apoptosis , Cell Proliferation , Cell DifferentiationABSTRACT
Objective To investigate the tumor molecular mechanism of Hedgehog/Gli in promoting the epithelial-mesenchymal transition (EMT) in gastric cancer AZ521 cells. Methods After 24 h of treatment with GANT61,the mRNA expression of Gli1,Gli2, N-cadherin,and E-cadherin in the AZ521 cell line were detected by real-time fluorescence quantitative PCR. A Western blotting assay was conducted to determine the expression of the above cytokines,p-AKT and AKT. The effect of GANT61 on invasion was observed by transwell assay. N-Shh stimulation of the Hedgehog pathway was conducted to confirm the changes in these cytokines. Results GANT61 significantly downregulated the mRNA expression of Gli1,Gli2,and N-cadherin,but upregulated E-cadherin mRNA expression. The Western blotting assay revealed that GANT61 downregulated the protein expression of Gli1,Gli2,p-AKT,and N-cadherin,but upregulated E-cadherin expression. Furthermore,GANT61 inhibited the invasion. N-Shh proteins up-regulated Gli1,Gli2,and N-cadherin mRNA,protein expression and p-AKT protein expression,but downregulated E-cadherin mRNA and protein expressions. N-Shh promoted the invasion of tumor cells. Conclusion Downregulation of Gli1 and Gli2 can inhibit the invasion and metastasis in gastric cancer cells,which may be related to the promotion of EMT by Gli through the PI3K/AKT pathway.
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Objective To explore the best intervention based on evidences of enteral nutrition support complicated with diarrhea in critically ill patients and evaluate its clinical effects.Methods Evidence was obtained using Evidence-based Nursing,and was implemented in clinical practice after localization,we formulated numing recommendations for regulating nurses' clinical practice regarding enteral nutrition support complicated with diarrhea in ICU patients;we conducted multiple online and offline training for nurses.The rate of diarrhea in ICU patients undergoing enteral nutrition support before and after using best evidence was compared,and awareness and implementation of best evidence among nurses before and after training were also compared.Results After adapting the best evidence,the rate of enteral nutrition associated diarrhea decreased from 25.58％ to 13.89％;ICU nurses' diarrhea identification and evaluation,analysis of influencing factors of enteral nutrition associated diarrhea,selection of enteral nutrition formula,nasal feeding,etc.,were significantly improved (P＜0.05);there was no significant difference in enteral nutrition infusion,and nutrition preparation and preservation (P＞0.05);rare of implementation of best evidence in ICU nurses was greater than 80％.Conclusion Evidence-based practice can effectively prevent enteral nutrition associated diarrhea in ICU patients.Nurses can solve enteral nutrition associated diarrhea using scientific nursing methods through implementation of best evidence,in order to nursing quality.
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Extracting and encoding rhythmic information is very important for brain functions, as it is the foundation of speech recognition,music appreciation and rhythmic movement et al.However,our knowledge of how the neural system processes rhythm information of external inputs remains limited. In the present paper,we review a neural network model of the scale-free topology can serve as an efficient way to encode rhythm information. In the model, neurons are connected by either electrical synapses or chemical synapses with strengths decreasing with the connectivity of neurons, so that hub neurons are difficult to activate. To encode rhythm information, hub neurons trigger synchronous firing across the network, while loops formed by low-degree neurons determine the rhythm of synchronous firing. This model successfully reproduces the long-period synchronous firing observed in experimental data, and indicates that the neural system can encode temporal information via local dynamics in a distributed way.This review aims to elaborate and summarize the mechanism by which neurons encode rhythmic information.
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Objective To study the inhibitory effects of GANT61, as an inhibitor of Gli, on the growth of human esophageal adenocarcinoma cell lines OE19 and OE33, and their mechanisms thereof. Methods After treating with different concentrations of GANT61(30,20,13.333 3,8.888 8,5.925 9,3.950 6,2.633 7,1.755 8,1.170 5μmol/L),the cell viabilities of OE19 and OE33 were detected by MTS method, which expressed by IC50. The Gli1and Gli2 mRNA expressions treated with GANT61(10 μmol/L GANT61) or DMSO for 24 h were detected in OE19 and OE33 cell lines by real time fluorescence quantitative PCR. The protein expressions of Gli1, Gli2 and CyclinD1 treated with GANT61 or DMSO for 24 h were detected in OE19 and OE33 cell lines by Western blot assay. Transwell invasion assay was performed to evaluate the inhibiting effect on OE19 and OE33 cell invasion by the treatment of GANT61 or DMSO. Results The IC50 of GANT61 was 8.08μmol/L in OE19 and 9.65μmol/L in OE33 cells. Compared with DMSO group, Gli1 and Gli2 mRNA expressions and Gli1,Gli2 and CyclinD1 protein expressions were significantly decreased in OE19 and OE33 cells of GANT61 group (P<0.05). The number of penetrating cells was significantly reduced in OE19 and OE33 cells of GANT61 group compared with that of DMSO group (P<0.01). Conclusion GANT61 can inhibit the growth and invasion of esophageal neoplasms cells by down-regulating Gli1 and Gli2 mRNA expression,which indicates that Hedgehog signaling pathway may play an important role in carcinogenesis and progression of esophageal adenocarcinoma.
