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Objective:To study the regulatory effect of miR-9 on the proliferation of breast cancer cells by targeting hexokinase 2 (HK2) .Methods:Breast cancer tissues and paracancer tissues were collected and the expression levels of miR-9 and HK2 were detected. MCF-7 cells were cultured and divided into blank control group, NC group, miR-9 group, NC-siRNA group and HK2-siRNA group. The cell viability, expression levels of HK2 and cleaved caspase-3 were detected, the targeted binding of miR-9 to HK2 was verifed.Results:the expression level of miR-9 in breast cancer was lower than that in paracancer tissue (0.52±0.08 vs 1.05±0.25, t=16.685, P<0.000) , and the expression level of HK2 was higher than that in paracancer tissue (0.73±0.14 vs 0.34±0.08, t=17.587, P<0.000) , and the expression level of miR-9 was negatively correlated with HK2; the OD490 level, the expression level of HK2 and the fluorescence activity of double luciferase reporter gene containing HK2 mRNA 3 'UTR in miR-9 group were lower than those in NC group (0.58±0.09 vs 1.04±0.21, 0.51±0.08 vs 1.18±0.24, 41.11±9.28 vs 148.28±29.59, t/P=4.027/0.007, 5.297/0.002, 6.912/0.001) , and the expression level of cleaved caspase-3 was higher than that in NC group (1.08±0.26 vs 0.42±0.09, t/P=4.797/0.003) . The OD490 level and the expression level of HK in HK-siRNA group were higher than that in NC-siRNA group, and the expression level of cleaved caspase-3 was higher than that of NC-siRNA group. Conclusion:miR-9 can inhibit the proliferation of breast cancer cells, and its mechanism may be related to the targeted inhibition of HK2 expression and the increase of downstream cleaved caspase-3 expression.
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Objective To observe the effect of Rhy-SLN on the proliferation of rat vascular smooth muscle cells (VSMC) induced by TGF-β1, and explore the mechanism. Methods The primary culture of rat thoracic aortic vascular smooth muscle cells was studied by tissue block culture method. The cells were divided into the control group, TGF-β1 group, TGF-β1+ the high, medium and low dosage groups of Rhy-SLN. In addition to the control group, the cells of the other groups were involved in the intervention of TGF-β1 of 20 g/L, and the high, medium and low dosage groups of Rhy-SLN cells were involved in the intervention of 25, 50, 100 mg/L of the hook teng solid lipid nanoparticles. After 24 hours of culture, MTT assay was used to detect cell proliferation inhibition rate in each group, and the cell cycle was detected by flow cytometry. The expression of c-myc and c-Fos protein in each group was detected by Western blot method. Results Compared with the TGF-β1 group, the absorbance value (0.457 ± 0.046 vs. 0.975 ± 0.049) of TGF-β1+ rhy-sln high dose group significantly decreased (P<0.01); the number of S phase cells (15.87% ± 2.47%, 15.23% ± 1.69%, 17.02% ± 2.87% vs.38.58% ± 2.68%)of TGF-β1+rhy-sln in each dose group significantly decreased(P<0.01);The c-myc(48.65 ±3.65,50.69 ± 4.16,55.29 ± 3.67 vs.68.21 ± 3.25)and c-Fos(38.78 ± 4.25,43.56 ± 3.69,46.58 ± 3.57 vs.66.54 ± 4.09) of the TGF-β1+ rhy-sln each dose group significantly decreased (P<0.01). Conclusions The Rhy-SLN can inhibit the proliferation of VSMC in rats induced by TGF-β1.Its mechanism is related to the conversion of G0/G1 phase to the S phase and the expression of the reduction of c-myc and c-fos protein.
