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Objective: To optimize the supercritical CO2 extraction process of active ingredients from Magnoliae Officinalis Cortex (MOC) and explore the antioxidant activity of the extracts. Methods: The content of magnolol and honokiol of the supercritical CO2 extracts of MOC was determined by HPLC, and the extraction process was optimized by orthogonal experiment. The antioxidant activities of the extracts were determined by MTT. Results: The optimum extraction pressure of magnolol was 25 MPa, the extraction temperature was 55 ℃, the amount of CO2 was 30 kg, and the optimum extraction parameters mentioned above of honokiol were 15 MPa, 50 ℃, and 25 kg, respectively. Conclusion: Under the optimum extraction conditions, magnolol and honokiol have high extraction efficiency, good repeatability, stability and feasibility, and the extract have good antioxidant activity.
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Objective@#To provide reference for clinical screening of individualized therapy by detecting the drug susceptibility of primary tumor cells derived from malignant pleural effusion (MPE) of the patients with non-small cell lung cancer (NSCLC). @*Methods@#MPE specimens were collected from 20 patients diagnosed as NSCLC by histopathology. They were separated by density gradient centrifugation cultured in vitro to remove non-tumor cells, and then primary cell lines were established. The half-inhibitory concentrations (IC 50 ) of conventional anti-tumor chemotherapy drugs on primary tumor cells were determined by the MTT method, and an absolute predictive value (R) was obtained by comparing the IC 50 with the theoretically calculated value of maximum plasma concentration (IC 50m ). Last, the R value was compared with the actual clinical efficacy of the NSCLC patients. @*Results@#After 20 MPE samples were pretreated and cultured for 4 generations, the primary tumor cell lines passaged stably in vitro were established successfully. Papanicolaou staining confirmed that these tumor cell lines had the characteristics of cancer cells, and their purity was nearly 100% under the microscope. The MTT results showed that the invalid IC 50 values beyond the upper limit of test concentration could be further included in the following evaluation on the therapeutic effect of patients when R values were used. When R values between 0.5 and 2.0 and more than 2.0 were used for predicting the actual therapeutic effect as disease stability and disease progression, respectively, the overall consistency between R value and actual therapeutic effect was 77% (10/13) in six patients with complete history of chemotherapy. @*Conclusion@#The primary culture of tumor cells in MPF and the detection of drug sensitivity have the clinical application value in predicting the actual therapeutic effect of NSCLC patients.
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Objective: In order to further study the chemical constituents of Blumea balsamifera and evaluate the cytotoxic activities of the compounds. Methods: The chemical components of B. balsamifera were purified by various chromatographic methods. MTT method was employed to screen the cytotoxic activities to the five tumor cells (HeLa, MCF-7, A549, MGC-803 and COLO-205 cells) of the purified compounds. Results: Two sesquiterpenoid esters blumeaene N (compound 1) and blumeaene F (compound 2) were isolated from B. balsamifera, of which, blumeaene N was a novel compound. The screening results of cytotoxin activities showed that compound 1 had certain inhibitory activities for the five tumor cells with the IC50 value ranges from 48.730 μmol/L to 97.907 μmol/L; Compound 2 also had a weak inhibitory activity on MCF-7 cell lines with the IC50 value of 91.188 μmol/L. Conclusion: Compound 1 is a new sesquiterpenoid ester found from B. balsamifera. Cytotoxic activities screening results indicated that B. balsamifera contains anti-tumor active ingredients.
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Objective:To investigate the effect of the leaching liquids of the pure titanium porcelain crowns with barium titanate coating on the proliferation of L929 cells,and to evaluate its cytotoxicity level.Methods:The L929 ceils were cultured in vitro with leaching liquids of titanium porcelain specimens with barium titanate coating (group A),and titanium porcelain specimens(group B) for 1,3 and 5 days respectively.RPMI1640 containing 10％ fetal bovine serum and 1％ phenol was served as the negative(group C) and positive control group(group D),respectively.MTT method was used to test the effects of barium titanate coating on the proliferation of mouse fibroblast L929 cells,the cytotoxicity of the 4 groups was graded.Results:L929 cells of the group showed normal morphology and vigorous growth except group D.During the whole experiment,the absorbance values of group A was greater than that of group C(P ＜0.05).The cytotoxic gradation of group A grade 0.Conclusion:Titanium porcelain specimens with barium titanate coating has a good cytocompatibility.
