OBJECTIVE:To optimize the soxhlet extraction technology of Atractylodis macrocephalae,and to provide evi-dence for research and preparation of its formula granules. METHODS:Using the contents of atractylenolide Ⅰ,Ⅱ,Ⅲ as index, based on single factor test,the Soxhlet extraction technology of A. macrocephalae formula granules was optimized and verified by L9(34)orthogonal test with extraction time,solid-liquid ratio,extraction times as factors,and then compared with other technolo-gies (normal temperature extraction method,ultrasonic extraction method,reflux extraction method). RESULTS:The optimal ex-traction technology was as follows as 6-fold ethanol,extracting for 3 times,lasting for 8 h. Results of validation test showed that the extraction amounts of atractylenolide were 0.769,0.752,0.781 mg/g (RSD=1.99%,n=3) for 3 times,which were higher than the extraction amounts of other 3 methods(0.683,0.489,0.693 mg/g). CONCLUSIONS:The optimized extraction technolo-gy possesses high extraction rates of atractylenolide Ⅰ,Ⅱ,Ⅲ,and can be used for the extraction of internal ether from A. macro-cephalae formula granules.

Emotional and cognitive disorders (EACD),such as depression and anxiety,have become very common in today′s society,seriously affecting human lives and health. Neural plasticity can reflect the anti-stress ability of the nervous system to the internal and external stimulation,and is capable of dynamic changes in structure or function to adapt to environmental changes,as is often manifested in the process of compensation and repair of nerve injuries. EACD is often accompanied by macroscopic and cellular morphological changes in brain tissues and functions. Thus,studies on the mechanisms of neural plasticity will contribute to the treatment of EACD. In this paper ,the role of neural plasticity in the active components of Chinese herbal medicine(ACCHM) is reviewed. The effects of ACCHM on 5-hydroxytryptamine(5-HT)system,and the antioxidant activities and neurotrophic effects of ACCHM are described. ACCHM can affect neural plasticity,playing a neuroprotective role by improving 5-HT levels,reducing oxidative stress in brain cells,and increasing the expression of brain derived neurotrophic factor(BDNF). In summary,one ACCHM could affect neural plasticity through one or more mechanisms. There are interactions between different mechanisms of the same ACCHM. Different ACCHM can play a synergistic effect on the enhancement of neural plasticity because of their different mechanisms.

Objective To label rat bone marrow mesenchymal stem cells with superparamagnetic iron oxide (SPIO) and to explore the tropism of BMSCs for hepatocellular carcinoma cells after transplantation in vivo. Methods BMSCs from bone marrow of Sprague-Dawly (SD) rats were cultured isolated and purified, Labeled BMSCs was achieved using Feridex. Twenty-four hepatocellular carcinoma models of SD rats were induced two weeks before tmnsplantation. The models were divided into three groups in random: the labeled BMSCs and unlabeled BMSCs were transplanted respectively into the rat's livers of experimental group (n = 12) and control group A(n =6) via spleens, and no transplant was done for control group B (n =6). MR imaging was performed to monitor the transplanted cells after 1,3,7,14 d using 1.5 T MR system. Signal intensity ratio (SI/SI*) between tumor and hepatic tissue on T_2 * WI were measured and compared by one-factor analysis of variance. After MR imaging, Prussian blue staining was performed. MR imaging findings were compared with histological sections. Results Prussian blue staining confirmed the labeling efficiency of BMSCs was above 90%. SI/SI* of experimental group before and 1, 3, 7, 14 d after transplantation were 3.18±0.21,1.98±0.20,2.38±0.28,2.70±0.25 and 3.16±0.24 respectively. Following transplantation of BMSCs, signal intensity decrease was found in hepatocellular carcinoma of experimental group(F =56.65,P <0.05) and low signal change decreased gradually and disappeared at two weeks after transplantation, while no remarkable low signal change was seen in the control group by T_2 * WI (P > 0.05). A large number of Prussian blue staining positive cells were found in hepatocellular carcinoma in experimental group. Histological section with Prussian blue staining had a good correlation with the signal intensity changes on MR images at different time. Conclusion BMSCs display significant tropism to hepatocellular carcinoma and may be an ideal gene therapy vehicle against hepatocellular carcinoma.

OBJECTIVETo investigate the effect of poly-ADP-ribosylation in hexavalent chromium Cr(VI) induced cell damage.

