AIM:To establish the chromatography fingerprint of the constituents of Melia toosendan Sieb from different areas in order to provide a base for the identification of its quality. METHODS: A gradient separated method was applied. Column:Inertsil ODS-3 C_ 18 ,mobile phase:acetonitrile-water,detection wavelength:270 nm,flow rate:1.0 mL/min,column temperature:25 ℃. RESULTS: To establish the chromatography fingerprint of the constituents of Melia toosendan Sieb., make the technical parameters for its quality controlling,and mark 39 main peaks as its characteristic fingerprint. CONCLUSION: The distribution of constituents of Melia toosendan Sieb differ a little from different areas, but the propotion of the constituents differ greatly, with the Melia toosendan Sieb.from the same area , the distribution and propotion differ a little. This method is reproducible, simple and easy, and can be use to provide a base for the quality control of Melia toosendan Sieb.

A sensitive and reliable liquid chromatography-electrospray ionisation-tandem mass spectrometry (LC-ESI-MS/MS) method was established to simultaneously quantitate four categories of compounds (isoflavonoids, flavonoids, alkaloids and saponins) in Gegen-Qinlian decoction (GQD). These compounds were separated by a Shiseido CAPCELL PAK C18 column with a linear gradient consisting of 0.1% (v/v) formic acid in water (A) and 0.1% (v/v) formic acid in acetonitrile (B), and delivered at a flow rate of 0.3 mL/min. All the analytes were determined by electrospray positive ionization tandem mass spectrometry in a multiple reaction monitoring (MRM) mode. Linearity, accuracy, precision, recovery and stability of the method were evaluated with the validation over the range of 4.0-538 5 ng/mL. The proposed method was applied to the analysis of a Chinese herbal preparation GQD successfully.

N 6-methyladenosine (m 6A) is the most abundant RNA base modification in mammals, especially in eukaryotic messenger RNA (mRNA). N 6-methyladenosine modification can regulate RNA splicing, translocation, stability and ultimately affect protein synthesis. m 6A modification is catalyzed by RNA writers, reduced by erasers and also be recognized by readers. Abnormal changes ofm 6A levels are closely related to tumor occurrence and development, including proliferation, growth, invasion and metastasis. In the process of tumor radiotherapy, m 6A modification affects the efficacy of radiotherapy by affecting DNA damage, tumor stem cell generation and tumor cell radiation sensitivity. This article reviews the role of m 6A-modified epigenetic regulation in malignant tumors and the research progress of its mechanism in tumor radiotherapy, in order to provide new ideas for the development of clinical tumor molecular targeted therapies and radiosensitizers.

Objective To study the detection rate of early esophageal carcinoma using magnifying endoscopy,and to evaluate the relationship between the imaging patterns of microvasculature change and his- tological diagnosis.Methods Two hundred and fourteen patients with esophageal mucosa roughness,ero- sion,plaque,abnormal color and indentation in conventional endoscopy and 16 healthy volunteers were en- rolled.The magnifying endoscopy images were graded as four patterns by intraepithelial papillary capillary loop(IPCL)changes after iodine dyeing.The biopsies underwent pathologic evaluation.The comparison be- tween the imaging patterns of endoscopy and histological diagnosiswas was evaluated.Results 80.4%(90/ 112)esophagitis was type 2,and 85.7%(12/14)early esophageal carcinoma was type 3 and type 4.The difference was significant between early esophageal carcinoma and normal mueosa(?~2=27.32,P

Objective Through the investigating the molecular expressions of integrin?_5?_1,Fn and CD_(44v6) in ovarian epithelial neoplasms,this study is trying to explore the relationship between the lymphatic spread of the tumor with these molecules.Methods The expression of 80 cases of integrin?_5?_1,Fn and CD_(44v6) was examined through ElivisionTM immunohistochemistry method in ovarian epithelial neoplasm.The system of image analysis was used to measure the expression of various molecules quantitatively.Results Qualitative and semi-quantitative results:The expression levels of integrin?_5?_1 and Fn in the ovarian epithe- lial neoplasms had downward tendency in the order of benign,boundary and malignant neoplasms and there was a significant difference in the expression levels of integrin?_5?_1 and Fn(P