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Objective To study the appearance of skin mucous glycoprotein in vitro on the activity of human lung cancer cells A549. Methods Used alkali extraction and DEAE-52 ion exchange chromatography and Sephadex G-100 gel chromatography purification salamander mucous glycoprotein; Salamander mucous glycoprotein inhibition of human lung cancer cells A549 was detected by MTT colorimetric method in vitro.ResuIts It showed that the total sugar content in the appearance of mucus was 4.23%, the relatively pure glycoprotein component, by SDS protein electrophoresis tests, it contained a single glycoprotein component, its molecular weight was about 30 kDa.With glycoprotein pure concentration increased from 1,10, 20,40μg/mL, the glycoprotein inhibition rate of A549 cells increased; when the glycoprotein concentration was 40 μg/mL, for 24 h action, the inhibition rate of A549 cells was up to 85.66 %, while the role of 48 h, the inhibition rate of A549 cells was up to 92.32%.Inhibition effect of mucus glycoprotein on A549 cell compared with positive control drug, had significant difference.ConcIusion Salamander mucus glycoproteins of human lung cancer cells has obvious inhibitory effect, can provide theoretical basis for the development of lung cancer drug resistance.
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Objective: To prepare baicalin-loaded TPGS nanomicells (BCN-TPGS-PMs) and to evaluate its physicochemical properties, in vitro release behavior, and antitumor activity against MCF-7 cells. Methods: BCN-TPGS-PMs were prepared by film-thin hydration method. The preparation methods and formulations were optimized and screened based on particle size and encapsulation efficiency (EE) of micelles. The transmission electron microscope (TEM) was used to observe the particle appearance, zetasizer instrument was used to detect the diameter and Zeta potential, and ultracentrifugation was utilized to determine the EE and drug-loading rate. Dynamic dialysis method was used to study the in vitro release behavior of BCN-TPGS-PMs, and the antitumor activity against MCF-7 cells was determined by MTT method. Results: The optimal BCN-TPGS-PMs were round with the nanometric size of (11.91 ± 0.14) nm, high EE rate of (95.83 ± 7.34)%, and drug-loading rate of (5.42 ± 0.04)%. The in vitro release behavior showed that BCN-TPGS-PMs had a slow release. Compared with free BCN, BCN-TPGS-PMs showed stronger cytotoxicity and inhibition against MCF-7 cells (P < 0.05). Conclusion: The prepared BCN-TPGS-PMs have small particle size, high drug-loading rate, and good stability, and could obviously increase the in vitro inhibitory effect of BCN.
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Objective: To prepare cucurbitacin B phospholipids complex (CuB-PLC) and evaluate its physicochemical properties and in vitro antitumor activity. Methods: CuB-PLC was prepared using solvent evaporation method and optimized by Box-Behnken design. The oil-water partition coefficient, particle size, and morphology of CuB-PLC were investigated; X-ray diffraction (XRD) spectroscopy and infrared (IR) spectroscopy were used to analyze the formation machenism of CuB-PLC. MTT method was used to determine the in vitro antitumor activity of CuB-PLC. Results: The optimal formulation protocol for CuB-PLC was as follows: Tetrahydrofuran was taken as the reaction medium, phospholipids-cucurbitacin B molar ratio, reaction concentration of cucurbitacin B, reaction temperature and time were 1:1, 1.5 mg/mL, 60℃, and 3 h, respectively. The complex rate and particle size for the optimized CuB-PLC was 97.15% and (521.30 ± 10.50) nm, and the polydispersity index (PDI) was 0.133 2 ± 0.024 0. MTT experiments showed that the half of the HepG-2 cell proliferation inhibition concentration (IC50) values of CuB and CuB-PLC were 42.55 and 27.61 μmol/L. Conclusion: CuB-PLC is successfully developed under the optimized protocol, possessing high complex rate, and enhanced solubility in water, and the inhibition on HepG-2 cell proliferation is significantly enhanced, which provides the reference for the further research of CuB.
