A method based on gas chromatography-mass spectrometry (GC-MS) was established to analyze the changes of intracellular metabolites and study the toxic mechanisms of different concentrations of particulate matter (PM2.5) effecting the lung tissues in mice.Nasal drip experiments of PM2.5 suspensions (0, 7.5, 20.0, 37.5 g/L) for mice were carried out, and the intracellular metabolites in lung tissues were extracted, pretreated and analyzed.Principal component analysis (PCA) and partial least-squares discriminant analysis (PLS-DA) were employed for pattern recognition, and an obvious distinction among different conditions was found.According to the PLS-DA loading diagram and variable important factor (VIP) value, 7 kinds of potential biomarkers, alanine, valine, leucine, ornithine, fumaric acid, citric acid and purine (p<0.01), were determined with significant differences between four different concentrations of PM2.5.Metabolic pathway analysis indicated that the oxidative stress reactions were enhanced, and the TCA cycle and the purine metabolism in lung cells were restrained after dripping PM2.5 to the lung tissues in mice.This study could provide a new perspective and theoretical basis for the further analysis on toxic mechanisms by PM2.5.

Objective To explore the effect of low calorie metabolism on the survival of HeLa cells exposed to X-rays,and the influence of starvation on the antioxidative factors in the blood of rats after irradiation.Methods MTT method was used to evaluate the impact of different concentration glucose on the proliferation of HeLa cells.Colony formation assay was employed to detect the influence of glucose ( 1,5,10 and 25 mmol/L) on radiosensitivity of HeLa cells. Flow cytometry assay was used to analyze distribution of cell cycle and apoptosis.60 male SD rats were randomly divided into 6 groups with 10 rats each.Rats in every two groups were fed ad libitum,fasted for 24 h and fasted for 48 h,respectively.Rats in one group of each approach were respectively exposed to whole-body X-rays at 11 Gy. At 2 h after irradiation,all of rats were sacrificed and their venous blood was collected.Elisa kits were used to detect superoxide dismutase (SOD) and total antioxidant capacity (T-AOC).Results An increased viability was observed in HeLa cells treated with the glucose at low concentration ( ＜25 mmol/L),while HeLa cell growth was inhibited by glucose at doses of ＞25 mmol/L. Relevant to cells treated with 1 mmoL/L glucose,SERs (sensitive enhancement ratio) in cells exposed to 5,10 and 25 mmol/L glucose were 1.07,1.10 and 1.23,respectively. A reduction of G2/M and S arrests and apeptosis caused by 6 Gy X-ray irradiation were observed [(49.68 ±1.88)％ and (35.54±1.45)％ at G2/M phase,(16.88 ±1.22)％ and (10.23 ±1.65)％ atS phase,t=10.42,5.61,P＜0.05]and in the cells treated with 1 mmol/L glucose compared with cells treated with 25 mmol/L glucose [ ( 25.50 ± 0.95 ) ％ and (7.56 ± 1.07 ) ％,t =21.72,P ＜0.05 ].Without irradiation,calorie restriction exhibited a negligible influence on SOD and T-AOC in rats.However,after 11 Gy irradiation,compared with rats fed ad libitum,the levels of SOD and T-AOC were significantly increased in rats with calorie restriction ( t =40.32,42.78, P ＜ 0.05 ).Conclusions Calorie restriction has a certain radioprotective effect in vivo and in vitro.

The present circuit was designed to apply to human tissue impedance tuning and matching device in ultra-short wave treatment equipment. In order to judge if the optimum status of circuit parameter between energy emitter circuit and accepter circuit is in well syntony, we designed a high frequency envelope detect circuit to coordinate with automatic adjust device of accepter circuit, which would achieve the function of human tissue impedance matching and tuning. Using the sampling coil to receive the signal of amplitude-modulated wave, we compared the voltage signal of envelope detect circuit with electric current of energy emitter circuit. The result of experimental study was that the signal, which was transformed by the envelope detect circuit, was stable and could be recognized by low speed Analog to Digital Converter (ADC) and was proportional to the electric current signal of energy emitter circuit. It could be concluded that the voltage, transformed by envelope detect circuit can mirror the real circuit state of syntony and realize the function of human tissue impedance collecting.
Diathermy ; instrumentation ; Electric Impedance ; Humans ; Radio Waves ; Signal Processing, Computer-Assisted

Objective To study the effect of sanguinarine on growth and radiosensitivity of human glioma A172 cells.Methods MTT assay was used to evaluate cell growth.Cell cycle analysis and reactive oxygen species(ROS) burst were performed by flow cytometry assay.Annexin V/PI assay was used to detect cell apoptosis.Colony formation assay was used to detect the influence of sanguinarine on cell radiosensitivity.Results Exposure of A172 cells to sanguinarine led to dose-and time-dependent cytotoxicity with IC50 values of 4.8,3.9 and 3.2 μmol/L for 12,24 and 48 h,respectively.Furthermore,sanguinarine caused an arrest of S phase.After being treated with 5 μmol/L sanguinarine for 24 h,the ratios of early apoptosis,late apoptosis and necrosis were (60.01 ± 3.73)％,(2.70 ± 0.18)％ and (3.93 ± 0.76)％ respectively in A172 cells compared with the untreated control(t =55.28,8.32,9.51,P ＜0.05).An increased generation of ROS was found after treatment with sanguinarine,however,NAC inhibited the effect of sanguinarine.As analyzed with multi-target click model fitting curves,the SERD0 of sanguinarine-treated cells was 1.48.Conclusions Sanguinarine inhibits the A172 cells growth via apoptosis induction and ROS burst.Moreover,sanguinarine at a non-cytotoxicity dose increases cell sensitivity to X-rays.