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Objective To study the relationship between expression of Shh and Gli1 in Hedgehog(Hh) signal pathway and esophageal squamous cell carcinoma (ESCC).Methods Expressions of Shh andGli1 were detected in 64 cases primary tumor tissues and 24 cases normal esophageal mucosa with SP immunohistochemistal method.Results There were significant differences of Shh and Gli1 expressions between esophageal carcinoma and normal mucosa epithelium (67.19％ (43/64) vs.4.17％ (1/24), 60.94％ (39/64) vs.4.17％ (1/24);x2 =27.729,22.689;P＜0.01).There were significant differences of Shh expression between different degrees of differentiation in esophageal squamous cell carcinoma, metastasis of lymph node and different clinical staging group (x2 =3.873, 11.349,6.429;P ＜ 0.05 or P＜ 0.01).The positive expression rate of Gli1 showed significant differences between different degrees of differentiation in esophageal squamous cell carcinoma, metastasis of lymph node and different clinical staging group (x2=12.598, 9.741,26.341;P ＜0.01).There was a positive relationship between Shh and Gli1 expression(r =0.259, P＜0.05).Conclusion Hh signaling pathway is abnormally activated in ESCC, so that Shh and Gli1 play important roles in carcinogenesis and progression ofesophageal carcinoma.The Hh signaling pathway may be a useful target in ESCC.
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<p><b>OBJECTIVE</b>To establish a comprehensive quality control method for total flavonoid of Fructus Aurantii.</p><p><b>METHODS</b>RP-HPLC and spectrophotometry were applied for the quantitative and fingerprint analysis of total flavonoid of Fructus Aurantii. The contents of naringin and neohesperidin were determined on an Agilent SB-C₁₈column (4.6 mm × 250 mm, 5 μm). The mobile phase was composed of 0.02 % H₃PO₄ and CH₃CN (80:20). The flow rate was 1 ml/min with DAD detected at 280 nm. The column temperature was maintained at 35°C. The fingerprints were developed on an Agilent SB-C₁₈ column (4.6 mm × 250 mm, 5 μm). The mobile phase was composed of 0.5 % HAc and CH₃OH with a linear gradient elution. The ratio of 0.5 % HAc and CH₃OH was: 0 min, 80:20; 10 min, 60:40; 35 min, 30:70; 50 min, 0:100. The flow rate was 1 ml/min with DAD detected at 320 nm. The column temperature was maintained at 30 degree. Meanwhile, the contents of total flavonoid were determined at 283 nm.</p><p><b>RESULT</b>The contents range of naringin, neohesperidin and total flavonoid were 38.3 %- 47.2%, 21.0 %- 28.5% and 79.9%-88.6 %, respectively. The fingerprints of the effective fractions showed 12 common peaks and the fingerprint similarity was all above 98.0 % compared with the standard chromatogram.</p><p><b>CONCLUSION</b>The method reported in this paper can be used effectively for the quality control of total flavonoid of Fructus Aurantii.</p>
Subject(s)
Chromatography, High Pressure Liquid , Methods , Citrus , Chemistry , Flavonoids , Quality ControlABSTRACT
<p><b>OBJECTIVE</b>To investigate the correlation between the polymorphism of the tumor necrosis factor-related apoptosis inducing ligand (TRAIL) and the genetic susceptibility to prostate cancer (PCa) in the Chinese Han population in Nanjing.</p><p><b>METHODS</b>We performed a case control study on 187 cases of PCa and 237 cancer-free healthy controls. Peripheral blood genome DNA was extracted from the subjects for analysis of the polymorphism of the TRAIL-716 locus by polymerase chain reaction-ligase detection reaction (PCR-LDR). The correlations between the susceptibility to PCa and different genotypes were compared.</p><p><b>RESULTS</b>An SNP (-716A/G) was found in the promoter of the TRAIL gene. AA, AG and GG genotypes were identified. Logistic regression analysis suggested that AG, GG and AG + GG genotypes had no significant correlation with the risk of PCa (OR = 0.89, 95% CI = 0.54 -1.47; OR = 0.94, 95% CI = 0.69 -1.27; OR = 0.87, 95% CI = 0.54 - 1.41).</p><p><b>CONCLUSION</b>The TRAIL-716 polymorphism is not directly related with the genetic susceptibility to PCa in the Chinese Han population of Nanjing.</p>