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Objective:To evaluate the cytotoxicity of colorized zirconia ceramics.Methods:Zirconia ceramic 3Y-TZP blocks were colorized by NH4 VO3 ,FeCl3 ,Co(NO3 )3 ,Ni(NO3 )3 and MoCl3 respectively.The cytotoxicity of colorized zirconia to L-929 cells was examined by immersion method and MTT assay.Results:During 3 days culture,L929 cells in all groups showed similar prolifera-tion and normal morphology.The toxicity gradation of all groups was 0 -I.Conclusion:Zirconia ceramics colorized with the 5 kinds of colorants has no cytotoxicity.
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Objective: To purify the lentinan from sporophore of Lentinus edodes and to study the structure and antitumor activity in vitro of lentinan. Methods: A refined lentinan, named LNT2, was isolated from the sporophore of L. edodes by alkali extraction and alcohol precipitation, and further purified by hydrogen peroxide decoloration and ultrafiltration. The molecular weight and sugar content of LNT2 were measured by HPLC and phenol-sulfuric acid method, respectively. UV was used to detect the protein and nucleic acid in LNT2, and specific rotation was detected. The chemical structure of LNT2 was determined by acid hydrolysis, periodate oxidation, methylation analysis, Fourier infrared spectrum, and NMR experiments. The chain conformation of LNT2 was evaluated by Congo-red test. The inhibitory effect of LNT2 on H22 tumor cells growth was evaluated using MTT assay. Results: The molecular weight, sugar content, and [α]D20 value of LNT2 were estimated to be 185 200, 94.99%, and +8.03°, respectively. UV spectrum showed that there were no peaks at 280 and 260 nm. Chemical and spectroscopic analyses illustrated that LNT2 had a backbone chain of β-(1→3)-linked glucopyranosyl residues and had branches of single glucosyl stubs at C-6 of terminal glucose sugar residues. Congo-red test revealed that LNT2 exhibited a triple helix comformation in low concentration of NaOH solution. Results of the antitumor activity in vitro demonstrated that LNT2 could exhibit the strong effect against the growth of H22 tumor cells and showed a dose-dependent manner. Conclusion: LNT2 is composed of β-(1→3)-linked glucan and has a triple helix comformation. Moreover, LNT2 could present the certain antitumor activities in vitro. Our study lays a solid foundation for the development and utilization of LNT2.
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This study was aimed to explore differential effects of various sections of the velvet antler on promoting cell proliferation in vitro. The NRK-49F cell line from rat kidney fibroblasts was used as the cell model. The cell proliferation rates of the water extracts from the upper, middle and lower section of fresh velvet antler were measured by the MTT method. BCA method was used in the detection of protein concentration. The SDS-PAGE method was used in the analysis of difference composition of the sample protein. The results showed that soluble protein content of the upper, middle and lower section were 17.89, 16.04 and 6.89 mg·mL-1, respectively. From the top to the base, the soluble protein content of velvet antler was decreased. After 24 h treatment, when the protein concentration of the upper and middle section samples of the velvet antler were 800 μg·mL-1 and 600 μg·mL-1, the cell proliferation promoting rates reached the maximum, which were 66.76% and 64.36%, respectively. And when the lower section sample of the velvet antler was 1 000 μg·mL-1, the cell proliferation promoting rates reached the maximum, which was 58.87%. After 48 h treatment, when the upper and middle section samples of the velvet antler were 800μg·mL-1, the cell proliferation promoting rates reached the maximum, which were 219.56% and 215.86%, respectively. And when the lower section sample of the velvet antler was 1 000 μg·mL-1, the cell proliferation promoting rates reached the maximum, which was 169.20%. The velvet antler on the proliferation of cells was much better than the 10% fetal bovine serum. The figure of SDS-PAGE showed the slight difference in the protein composition of three part of the velvet antler. It was concluded that all samples had promoting effects on cell proliferation with concentration-depen-dent, and the main protein in different part of the velvet antler had minor differences.