Aim The effect of capsule bushenyanshou (BSYS), a compound of traditional Chinesemedicine, on the immunopharmacological activities of mice was investigated. MethodsThe indexes of immunopharmacological activity, such as the clearance rate of charcoalparticles, the lymphocyte transformation and the content of serum hemolysin, were mea-sured. Results Capsule BSYS (400, 800 mg?kg-1, qd ? 12) markedly increased theclearance rate of iv charcoal particles and 1he lymphocyte transformation stimulated invivo by PHA in mice. In hydrocortisone -treated mice(15 mg? kg-1, sc, qd ? 5), capsuleBSYS significantly enhanced the content of serum hemolysin and the weights of spleenand thymus. The results also demonstrated capsule BSYS performed a sighted inhibition ofthe delayed type hypersensitivity in mice. Conclusion Capsule BSYS has the capacity ofimmunological intensification and regulation.

ObjectiveTo investigate the effect of immunoglobulin G (IgG) extravasated from blood circulation on the expression of toll-like receptor 4 (TLR4) induced by peripheral lipopolysaccharide (LPS) in rat brain. Methods The rats were divided into four groups in random, 5 rats in each. Group one received LPS 100μg/kg by intraperitoneal administration, normal saline was given by intravenous injection 6 hours later; group two was injected with adrenalin (AD) 15μg/kg intravenously; group three was treated with LPS intraperitoneally, AD was injected 6 hours later; group four was injected normal saline intravenously as control. For all groups, the animals were sacrificed 30 min after the last injection, and the brains were taken for investigation of the TLR4 expressions by immunofluorescence staining and RT-PCR. Result Immunofluorescence staining showed that IgG immunoreactive product was patch-like, distributed in the brain parenchyma in all the animals that received AD. In the LPS+normal saline group, IgG was found merely around the blood vessels. Meanwhile, in LPS+AD animals, TLR4 immunoreactive product coexisted with microglia marker Iba-1 within the IgG extravasated area. The double-labeled cells dispersed in the brain parenchyma and near to the cerebral vessels. In the LPS+saline group, TLR4 positive cells were endothelial-like. RT-PCR results indicated that the expression level of TLR4 in the LPS+AD group were significantly higher than that in the LPS+saline group or AD group or the saline control (P<0.01). Conclusion Extravasated circulating IgG may enhance the TLR4 expression in the rat brain induced by peripheral LPS.