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Background and purpose: Multidrug resistance of tumor cells is the main factor for the failure of chemotherapy. It is found that the apigenin has the anti-tumor effect, but its role in multidrug resistant cells was rarely reported. This study aimed to investigate the effect of apigenin on multidrug resistant breast cancer cell line MCF-7/ADR, and to explore the role of apigenin in reversing multidrug resistance. Methods: The MCF-7/ADR cells were cultured with different concentrations of apigenin, and the same cells were cultured with ADR in the control group. Thecell proliferation was detected by MTT, the cell cycle distribution was detected by PI, and the cell apoptosis was detect-ed by Annexin V/PI. The drug sensitivity in vitro was detected by the method of MTT, and the drug retention rate was detected by rhodamine 123 accumulation. The expression of P-gp protein was measured by Western blot, the RT-PCR method was used to detect the transcription of multidrug resistance gene MDR1. Results: The MCF-7/ADR cell prolif-eration was inhibited by the apigenin, the cell cycle progression was blocked by the apigenin, and the cell apoptosis was induced by the apigenin. There were significant differences between the apigenin group and the ADR group (P<0.05). The IC50 of ADR on MCF-7/ADR cell was (12.37±0.18) μg/mL with the apigenin effect, while the IC50 of ADR on MCF-7/ADR cell was (39.83±0.29) μg/mL without the apigenin effect (P<0.05). The reversal index was 3.22. The retention rate of rhodamine 123 in MCF-7/ADR cells in the apigenin group was higher than that in the ADR group. The MDR1 gene transcription level in MCF-7/ADR cells was higher than that in the MCF-7 cells, and the P-gp expression in MCF-7/ADR cells was higher than that in the MCF-7 cells. However, the level of MDR1 gene transcription and P-gp expression were down-regulated by the apigenin in the MCF-7/ADR cells. Conclusion: The apigenin had anti-MCF-7/ADR effect, and played the role of reversing multidrug resistance in the MCF-7/ADR cells. The mechanism may be related to down-regulation of the MDR1 gene transcription and the P-gp mediated drug e?ux function.
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Objective To investigate the anti-inflammatory and analgesic effects of toothache drop pills. Methods The acute inflammatory models such as xylene-induced ear edema and egg white-induced paw edema and the chronic inflammatory model granuloma induced by cotton pellet implantation were used in researching the inflammatory effects of toothache drop pills. Meanwhile, the analgesic effects of toothache drop pills were observed by hot plate and acetic acid writhing test. Results Compared with the blank control group,the degree of ear swelling in mice in the positive control group, toothache drop pills middle and high dose group (2.56 ± 1.35 mg, 4.26 ± 1.21 mg, 3.23 ± 1.25 mg vs. 8.25 ± 1.21 mg) were lower than the blank control group (P<0.05). 120 minutes after administration,compared with the blank control group, the degree of paw swelling in mice in the positive control group, toothache drop pills middle and high dose group (23.54 ± 9.12 mg, 27.58 ± 9.14 mg, 21.25 ± 8.45 mg vs.39.54 ± 8.89 mg) were lower than the blank control group (P<0.05). Granuloma swelling quality in mice caused by cotton in the positive control group, toothache drop pills middle and high dose group (6.51 ± 2.58 mg, 7.82 ± 1.57 mg, 6.58 ± 3.47 mg vs. 13.58 ± 3.25 mg) were lower than the blank control group (P<0.05). The threshold of pain in mice in the positive control group, toothache drop pills middle and high dose group (20.86 ± 2.58 s, 20.25 ± 2.14 s, 20.75 ± 1.78 s vs.17.21 ± 3.31 s) were increased (P<0.05). The number of mice writhing in the positive control group, toothache drop pills middle and high dose group (23.47 ± 7.57, 28.65 ± 6.54, 24.36 ± 7.78 vs. 40.96 ± 6.58) were decreased(P<0.05). Conclusion Toothache drop pills had obvious anti-inflammatory and analgesic effects.
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Objective To prepare toothache drop pills and establish its quality standard.Methods The drop pills were prepared by a routine method; HPLC was used for the determination of the EU isoimperatorin content.Results The drop pills were well- distributed in size, smooth and glossy in appearance, mild in hardness; TLC can identify isoimperatorin characteristic spots; Determination of the Indigo 0.50~520μg/ml linear relationship was good,r=0.999 8, the recovery of 98.38%.RSD values were 1.05%.Conclusion The preparation method of toothache drop pills is simple, and the drop pills are well-shaped with controllable quality.
