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.

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.

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

Objective To study the clinical application of the lung lesion biopsy through helical CT-guided fine needle aspiration. Methods 44 cases of lung lesions were pathologically biopsy diagnosed through helical CT-guided fine needle aspiration. Results All 44 lung lesions were successfully conducted biopsy through CT-guided fine needle aspiration, and the pathological confirmed diagnosis rate was 98%. Conclusion With high diagnosis rate, CT- fine needle aspiration is clinically applicable and the complication is rare. It is easily-operated, safe and economical.

Objective To explore the difference in pain threshold between related acupoints and the specificity of acupoints in cholecystitis patients.Method Actual measurement was made in 80cases, a normal group of 30 cases (volunteers without psychological and physiological diseases) and a patient group of 50 cases (volunteers with cholecystitis). The probe of an algometer was perpendicularly placed on the selected acupoint, the pressure point was pushed and pressing was stopped immediately after the subject felt pain. The pain thresholds displayed by the measured acupoints were recorded. The pain thresholds of the selected acupoints were compared between the normal and patient groups and thedifferences were analyzed to explore the specificity of acupoints.Result A comparison of the pain thresholds of the Back-Shu points of the Bladder Meridian of Foot-Taiyang between the patient and normal groups showed that there were no significant differences in the thresholds of the left Back-Shu points Geshu (BL17), Ganshu (BL18), Danshu (BL19), Pishu (BL20), Weishu (BL21) and Dachangshu (BL25) of the bladder meridian between the two groups (P>0.05) and there were significant differences in the thresholds of the right Back-Shu points Geshu, Ganshu, Danshu, Pishu, Weishu and Dachangshu of the bladder meridian between the two groups (P<0.01). The pain threshold was significantly lower in thepatient group than in the normal group. A comparison of the pain thresholds of the main points of the Liver Meridian of Foot-Jueyin, the Gallbladder Meridian of Foot-Shaoyang and the stomach meridian between thepatient and normal groups showed that there were significant differences in the thresholds of the right points Liangmen (ST21), Riyue (GB24) and Qimen (LR14) of the liver, gallbladder and stomach meridians (P<0.01) and no significant difference in the threshold of the right point Taichong (LR3) between the two groups (P>0.05). The pain thresholdwas significantly lower in thepatient group than in the normal group. There were no significant differences in thethresholds of the left points Liangmen, Riyue, Qimen and Taichong of the Liver Meridian of Foot-Jueyin, the Gallbladder Meridian of Foot-Shaoyang and the stomach meridian between the patient and normal groups (P>0.05).Conclusion The pain sensitivity of ipsilaterali related acupoints increase and the relative specificity of acupoints exist in cholecystitis patients.

Objective To study the process of foam fractionation of polyphyllin in semi-batch mode. Methods Taking enrichment ratio, recovery rate of polyphyllin, and purity ratio as the performance criteria and using single examining method to examine the operational parameters, i.e. operation mode, air flow rate, initial feed concentration, solution pH value, initial feed height and temperature on separation performance. The optimal conditions of the process were obtained finally. Results The separation performance is good when gas flow rate is 400 mL/min, initial feed concentration (polyphyllins content) is 0.3 mg/mL, pH value is 7, feed height is 30 cm, and feed temperature is 40 ℃. The enrichment ratio is 25.7, recovery ratio is 42.1%, and the foam liquids purity of total polyphyllin is 41.4%, which is 4.5 times higher than that in feed purity. Conclusion Foam fractionation technique could be applied to separate polyphyllin.

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

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.