AIM To investigate whether pioglitazone can protect cortical neurons from lipopolysaccharides(LPS)-induced neurotoxicity and the mechanisms responsible for this protective effect. METHODS After 7 d cultures，cultured cortical neurons were incubated with LPS 10 mg·L~(-1) for 4-24 h with or without other drugs. In co-incubation experiments, other drugs were added to the neurons 30 min or 1 h prior to incubation with LPS. The cell viability was assessed by MTT assay. The neuronal apoptosis was quantified by scoring the percentage of cells with apoptotic nuclear morphology after Hoechst 33258 staining. The cultured cells were then fixed on the 7th day and immunocytochemically stained with phosphorylated JNK1 antibody. The protein expressions of active caspase 3 and phosphorylated JNK1 were measured by Western blot. Nitric oxide (NO) generation was measured by Griess method. RESULTS The decrease of cell viability and the increase of apoptotic cells in cultured cortical neurons were observed incubated with LPS for 24 h compared with the normal controls. The cell viability of cortical neurons was decreased from (100.0±10.9)% in the normal control group to (72.3±2.1)% in the LPS-treated group and the apoptotic cell percentages were increased from (11.5±4.2)% in the normal control group to (39.5±8.2)% in the LPS group. LPS induced the increases in phospho-JNK1, active caspase 3 expression, and NO generation. Pioglitazone 0.01, 0.1 and 1 μmol·L~(-1), respectively inhibited LPS-induced decrease in cell viability and increase of apoptotic morphology, active caspase 3 expression in cultured neurons. In LPS+pioglitazone 1 μmol·L~(-1) group, cell viability was （97.8±9.7）%, the apoptotic cells percentage was (20.6±5.0)%, NO generation (6.8±1.3)μmol·L~(-1). Furthermore, pioglitazone also inhibited LPS-induced the increase in JNK1 phosphorylation and NO generation. JNK inhibitor SP600125 5 μmol·L~(-1) significantly inhibited LPS-induced neurotoxicity, cell viability was increased from (72.3±2.1)% to (109.8±11.8)%, the apoptotic cells percentage from (39.5±8.2)% decreased to (19.1±4.8)%, NO generation from （21.1±5.0）μmol·L~(-1) decreased to（4.0±1.3）μmol·L~(-1). The PPARγ antagonist GW9662 10 μmol·L~(-1) did not reverse the effects of pioglitazone. In LPS+pioglitazone 1 μmol·L~(-1)+GW9662 10 μmol·L~(-1) group, cell viability was (90.7±6.9)%, the apoptotic cells percentage was (23.4±4.1)%, and NO concentration was (5.8±0.7)μmol·L~(-1). CONCLUSION Pioglitazone protects cortical neurons against LPS insult at least via inhibiting JNK activity and NO generation, but not PPARγ activation.

Bronchi ; drug effects ; pathology ; Cell Line ; Cell Transformation, Neoplastic ; drug effects ; genetics ; Dust ; Epithelial Cells ; drug effects ; pathology ; Gene Library ; Humans ; Minerals ; toxicity

Stimulator of interferon genes (STING) is an important protein of the innate immune response, and protects against viral infections. To search for an alternative splicing isoform of STING, we undertook rapid amplification of cDNA ends (RACE) and RT-PCR with RNA extracted from human embryonic kidney (HEK) 293 cells and primers designed according to the mRNA sequence of full-length STING(NM-198282. 82). The new sequence was compared using a bioinformatics method. Then, a newly discovered, alternative splicing isoform of STING, named "STING(sv)", and STING(wt) were subcloned into the eukaryotic expression vector pEGFP-C1 and pcDNA 3. 1. Whole-cell extracts were analyzed by western blotting and then probed with monoclonal antibody against enhanced green fluorescent protein (EGFP) after transfection of EGFP-STING(wt) and EGFP-STING(wt) plasmids in HEK293 cells. pcDNA-STING(wt) and pcDNA-STING(wt) were transfected in HEK293 cells, and the luciferase assay carried out. Compared with STING(wt), STING(sv) lacks exon 7 so that shift in the reading frame may produce a protein with a different C-terminal in amino acids 1-30. Western blotting confirmed an expected strong band at 58 x 10(3) kD. The functional luciferase assay showed that STING(sv) inhibited the activity of the interferon (IFN)-β promoter. STING(sv) can be expressed in multiple tissues and distinct cell lines. Our discovery of a new, alternative splicing isoform of STING provides new insights into the functional regulation of STING. STING(sv) could be a dominant negative inhibitor for the activity of the IFN-β promoter in the virus-infection pathway. Hence, STING(sv) could participate in the "fine tuning" of the virus-induced activation of IFN. Therefore, exploring the role of STING(sv) in the pathogenesis of human diseases could be very worthwhile.
Alternative Splicing ; Amino Acid Sequence ; HEK293 Cells ; Humans ; Interferon-beta ; genetics ; Membrane Proteins ; genetics ; metabolism ; Molecular Sequence Data ; Promoter Regions, Genetic ; Protein Isoforms ; genetics ; metabolism ; Sequence Alignment