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【Objective】 To investigate the effect of N-acetylcysteine （NAC） on mitochondrial damage of airway epithelial cells induced by cigarette smoke extract （CSE）. 【Methods】 Human airway epithelial cells （BEAS-2B） were cultured and divided into three groups as follows： normal control group， 7.5% （75 mL/L） CSE-treated group and 7.5% CSE plus NAC group. After stimulation for 24 hours， cell viability was determined by MTT， and the levels of mitochondrial reactive oxygen species （ROS） and mitochondrial membrane potential （MMP） were observed under the fluorescence microscope. MMP was also measured by flow cytometry， the protein expressions of Sirt3 and manganese superoxide dismutase （MnSOD） were detected by Western blotting， and MnSOD activity was measured by colorimetry. 【Results】 Pretreatment with NAC significantly improved the viability of airway epithelial cells （P<0.05）. The results of fluorescence microscopy and flow cytometry showed that NAC pretreatment significantly attenuated MMP decline in airway epithelial cells exposed to 7.5% CSE （P<0.05）. Compared with 7.5% CSE-treated group， mitochondrial ROS in airway epithelial cells was significantly decreased in 7.5% CSE plus NAC group （P<0.05）. In addition， pretreatment with NAC significantly inhibited the decrease of Sirt3 and MnSOD protein expression and improved MnSOD activity in airway epithelial cells exposed to 7.5% CSE （P<0.05）. 【Conclusion】 NAC attenuates CSE-induced airway epithelial mitochondrial damage through the regulation of Sirt3-MnSOD signaling pathway， which reveals a new mechanism of NAC treatment for chronic obstructive pulmonary disease.
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BACKGROUND: It has been widely reported that the most types of cancer are probably originated from cancer stem cells (CSCs) which are subpopulations of tumor cells. Recent studies also suggested that lung cancer could arise from CSCs. However, the phenotypic characteristics of CSCs in lung cancer have not been precisely described.OBJECTIVE: To systematically analyze the expression of the most common CSC markers in cell line A549.METHODS: A549 cells were cultured for 1 week under the condition of conventional high glucose. After that, flow cytometry was used to assess the expression of putative stem cell markers, including CD133, CD24, CD44 and ABCG2.Cells were then sorted according to the expression of CD44 and CD24 markers by fluorescence-activated cell sorting (FACS) and characterized using their sphere-forming capacity in serum free medium supplemented with several growth factors.RESULTS AND CONCLUSION: A549 cells expressed the CSC markers CD44 and CD24 at 64.23％ and 58.62％,whereas the expression of both ABCG2 and CD133 was around 0.9％. Double-positive CD44/133 populations were rare.CD44+/CD24+ and CD44+/CD24- subpopulations respectively exhibited 54.64％ and 23.38％ expression. CD44+/CD24+ and CD44+/CD24- subsets were sorted by FACS. Both isolated subpopulations formed spheres in the serum free medium supplemented with basic fibroblast growth factor and epidermal growth factor. However, there was no significant difference in the sphere formation efficiency among CD44+/CD24+ and CD44+/CD24- subsets as well as A549 cells. Our findings suggest that CD44 and CD24 cannot be considered as potential markers for isolating lung CSCs in cell line A549, and further investigation using in vivo assays is required.