This research explored the synergistic effects and the potential mechanisms of RCE-4 and various nonsteroidal anti-inflammatory drugs (NSAIDs) on the proliferation of cervical cancer Ca Ski cells. The MTT assay and CalcuSyn V2.0 software were used to detect cell proliferation and calculate the combination index (CI); the expression levels of various proteins were analyzed using Western blot assay; mitochondrial membrane potential (MMP) was assessed using JC-1 staining; acridine orange/ethidium bromide (AO/EB) double-fluorescence staining was used to detect the apoptosis of Ca Ski cells; a co-immunoprecipitation (Co-IP) assay was used to analyze the relative content of Bcl-2-Beclin 1 complex in Ca Ski cells. The results demonstrate that the combination of RCE-4 and NSAIDs increases the inhibition of Ca Ski cells compared to the single-RCE-4 group, and celecoxib provided the best synergistic effect among the four NSAIDs tested, with a CI of 0.32. The combination of RCE-4 and celecoxib significantly down-regulated the expression of cyclooxygenase-2 (COX-2) and nuclear transcription factor-κB (NF-κB), and promoted the expression of non-steroidal anti-inflammatory drugs activity gene-1 (NAG-1). In addition, autophagy induced by RCE-4 was markedly inhibited in combination with celecoxib, which was associated with down-regulation of the expression of microtubule-associated protein 3 (LC3)-II, Beclin 1, p62 and autophagy-related gene (ATG) 3/4B/5/7/14. RCE-4-induced apoptosis was significantly enhanced by altering the depolarization of mitochondrial membrane potential and the expression of B cell lymphoma-2 (Bcl-2), B cell lymphoma-xl (Bcl-xl), Bcl-2 associated X protein (Bax), Bcl-2/Bcl-xl-associated death promoter (Bad) and cleaved cysteinyl aspartate specific proteinase (cleaved-caspase) 3/7/9. Furthermore, the formation of the Bcl-2-Beclin 1 complex was significantly inhibited in Ca Ski cells treated with RCE-4 in combination with celecoxib. Taken together, this research shows that the combination of RCE-4 and celecoxib has a significant synergistic effect on the proliferation of Ca Ski cells by promoting apoptosis, inhibiting autophagy and disturbing the formation of the Bcl-2-Beclin 1 complex, which may be a novel strategy to increase the sensitivity of anti-cervical cancer drugs.

OBJECTIVE: To optimize the extraction technology of Bushen yangxue granules, and to provide reference for developing hospital preparations. METHODS: The orthogonal test was designed to optimize the extraction technology of Bushen yangxue granules using the comprehensive score including the content of hyperoside and total flavonoids, extract yield as evaluation indexes, water amount, extraction time and extraction times as factors. Verification test was also conducted. RESULTS: The extraction technology of Bushen yangxue granules was 10-fold water, extracting for 2 times and 1 hour each time. The results of verification test indicated that the contents of hyperoside and total flavonoids were 0. 154 mg/g (RSD=0. 010％, n=3) and 11. 094 mg/g (RSD=0. 008％, n=3). The average extract yield was 37. 76％ (RSD=0. 008％, n=3). CONCLUSIONS: The optimized extraction technology is stable and feasible, which can provide the basis for developing hospital preparations of Bushen yangxue granules.

ObjectiveTo explore the prognostic value of pretreated maximum standardized uptake value (SUVmax) using 18-fluorodeoxyglucose positron emission tomography with computed tomography (18FDG PET/CT) in locally-advanced nasopharyngeal carcinoma (NPC) treated by intensity-modulated radiotherapy (IMRT).MethodsOne hundred and forty previously untreated stage Ⅲ - Ⅳb ( UICC/AJCC 6th) patients with biopsy-proven nasopharyngeal carcinoma were examined.All of the enrolled patients accepted whole body/head-neck 18FDG PET/CT before radical IMRT. 18FDG uptakes were recorded as SUVmax of primary tumor (SUVmax-P) and SUVmax of cervical lymph nodes (SUVmax-N).The relationships between SUVmax and long-term clinical outcomes were analyzed.ResultsThe median SUVmax-P was 10.4,and the median SUVmax-N was 6.2.The SUVmax-P was weakly correlated with T-stage ( R =0.279,P =0.001 ).The SUVmax-N was weakly correlated with N stage ( R =0.334,P =0.000 ).There were no difference of the median SUVmax-P (9.2 vs.10.4,U =560.50,P =0.805 ) and the median SUVmax-N (4.0vs.5.0,U =576.00,P =0.908) between patients with and without local recurrence.The median SUVmax-P of patients with distant metastasis was significantly higher than those without metastasis (11.9 vs.9.8,U =987.50,P =0.014).The SUV of 10.2 was taken as a cut-off for high and low uptake tumors.For patients with SUVmax-P ＞ 10.2,the 5-year distant metastasis-free survival (DMFS) and 5-year overall survival (OS)were significantly higher than those with SUVmax-P ≤ 10.2 (69.1％ vs.95.5％,x2 =15.88,P =0.000;68.4％ vs.94.0％,x2 =15.56,P =0.000,respectively).Multivariate analysis showed that SUVmax-P was the only independent risk factor of 5-year DMFS and OS ( HR =7.87,P =0.001 and HR =5.14,P =0.003). Conclusion SUVmax-P is a useful biomarker predicting long-term clinical outcomes in newly diagnosed locally-advanced NPC patients.

