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Objective:To investigate whether chlorogenic acid can inhibit the proliferation, migration, invasion and promote apoptosis of lung cancer A549 cells by causing mitochondrial dysfunction through PI3K-Akt pathway.Methods:A549 cells were treated with chlorogenic acid at concentrations of 0, 25, 50, 100, 150, and 200 μg/ml for 48 h. CCK-8 assay was used to detect the cell proliferation rate and calculate the half maximal inhibitory concentration (IC 50). A549 cells were divided into three groups: control group, chlorogenic acid group (IC 50) and chlorogenic acid + 740-YP group (IC 50 chlorogenic acid +50 μg/ml 740YP). After 48 h of intervention, the cell migration distance was detected by cell scratch assay. Cell invasion assay was used to detect cell invasion ability. Cell cycle, apoptosis and mitochondrial membrane potential were detected by flow cytometry. The content of malondialdehyde (MDA) in cell supernatant was detected by enzyme-linked immunosorbent assay (ELISA). Western blotting was used to detect the protein expression of p-PI3K, p-Akt and Caspase3. Results:The IC 50 of chlorogenic acid to A549 cells was 57.45 μg/ml. The results of cell scratch assay showed that the 48 h migration distances of the control group, chlorogenic acid group and chlorogenic acid + 740YP group were (424.80±14.43), (289.67±18.93) and (402.22±17.99) μm, respectively. The results of cell invasion assay showed that the numbers of invasive cells after 48 h were 96.00±6.24, 35.33±7.64 and 83.00±2.00, and the results of flow cytometry showed that the 48 h apoptosis rates were (6.15±0.17) %, (54.63±0.72) % and (17.27±0.39) %, respectively, among the three groups with statistically significant differences ( F=105.98, P<0.001; F=90.62, P<0.001; F=8 321.99, P<0.001). Compared with the control group, the cell migration distances and invasive numbers of chlorogenic acid group and chlorogenic acid + 740YP group were decreased (all P<0.05), while the apoptosis rates were significantly increased (both P<0.001). Compared with chlorogenic acid group, the cell migration distance of chlorogenic acid + 740YP group increased ( P<0.001), the number of cell invasion increased ( P<0.001), and the apoptosis rate decreased ( P<0.001). The results of flow cytometry showed that the proportions of cells in G 0/G 1 phase in the control group, chlorogenic acid group and chlorogenic acid + 740YP group were (65.75±0.58) %, (55.84±0.78) % and (55.24±1.37) %, respectively. The proportions of G 2/M phase were (11.21±1.03) %, (20.23±0.62) % and (9.96±0.33) %, and the proportions of S phase were (23.04±0.49) %, (23.92±1.36) % and (34.80±1.15) %, respectively, with statistically significant differences ( F=111.02, P<0.001; F=181.26, P<0.001; F=113.05, P<0.001). Compared with the control group, the proportions of G 0/G 1 phase cells in chlorogenic acid group and chlorogenic acid + 740YP group decreased (both P<0.001), and the proportion of G 2/M phase in chlorogenic acid group increased ( P<0.001), and the proportion of S phase cells in chlorogenic acid + 740YP group increased ( P<0.001). Compared with chlorogenic acid group, the proportion of G 2/M phase cells decreased and the proportion of S phase cells increased in chlorogenic acid + 740YP group (both P<0.001). The results of mitochondrial membrane potential detection showed that the JC-1 fluorescence intensity of mitochondria in the control group, chlorogenic acid group and chlorogenic acid + 740YP group were 39.51±1.32, 10.05±0.19 and 21.85±1.45, respectively, with a statistically significant difference ( F=508.82, P<0.001). Compared with the control group, the fluorescence intensity of chlorogenic acid group and chlorogenic acid + 740YP group decreased (both P<0.001). Compared with chlorogenic acid group, the fluorescence intensity of chlorogenic acid + 740YP group increased ( P<0.001). ELISA results showed that the MDA contents of the control group, chlorogenic acid group and chlorogenic acid + 740YP group were (0.47±0.01), (0.61±0.01) and (0.56±0.01) nmol/ml, respectively, with a statistically significant difference ( F=162.30, P<0.001). Compared with the control group, MDA contents in chlorogenic acid group and chlorogenic acid + 740YP group increased (both P<0.001). Compared with chlorogenic acid group, MDA content in chlorogenic acid + 740YP group decreased ( P=0.001). Western blotting results showed that the relative protein expression levels of p-PI3K in the control group, chlorogenic acid group and chlorogenic acid + 740YP group were 1.01±0.33, 0.28±0.14 and 0.34±0.20, respectively. The relative protein expression levels of p-Akt were 1.00±0.16, 0.43±0.05 and 0.95±0.14, and the relative protein expression levels of Caspase3 were 1.00±0.04, 1.41±0.05 and 0.70±0.13, respectively, and there were statistically significant differences ( F=8.48, P=0.018; F=19.11, P=0.002; F=57.50, P<0.001). Compared with the control group, the expressions of p-PI3K and p-Akt protein in chlorogenic acid group decreased, and the expression of Caspase3 protein increased (all P<0.05). The expressions of p-PI3K and Caspase3 protein in chlorogenic acid + 740YP group decreased (both P<0.05). Compared with chlorogenic acid group, the expression of p-Akt protein in chlorogenic acid + 740YP group increased, and the expression of Caspase3 protein decreased (both P<0.05) . Conclusion:Chlorogenic acid may inhibit the PI3K-Akt pathway by reducing the phosphorylation of PI3K and Akt proteins, resulting in the damage of mitochondrial function and the accumulation of MDA, which eventually leads to the damage of lung cancer A549 cells function and the reduction of cells activity, and then promotes cells apoptosis.