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Objective To investigate the role of Wnt/β-catenin pathway in regulating allergic airway inflammation in asthmatic mice.Methods We induced dendritic cells (DCs) from bone marrow of BALB/c mice,and then treated the cells with LiCl and PKF118-310,separately.We observed the morphological features of DCs under light microscope.Mixed lymphocyte reaction (MLR) was used to observe the functional changes of DCs.Western blot was used to detect the expressions of GSK-3β and β-catenin at the protein level.We established a mouse asthma model by using ovalbumin (OVA),and then treated these mice with LiCl and PKF118-310.The total number of cells and eosinophil percentage in BALF were determined.The lungs of mice were observed by HE staining to evaluate the degree of allergic inflammation.The cytokines in BALF and spleen cells supernatant were assayed by enzyme-linked immunoassay (ELISA),and the total IgE in the serum was also measured by ELISA.The protein expression levels of GSK-3β and β-catenin in lung tissue were assayed by Western blot.Results ① The DCs treated with LiCl promoted the proliferation of allogeneic T lymphocytes in MLR more weakly than those treated with PKF118-310 (P＜0.01).② The GSK-3β protein expression level of DCs treated with LiCl was significantly lower than DCs treated with PKF118-310.In contrast,the β-catenin protein expression of DCs treated with LiCl was higher than that of DCs treated with PKF118-310 (P ＜ 0.01).③ The total number of cells and eosinophil percentage in BALF were significantly increased in the experimental group compared with those in the control group (P＜0.01).There was also a significant difference between LiCl group and PKF118-310 group (P＜0.01).④ In the three experimental groups,the severity of inflammation in the lungs of LiCl group was weaker than that in PKF118-310 group (P＜0.05).⑤ Compared with that in the normal control group,IL-4 in BALF and spleen cell culture supernatant of the experimental group was significantly higher while IFN-γ was the opposite (P＜0.01).LiCl group had the lowest level of IL-4 and the highest level of IFN-γ;PKF groups was the opposite (P＜0.05).⑥ The total IgE in serum was significantly increased in the experimental group compared with the control group (P＜0.01).There was also a significant difference between LiCl group and PKF118-310 group (P＜0.05).⑦ GSK-3β protein expression was significantly lower in LiCl group than in PKF118-310 group (P＜0.05),while β-catenin protein expression was significantly higher in LiCl group than in PKF118-310 group (P＜0.05).Conclusion LiCl and PKF118-310 can affect the severity of asthma by regulating Wnt/β-catenin signal pathway and the expressions of GSK-3β andβ-catenin protein,which provides a new direction for asthma treatment.
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Objective To summarize the clinical nursing experience for patients with malignant obstructive jaundice after receiving percutaneous biliary radiofrequency ablation (RFA) and stent implantation. Methods The postoperative nursing experience in 9 patients with malignant obstructive jaundice who received percutaneous biliary RFA together with stent implantation were retrospectively analyzed. The postoperative complications and the nursing intervention measures were analyzed and evaluated. Results Biliary RFA and subsequent stent implantation were successfully carried out in all 9 patients. After the procedure, biliary fistula occurred in one patient, biliary hemorrhage in 2 patients and biliary infection in 2 patients. The patient, who developed biliary fistula, died one week later, and the clinical conditions in the remaining 4 patients were improved after symptomatic treatment. During the follow-up period of one month, the patients were in good condition. Conclusion The main purpose of postoperative nursing for patients after receiving percutaneous biliary RFA is to prevent the occurrence of bile duct perforation, hemorrhage, infection, etc. It is very important to keep the patients under close observation and comprehensive nursing so as to make an early detection and timely treatment of such complications, thus to reduce the incidence of complications causing serious consequences as well as to promote an early recovery.
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Objective To develop a rotatable scalp acupuncture to enhance success rate of arteriovenous puncture during hospital treatment, field firstaid and evacuation.Methods The acupuncture was composed of a pinhead, a neilsbed, wings and a soft tube. The wings connected with each other to form a hole to hold the intermediate section of the neilsbed rotating in it. There was an index point in the middle of the head of the neilsbed, whose direction corresponded linearly with that of the inclined plane of the pinhead in the vessel.Results Clinical trials proved that the acupuncture could eliminate the failure of puncture due to tremble hands, incoordination of the patient and etc.Conclusion The acupuncture structured well is easy to operate for all conditions, and thus is worth popularizing practically.