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ObjectiveTo explore the content of isorhynchophylline in different medicinal parts of Ramulus Uncariae Cum Uncis.MethodsHPLC was adopted to determine 8 batches of different Ramulus Uncariae Cum Uncis of different origin and different medicinal parts. The Waters Symmetry C18 color(4.6 mm×250 mm, 5μm) was used with mobile phase of methanol -0.01 mol/L ammonium acetate buffer(pH 8.0)(60∶40), column temperature of 25℃ , 20μl sample volume, velocity of 1.0 ml/min, and detection wavelength of 246 nm. ResultsIsorhynchophylline can be detected in all 8 batchs of Ramulus Uncariae Cum Uncis in different regions, and different content were found among different origins. The content of isorhynchophylline in different parts of the same origin showed a decreasing sequence of the rhabd, stem with hook, stem without hook, and twig without hook and leaves.ConclusionMost of medicinal part of Ramulus Uncariae Cum Uncis contains isorhynchophylline, which provide a lab basis for exploring medicinal parts of this herbal medicine.
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Licorice, the oldest Chinese traditional medicine, is widely used in the treatment of human diseases. Due to the deficiency of wild resource, selecting and breeding becomes a key issue to expanding the supply of licorice. Spaceflight technology will become a new method for medicinal plants. The aim of this study was to investigate the effect of spaceflight on the components and anti-inflammatory activity in licorice. After flowing on a recoverable satellite for 18 days, licorice seeds were germinated and grown to maturity and the parallel ground-based seeds were also planted under the same conditions. The main components in licorice root were analyzed through HPLC. The contents of two components in spaceflight groups were higher than that of the ground control ones. Three acute inflammatory models including xylene-induced auricular edema, carrageenan-induced paw edema and acetic acid-induced vascular permeability were utilized to compare the anti-inflammatory activity of licorice pre and post spaceflight. The licorice extract showed the significant anti-inflammation activity. After the spaceflight, the pharmacological activity of licorice got higher than that of the ground control one. All of the models gained the tendency that the spaceflight group of species Hangjinqi had the strongest activity than other groups. The research provided the scientific data for a new breeding of medicinal plant through the spaceflight and indicated that the technology of space flight may be a new effective method for the breeding and cultivation of licorice
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ObjectiveTo evaluate the significance of CK19 mRNA monitoring in peripheral blood of breast cancer patients.MethodsPeripheral blood samples were collected from 137 breast cancer patients preoperatively,one day post-operation,7 days,1,3,6,12,18,and 24 months after operation.RTPCR was used to detect CK19 mRNA expression.The relationships were analyzed between CK19 mRNA expression and treatment result, between CK19 mRNA expression and clinicopathological parameters.ResultsThere was no significant difference between the level of CK19 mRNA as tested preoperatively,and 1,7 days postoperatively(P ＞ 0.05 ).From one month and thereafter,CK 19 mRNA decreased significantly when compared with that immediate perioperatives ( P ＜ 0.05 ). Postoperative peripheral CK19 mRNA expression increased in those found with recurrent or metastasized tumors ( P ＜ 0.01 ). CK19 mRNA expression does not correlate with patient's menstrual status,estrogen receptor expression,progesterone receptor expression,HER-2 expression and Ki67 proliferation expression (P ＞ 0.05 ). But there was statistically significant correlation between CK19 mRNA expression and tumour size,histology grading,pathological type,lymph node metastasis ( P ＜ 0.01 ).ConclusionsIn breast cancer patients peripheral blood CK19 mRNA expression is correlated with risk clinicopathological parameters, and increases in patients with postoperative recurrence and tumor metastasis.