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Objective@#To evaluate the application of FISH testing of bcl-2/IgH gene translocation and IgH/L gene rearrangement in different stages of follicular lymphoma.@*Methods@#In 32 follicular lymphoma cases, which were collected at Guangdong General Hospital from September 2014 to December 2016, the bcl-2/IgH gene ectopic state was detected by FISH while the IgH/L gene rearrangement was tested using PCR-GeneScan to analyze the relationship between bcl-2/IgH gene translocation, different stages of follicular lymphoma and clonal immunoglobulin (IgH/L) gene rearrangements.@*Results@#From the paraffin sections of all 32 follicular lymphomas, 17 cases showed bcl-2/IgH gene translocation, and the percentages of FL1, FL2 and FL3 translocation were 12/13, 3/5 and 2/14, respectively. Among the 24 cases of IgH/L gene arrangements identified from the total sample, the occurrence rates of FL1, FL2 and FL3 gene arrangement were 7/13, 4/5 and 13/14, respectively. Spearman′s rank correlation analysis and χ2 analysis showed that bcl-2/IgH gene translocation was negatively correlated with follicular lymphoma stage and the association was statistically significant. In more advanced stages of follicular lymphoma, the occurrence of bcl-2/IgH gene translocation tended to decrease with distinct FL1, FL2 and Fl3 gene expression (P<0.05). As IgH/L gene rearrangement in FL3 was higher than that in FL1 and FL2, its detection may be complimentary to FISH test for bcl-2/IgH gene translocation in diagnosing follicular lymphoma.@*Conclusions@#The combined use of FISH and PCR-GeneScan increases the positive rate of follicular lymphoma diagnosis, and this combination is more sensitive than FISH or clonal analysis only to detect the chromosomal abnormality or the gene rearrangement.
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<p><b>OBJECTIVE</b>To investigate mutations frequencies of KRAS,NRAS and BRAF genes in colorectal carcinoma.</p><p><b>METHODS</b>Tissue specimens from 200 colorectal cancer patients at diagnosis were collected and subject to KRAS,NRAS and BRAF mutation analyses by PCR-based direct DNA sequencing targeting exons 2, 3 and 4 of KRAS gene, exons 2, 3 and 4 of NRAS gene and exon 15 of BRAF gene.</p><p><b>RESULTS</b>Activating mutations were detected in KRAS (44%, 88/200), NRAS (2%, 4/200) and BRAF (5%, 10/200) in this study cohort.Among KRAS mutations, 64.8% (57/88) occurred in codon 12 and 12.5% (11/88) occurred in codon 13. KRAS gene mutation in exon 3 mainly involved codons 59 and 61. KRAS gene mutation in exon 4 mainly involved codons 117 and 146.</p><p><b>CONCLUSIONS</b>Mutations at exon 2 of KRAS gene have the highest frequency in colorectal carcinoma. Expanding the detection sites of KRAS gene combined with NRAS and BRAF genes may help to identify patients who will most likely benefit from targeted therapies.</p>
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Feminino , Humanos , Sequência de Bases , Códon , Neoplasias Colorretais , Genética , Análise Mutacional de DNA , Éxons , Genes ras , Mutação , Proteínas Proto-Oncogênicas , Proteínas Proto-Oncogênicas B-raf , Genética , Análise de Sequência de DNARESUMO
As the dressings currently used in clinic settings unflat shape in general, they can't be fitted completely on occiput, heel, elbow, knee and other body parts unflat. This paper developed one kind of foam dressing of special shape based on local anatomy. The foam dressing is waterproof and air permeable, it can cover the wound closely enough to prevent bacteria from invasion and infection. With a saturated absorption ratio of 1: 8 or higher, it can keep the wound clean and moisture by absorbing large amounts of wound inflammatory secretions and is almost completely permeable to oxygen and carbon dioxide. Assuring safety and effect meanwhile, it has better outcomes than common dressings in the same application settings.