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Objective: To explore the effects of different pulverization fineness of solid preparations of Chinese materia medica on their in vitro dissolution, using MTT method combined with HPLC. Methods: Compound Biejia Ruangan Tablet (CBRT) was used as model drug, and MTT method was used to obtain the characteristic cell inhibitory rate by different pulverization fineness of dissolving solutions in the dissolution medium (phosphate buffer, pH value 7.4) at different time points. From these results, the cumulative dissolution of CBRT based on cell inhibitory rate was obtained. The dissolution rates of paeoniflorin was determined by HPLC method. Using f2 similar factor, the relevance of these two methods was evaluated. Results: Dissolution of paeoniflorin is changed with the change of pulverization fineness, the accumulative dissolutions of 200 and 300 meshes in vitro were higher than those of other meshes. The results showed that f2 values of 200 and 300 meshes were more than 50, indicating that there was a good correlation between the two methods of measuring the dissolution rate. Conclusion: The results show that the biological potency detection could be used to screen the particle size of CBRT. Considering the production cost of CBRT, 200 mesh is the best particle size.
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OBJECTIVE: To synthesize water-soluble multi-walled carbon nanotubes-polyethyleneimine (MWCNTs-PEI) composite with low cytotoxicity and to lay a foundation for further research of loading drugs with carbon nanotubes. METHODS: MWCNTs-PEI composite was prepared by modifying carboxylated multi-walled carbon nanotubes(MWCNTs-COOH) with polyethylene (PEI), and then the composite was characterized by transmission electron microscopy, infrared spectra, UV spectra and thermalgravimetric analysis. The cytotoxicity of the composite on PC12 cells was measured by methyl thiazolyl tetrazolium(MTT) assay to preliminarily evaluate its biocompatibility. RESULTS: The dissolubilities of MWCNTs-PEI and MWCNTs-COOH complexes were respectively 1.009 and 0.0601 mg · mL-1, and the former was about 16 times of the latter. The cytotoxicity of MWCNTs-PEI composite on PC 12 cells was significantly milder than that of MWCNTs-COOH composite as indicated by MTT assay (P < 0.05). CONCLUSION: MWCNTs-PEI composite not only improves the dispersibility of carbon nanotubes, but also reduces its in vitro cell toxicity.
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Porphyrin-fulleren-based nanoparticles (NP), containing magnetic isotopes 25Mg, 67Zn and zinc of natural isotope composition (Zntotal-NP), have been tested on leukemic cells of patients with T-ALL, B-ALL, AML and lymphocytes of healthy donors. Reliable differences in action of magnetic and non-magnetic zinc isotopes for some types of cells were obtained. Magnetic magnesium isotopes and pure nanoparticles of porphyrinfulleren did not demonstrate any effects. 67Zn-NP induced high cytotoxicity in cells of acute B-lymphoblastic leukemia with LD50 almost three times lower, than those for healthy donors, and 4 times lower in comparison with Zntotal-NP. Also evaluation of apoptosis process in granulocytes of healthy donors in the case of the preparates were performed by method of flow cytometry.
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Objective: To observe the protective effects of Gastrodine on the cultivated rat brain microvessel endothelial cells damage by mimic cerebral ischemia.Methods: The endothelial cells activity,survival rate,the change in NO content,and the effects of Gastrodine were observed in the cultivated rat brain microvessel endothelial cells(BMEC) damaged by mimic cerebral ischemia.Results: The activity and survival rate of BMEC in the ischemia groups are obviously lower than that in the normal groups;compared with the normal groups,the activity,survival rate and NO content of BMEC in the Gastrodin groups have the increasing tendency;comparing to the ischemia groups,the activity of BMEC in the Gastrodin groups obviously increasing(P
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Objective To search for a more efficient method to induce bone marrow stromal cells (BMSCs) into Schwann-liked cells (SLCs) in vitro. Methods On the base of the method which was used by Dezawa (it was looked as the traditional inducing method),some steps were modified,namely,to use all the reagents in half dose in the same time and two times intervally,this method was looked as the modified inducing method.After induced by the two methods,the cells morphologic characteristic,the positive ratio of immunocytochemical dye with anti-S-100 and anti-GFAP,the cells activity measured by MTT method and the DNA percentage in S period measured by flow cytometry were compared with each other respectively to evaluate the methods'effects. Results Compared with the traditional method,the cells induced by the modified method were more similar to the primary Schwann cells in morphology,more positive proportion in immunocytochemic dye with anti-S-100 and anti-GFAP,less damage in the activity and more percentage in S period. Conclusion The modified method had more advantages such as less damage on cells and more efficiency in inducing BMSCs into SLCs.