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Objective To study the expression of high molecular weight cytokeratin (CK34βE12) in breast carcinoma and the relationship between the high molecular weight cytokeratin expression and patient's prognosis. Methods We detected the high molecular weight cytokeratin expression in 100 patient's of breast cancer tissues and paracancerous breast tissue in 30 cases immunohistochemically. The relationship between the expression of high molecular weight cytokeratin in breast cancer tissues and age, menstrual status, estrogen receptor expression, progesterone receptor expression, histology grading, TNM staging, pathology type, and patient survival was analyzed. Results (1) There was no significant correlation between high molecular weight cytokeratin expression of breast -cancer tissues and age, menstrual status, estrogen receptor expression, progesterone receptor expression (all P>0.05). But the correlation was statistically significant between that expression and histology grading, TNM staging, pathology category (all P<0.05). (2) By univariate analysis, the 5-year disease-free survival rate in patients with negative expression was lower than those with positive expression(P<0.05). By multivariate analysis, the expression of high molecular weight cytokeratin was unfavorably related with tumor free survival. Conclusion The expression of high molecular weight cytokeratin in breast carcinoma tissues was correlated with biologic malignancies of breast cancer, the negative high molecular weight cytokeratin expression of breast cancer tissues predicts poor prognosis.
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Objective To explore the expression of COX-2,PD-ECGF and VEGF in gastric carcinoma tissue,and analyze and identify the relationship between the expression and tumor angiogenesis together with biological behaviour in gastric carcinoma.Methods ElivisionTM immunohistochemical method was performed to detect the expression of COX-2,PD-ECGF,VEGF and CD34,and evaluate the value of MVD in surgically resected gastric carcinoma specimens and biopsy tissues under gastroscope,including normal gastric mucosa tissues, metaplasia tissues and gastric mucosal atypical hyperplasia tissues. Clinicopathologic data of patients were analyzed retrospectively. Results The positive expression of COX-2, PD-ECGF,VEGF and MVD in gastric carcinoma were significantly higher than those in normal gastric mucosa, metaplasia or gastric mucosal atypical hyperplasia,and also their expression and MVD were closely related to lymph node metastasis and depth of invasion. The MVD in the groups of COX-2,PD-ECGF,VEGF positive-expression were significantly higher than those in negative-expression groups respectively. There were positive correlations among the expression of COX-2,PD-ECGF and VEGF in gastric carcinoma. MVD increased significantly when COX-2, PD-ECGF and VEGF coexpressed. Conclusion The expression of COX-2,PD-ECGF,VEGF and tumor angiogenesis participated in the genesis,invasion and metastasis of human gastric carcinoma.COX-2,PD-ECGF and VEGF participated in the tumor angiogenesis of gastric carcinoma. In the process of tumor angiogenesis and expression in gastric carcinoma, COX-2, PD-ECGF and VEGF could strengthen the effectiveness each other.They could strengthen the effective ness of tumor angiogenesis when they coexpressed. They could be selected as targets for tumor anti-angiogenesis treatment.
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Objective To investigate the localization and distribution of the five endocrine cells in the digestive tract mucosa of ricefield eel(Monopterus albus). Methods Using immunocytochemical technique of strept avidin-biotin-peroxidase complex(SABC) were used. Results At least 5 kinds of immunoreactive endocrine cells distributed in the digestive tract mucosa of M.albus. They were gastrin(Gas),somatostatin(Som),5-hydroxytryptamine(5-HT),insulin(Ins),and neurofilament (NF) immunoreactive endocrine(IRE) cells.Gas and Som-IRE cells distributed between stratified squamous epithelium and goblet cell in esophagus. A large number of Gas-IRE cells were found between gastric fundus epithelium and gastric glands, and only a few in the carcia. Ins, 5-HT and NF-IRE cells distributed in the epithelium pylorus and pyloric glandular tube respectively. No any immunoreactive positive reaction was found in the gut of M.albus.In addition, immunoreactive positive reaction of glucagons was not found in whole digestive tract.All immunoreactive endocrine cells were dark brown in color.Their morphology was irregular, cytoplasmic process was shorter and thicker, their nucleus showed an empty bubble.They distributed between esophageal epithelium and gastric epithelium or glandular epithelium, and cytoplasmic process extended to the gastric lumen and glandular cavity.Conclusion There is a complex endocrine function of the digestive system in ricefield eel (M.albus) at the lowest vertebrate.