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Nitrogen source is one of the important factors that affect the microalgae growth and lipid accumulation. We studied the effects of various nitrogen sources (i.e. NaNO3, CO(NH2)2, NH4Cl and CH3COONH4) and amount on the growth density, lipid yield, and eicosapentaenoic acid (EPA) content of Nannochloropsis oculata by single factor experimental method. The results show that N. oculata preferred NH4+ as nitrogen source rather than NO3- and CO(NH2)2. NH4+ could promote the growth and lipid accumulation of N. oculata. With the increase of nitrogen concentration, the biomass and the content of polyunsaturated fatty acid (PUFA) increased, but the content of lipid decreased. CH3COONH4 was the most suitable for growth, accumulation of lipid and EPA of N. oculata among the four investigated nitrogen sources. The optimal concentration was 5.29 mmol/L.
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Eicosapentaenoic Acid , Metabolism , Lipids , Microalgae , Metabolism , Nitrogen , Metabolism , Stramenopiles , MetabolismABSTRACT
Objective A HPLC method to determine the content of Baicalein in Tangtai capsule was established. Method The separation was performed by Zorbax C18 column with acetonitrile and 0.5% phosphorice acid. The elution program was 1~15 min, 15%～60% acetonitrile. The flow rate was 1 mL/min and column temperature was 25 ℃. Content was detected at a wavelength of 280 nm. Result The linear range was 0.936～18.72 ?g, regression:Y =4849.53X-0.9593 (r=0.9999). The average recovery rate was 98.29% and RSD was 0.97% (n =5). Conclusion The method is simple, accurate and reproductive, and can be used for determining Baicalein in Tangtai capsule.
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Objective: To establish the quality criteria for xiaozhining tablet. Methods: TLC method was applicated to determine and differentiate the Scutellaria , the Radix salviae miltiorrhizae, the Rheum, the Leonurus of the tablet. HPLC method was used for detecting the content of emodin in xiaozhining Tablet. Results: Four main components of the tablet was determined well. The regression equation by HPLC was A=2.43?10 7C+6.06?10 4(r=0.999 7). Curcumin had good linear relation within the range of 0.5～0.05 mg/ml. The average recovery rate was 96.70%(RSD=1.06%). The average value of emodin was 0.021 4 mg. Conclusion: The method of determination is reliable. It can be used as the basis for the quality control of xiaozhining Tablet.
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AIM: To develop a method of determining notoginsenoside R_1,ginsenoside Rg_1 and Rb_1 contents in Xinnaozhi Capsule(Radix et Rhizoma Notogineseng,Hirudo,Eupolyphaga seu steleophage). METHODS: HPLC was used to determine notoginsenoside R_1,ginsenoside Rg_1 and Rb_1 in Xinnaozhi Capsule.The separation was performed by Zorbax C_(18) colunm with acetonitrile and water using a gradient program at UV 203nm at room temperature.The elution program was 0-3 min,20%-25% acetonitrile,3-15 min,25%-45% acetonitrile. RESULTS: 3 saponins were separated well.Average recoveries were 102.93% for notoginsenoside R_1(RSD=(1.26%));105.68% for ginsenoside Rg_1(RSD=1.52%);104.34% for ginsenoside Rb_1(RSD=0.70%),respectively. CONCLUSION: The method is simple and rapid and with satisfactory results.It is suitable for quality control of Xinnaozhi Capsules.
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AIM: To establish a quantitative analytical method of ferulic acid in Kexintong Dripping Pills (Moschus, Rhizoma Chuaxiong, Rhizoma Corydalis, etc.) based on the capillary zone electrophoresis. METHODS: P-Nitrobenzoic acid was adopted as the internal substauce with a 30 mmol?L -1 borax solution at equal electrophoresis current of 50 ?A, detection wavelength at 295 nm, uncoated fused silica capillary of 75 ?m?50 cm, temperature at 20 ℃. Sample was injected for 5 s at pressure of 20PSI. RESULTS: Good linearity was showed in the concentration range of 4～264 mg?L -1 , RSD was 1.71%(n=6), and the recovery was 100.47%?2.1%. CONCLUSION: The result indicates the proposed method is convenient, rapid, accurate and reliable, it is a proper method for the quantitative analysis of ferulic acid in Kexintong Dripping Pills.