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Objective:To observe the effects of Chinese herb baicalin on proliferation and cell cycle of periodontal ligament cells(PDLC) by stimulation of lipopolysaccharide(LPS) in vitro.Methods:Human PDLC was cultured in vitro.MTT colormetic method was used to evaluate the effects of Chinese herb baicalin on proliferation of PDLC by stimulation of LPS.Flow cytometry was used to analyze the possible DNA index and cellular cycle changes of PDLC.Results:PDLC showed a significant growth inhibition after being treaded with 100 ?g/ml LPS 3 h by MTT methods,and proliferation ability of PDLC improved greatly after adding baicalin.FCM results showed that the cell number increased in G1 phase,while the cell number decreased in S phase at 12 h after treated with 100 ?g/ml LPS,but the cell number decreased in G1 phase,and increased in S phase after treated with Chinese baicalin.Conclusion:Chinese herb baicalin can significantly promote the proliferation of PDLC.It can make some static cell in G1 phase enter S phrase and improve the growth of human PDLC.
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OBJECTIVE: To observe the inhibiting effect of Interferon-alpha 1b(IFNa-1b) on proliferation of human tenon capsule fibroblasts. METHODS: Human tenon capsule fibroblasts were cultured in vitro and MTT method was used to detect the inhibiting effect of IFNa-1b on human tenon capsule fibroblasts. RESULTS: There was significant difference in OD values between 10~6IU/ml group(0. 1 109?0. 0 585) and control group(0.2535?0. 0502), the inhibition rate in IFN group was 56.25%. CONCLUSION: IFNa-1b has significant effect on inhibiting proliferation of human tenon capsule fibroblasts.
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OBJECTIVE:To study the effects of topirmate on proliferation of neural stem cells in vitro.METHODS:Primary neural stem cells from newborn mice were cultured.The neural stem cells were incubated together with topiramate at 0.4,2 or 10 ?mol?L-1.A vehicle control and positive control were set up,respectively.The quantity and viability of neural stem cells newly generated were analyzed by a convert phase microscope,the neurosphere numbers were counted,and neural stem cells viability was determined and a MTT assay.RESULTS:The number of neurospheres of groups treated with topiramate at 2 and 10 ?mol?L-1 are 20.67?4.04,49.68?3.79,respectively,and the value of MTT assay of the two groups are 0.354 6?0.070 5,0.501 8?0.036 9,it was higher than the vehicle control neural stem cells(7.34?2.31、0.288 0?0.009 3,P
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AIM To investigate the effect of paeonol on proliferation and apoptosis of K 562. METHODS The inhibitory effect of paeonol on K 562 cells proliferation was assayed by MTT dye reduction. HE staining were used to observe the morphology of apoptotic cells, and Flow cytometric analysis were also used to analyze the K 562 apoptosis. RESULTS Pae at the concentration of 7.81～250 mg?L -1 significantly inhibited proliferation of K 562 cells in dose-dependent manner. Typical apoptotic changes were observed in K 562 cells under the light microscope. By FCM methods, the apoptotic rates of K 562 cells were increased from 10.01% to 34.16% after treatment of Pae at concentrations from 7.81 to 125 mg?L -1 for 24 h, which showed obvious concentration-effect relationship. The apoptotic rates of K 562 cells were also increased when Pae (6.25, 25 and 100 mg?L -1) was treated for 6, 12 and 24 h, which showed obvious time-effect relationship. CONCLUSION Paeonol may inhibit K 562 cell proliferation and induce its apoptosis.