The structural theory and fabrication of a self-designed rotating auxiliary spring in edgewise technique were described. Rotating auxiliary spring was applied to correct rotated tooth in clinic. It indicated that rotating auxiliary spring was an easy-fabricated, convenient-manipulated device, and could produce an effective treatment results in correcting rotated tooth, especially in the final adjustment stage.
Humans ; Malocclusion ; Tooth Movement Techniques

AIMTo study the antitumor peptide components in the stems and leaves of mistletoe (Viscum coloratum (Kom.) Nakai), the primary structure of the novel peptide was elucidated.

METHODSCation exchange, gel filtration and HPLC were employed for isolation and purification. Matrix Assisted Laser Desorption Ionization-Time of Flight-Mass Spectrometry was used to determine the mass. The complete amino acid sequence of the novel peptide was obtained by Edman degradation combined with enzyme digestion. The antitumor activity of the peptide in vitro was studied with MTT method.

RESULTSThe primary stucture of the peptide named as viscotoxin B2 is KSCCKNTTGRNIYNTCRFAGGSRERCAKLSGCKIISASTCPSDYPK. The IC50 value of viscotoxin B2 on the Rat Osteoblast-like Sarcoma 17/2.8 cells in vitro is 1.6 mg x L(-1).

CONCLUSIONViscotoxin B2 in V. coloratum, which has high similarity with viscotoxins from V. album, showed antitumor activity.

Amino Acid Sequence ; Animals ; Antineoplastic Agents, Phytogenic ; chemistry ; isolation & purification ; pharmacology ; Bone Neoplasms ; pathology ; Inhibitory Concentration 50 ; Molecular Sequence Data ; Molecular Weight ; Osteosarcoma ; pathology ; Peptides ; chemistry ; isolation & purification ; pharmacology ; Plant Leaves ; chemistry ; Plant Proteins ; chemistry ; isolation & purification ; pharmacology ; Plant Stems ; chemistry ; Plants, Medicinal ; chemistry ; Rats ; Tumor Cells, Cultured ; drug effects ; Viscum ; chemistry

OBJECTIVETo investigate the effect of AMPK agonist 5-aminoimidazole-4-carboxamide ribonucleoside (AICAR) on proliferation, differentiation and apoptosis of U937 cells and explore its possible mechanism.

METHODSU937 cells were cultured with different concentrations of AICAR for 24 h and 48 h. Cell proliferation was evaluated. Cell growth curve was analyzed by CCK-8; cell apoptosis was analyzed by cell morphology, Annexin V/7-AAD double labeling. The differentiation of U937 cells was evaluated by expression of CD11b. The Bcl-xL, Bax, Bim, caspase-3 mRNA expressions of U937 cells were determined by real time PCR.

RESULTSAICAR significantly inhibited the growth of U937 cells in a time-and dose-dependent manner, with a 24 h IC50 value of 1.1 mmol/L and 48 h of 0.9 mmol/L. 1.0 mmol/L AICAR didn't induce differentiation of U937 cells with the increase of CD11b expression for 24 h (P > 0.05). The U937 cells apoptosis was confirmed by cell morphology and Annexin V/7-AAD labeling. AICAR induced apoptosis of U937 cells and the apoptosis rate was (6.81 ± 1.16)% at 1 mmol/L AICAR higher than control group (2.74 ± 0.32)% without AICAR for 24 h treatment (P < 0.05). The real time PCR assay revealed that as compared with control group, the expression of Bim and caspase-3 mRNA were increased, while Bcl-xL and Bax were unchanged on the AICAR treatment.

CONCLUSIONAICAR can effectively inhibit proliferation and induce apoptosis of U937 cells. However, it has no significant effect on differentiation of U937 cells. The mechanism may be related with up-regulating Bim and Caspase-3.

Aminoimidazole Carboxamide ; analogs & derivatives ; pharmacology ; Apoptosis ; drug effects ; Cell Differentiation ; drug effects ; Cell Proliferation ; drug effects ; Humans ; Ribonucleotides ; pharmacology ; U937 Cells