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Bandagens , CicatrizaçãoRESUMO
<p><b>OBJECTIVE</b>To compare the value of applicability of the 7th edition of UICC-AJCC esophageal and gastric cancer TNM staging system in the prognostic prediction of adenocarcinoma of esophagogastric junction (EGJ).</p><p><b>METHODS</b>During June 1, 2007 through Dec. 31, 2010, a total of 199 patients with adenocarcinoma of esophagogastric junction (Siewert type II) underwent R0-intent resection from June 1, 2007 to Dec 31, 2010 in our hospital. Their clinicopathological and survival data were retrospectively analyzed with Kaplan-Meier and Cox regression models. They were restaged according to the 7th edition of UICC/AJCC TNM stage systems for esophageal adenocarcinoma and gastric cancer, respectively. Then the likelihood ratio chi-square test related to the Cox regression model and Akaike information criterion (AIC) were used for measuring goodness of fit for both staging systems.</p><p><b>RESULTS</b>199 patients with Siewert type II esophagogastric junction adenocarcinoma were identified in this study. Out of them, there were 162 males and 37 females. Their age range was from 38 to 79 years, with a median age of 62 years. 176 cases underwent transthoracic surgery, and other 23 cases underwent transabdominal surgery. TNM-EC and TNM-GC classified 4 patients to stage T1, 39 to T2, 139 to T3, and 17 to T4a, respectively, and classified 76 patients to stage N0, 58 to N1, 49 to N2, 16 to N3, respectively. The median follow-up period was 30 months. The 1-, 3-, and 5-year survival rates were 95.0%, 52.7% and 39.2%, respectively. Univariate analysis indicated that age at surgery (P = 0.009), surgical approach (P = 0.002), cell differentiation (P = 0.030), preoperative co-morbidity implications (P = 0.026), depth of tumor invasion (P < 0.001) and number of metastatic lymph nodes (P < 0.001) were significantly influencing factors of postoperative overall survival. Multivariate analysis showed that the independent prognostic factors for adenocarcinoma of esophagogastric junction were only T stage, N stage and preoperative co-morbidity and morbidities according to the 7th edition of esophageal cancer or gastric cancer TNM staging systems. The AIC value was 961.4 for the 7th edition of esophageal adenocarcinoma caner staging system, and 965.7 for the 7th edition of gastric cancer staging system.</p><p><b>CONCLUSIONS</b>The UICC/AJCC 7th edition of esophageal adenocarcinoma cancer TNM classification staging system is superior to the 7th edition of gastric cancer TNM staging system for adenocarcinoma of esophagogastric junction.</p>
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Adulto , Idoso , Idoso de 80 Anos ou mais , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Adenocarcinoma , Diagnóstico , Neoplasias Esofágicas , Diagnóstico , Junção Esofagogástrica , Linfonodos , Metástase Linfática , Estadiamento de Neoplasias , Prognóstico , Modelos de Riscos Proporcionais , Estudos Retrospectivos , Neoplasias Gástricas , Taxa de SobrevidaRESUMO
Object To observe the effect of Guizhi Fuling Capsule (GFC) on prostatic hyperplasia in rats induced by testosterone propionate. Methods Seven days after the male rats were castrated, testosterone propionate (3 mg/kg) was sc injected in rats. Meanwhile, the rats were administered GFC (2.16, 1.08, 0.54 g/kg) once a day, and the treatment continued for 30 d. An hour after the final administration, the rats were put to death, and the weight and volume of prostates were measured. The construction of prostate cells was also observed under light microscope. Results In the GFC group, the weight and volume of prostate was significantly decreased compared with that in the model group (P
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Objective To study the mechanism of blood-lipid regulation of the effective fractions from the extracts of Radix Salviae Miltiorrhizae(RSM)or Fructus Crataegi(FC),as well as their interaction.Methods The serum-drug of RSM and FC extracts were prepared by orthogonal design L_16(215).Amphotercin B cell model and RT-PCR method were used to examine the effects of the serum-drug of RSM and FC extracts on endogenous cholesterol biosynthesis in Chinese hamster ovary(CHO)cells and the expressions of cholesterol 7?-hydroxylase(CYP7A)mRNA in BRL cells.Results Ethanol(75%)and ammonia extracts of RSM obviously increased the absorbance(A)in the experiment of amphotercin B cell model and the expression of CYP7A mRNA in BRL cell line.The extracts of FC had no direct effect on cholesterol metabolism by themselves,but they were in synergism with the extracts of RSM.ConclusionThe extracts of RSM have a blood-lipid regulation by inhibiting endogenous cholesterol biosynthesis and increasing CYP7A mRNA expression,and maybe have a better effect when the extracts of FC are used together.