METHODSThe study object, poly (ADP-ribose) glycohydrolase (PARG) deficient human bronchial epithelial cells (16HBE cells), was constructed previously by our research group. Normal 16HBE cells and PARG-deficient cells were treated with different doses of Cr (VI) for 24 h to compare the differences to Cr (VI) toxicity, meanwhile set up the solvent control group. On this basis, 5.0 µmol/L of Cr (VI) was selected as the exposure dose, after the exposure treatment, total proteins of both cells were extracted for two dimension fluorescence difference gel electrophoresis (2D-DIGE) separation, statistically significant differential protein spots were screened and identified by matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS/MS), and further validated by Western blot.

RESULTSAfter Cr (VI) treatment, the survival rate of PARG-deficient cells was higher than normal 16HBE cells. When the doses reached up to 5.0 µmol/L, the survival rate of 16HBE cells and PARG-deficient cells were respectively (59.67 ± 6.43)% and (82.00 ± 6.25)%, the difference between which was significant (t = -4.32, P < 0.05). 18 protein spots were selected and successfully identified after 2D-DIGE comparison of differential proteins between normal 16HBE cells and PARG-deficient cells before and after exposure. The function of those proteins was involved in the maintenance of cell shape, energy metabolism, DNA damage repair and regulation of gene expression. The differential expression of cofilin-1 was successfully validated by Western blot. The expression level of cofilin-1 in the 16HBE cells increased after Cr (VI) exposure with the relative expression quantity of 1.41 ± 0.04 in treated group and 1.00 ± 0.01 in control group, the difference of which was statistically significant (t = -18.00, P < 0.05), while the expression level in PARG-deficient cells had no statistically significant difference (t = -8.61, P > 0.05).

CONCLUSIONMost of the identified differential proteins are closely related to tumorigenesis, suggesting that poly-ADP-ribosylation reaction may resist the cytotoxicity of Cr(VI) by inhibiting Cr (VI) induced tumorigenesis, which provides important reference data to clarify the mechanisms of poly-ADP-ribosylation in Cr (VI) induced cell damage.

Bronchi ; Cell Transformation, Neoplastic ; genetics ; Chromium ; Cofilin 1 ; DNA Repair ; Epithelial Cells ; Glycoside Hydrolases ; deficiency ; physiology ; Humans ; Tandem Mass Spectrometry

OBJECTIVE： To establish a method for simultaneous determination of 4 triglyceride anti-tumor components in Coix lacryma seed oil. METHODS： HPLC-ELSD was adopted. The determination was performed on Inertsil ODS-3 C18 column with mobile phase consisted of acetonitrile-isopropanol （57 ∶ 43， V/V） at the flow rate of 1.0 mL/min. The column temperature was 30 ℃， and the sample size was 10 μL. Evaporative light scattering detector was used， the drift tube temperature was 70 ℃， and the gas flow rate was 2 L/min. Using glycerol trioleateas internal standard， relative correction factors （RCF） of linolein trilinolein， 1，2-linoleic acid-3-palmitic acid glyceride and 1-palmitic acid-2-oleic acid-3-linoleic acid glyceride were calculated respectively. The contents of above 3 components in C. lacryma seed oil were calculated by RCF. The contents of 4 components in C. lacryma seed oil were determined by external standard. The results of content determination by quatitative analysis of multi-components by single marker （QAMS） were compared with external standard method. RESULTS： The linear ranges were 0.15-4.50 μg for linolein trilinolein， 0.15-4.50 μg for 1，2-linoleic acid-3-palmitate， 0.35-10.50 μg for 1-palmitic acid-2-oleic acid-3-linoleic acid glyceride， 0.35-10.50 μg for glycerol trioleate （r≥0.999 5）. The limits of quantification were 0.13， 0.06， 0.07， 0.12 μg. The limits of detection were 0.04， 0.02， 0.02， 0.03 μg， respectively. RSDs of precision， stability， and repeatability tests were less than 2.0％（n＝6）. The average recoveries were 95.43％-102.67％（RSD＜2.0％， n＝6）. Average RCFs of linolein trilinolein， 1，2- linoleic acid-3-palmitic acid glyceride and 1-palmitic acid-2- oleic acid-3-linoleic acid glyceride were 0.31， 0.88， and 1.21， respectively. RCFs reproducibility was perfect under different experiment conditions. There was no significant difference in results of content determination between QAMS and external standard method （P＞0.05）. CONCLUSIONS： The method is simple， rapid， accurate and reliable. It is used for simultaneous determination of linolein trilinolein， 1， 2-linoleic acid-3-palmitate， 1-palmitic acid-2-oleic acid-3-linoleic acid glyceride and glycerol trioleateas in C. lacryma seed oil.

ObjectiveThe study utilized human transcriptome microarray to explore biomarkers for diagnosing drug-induced liver injury （DILI） caused by anti-tuberculosis drugs. MethodsA 6-month follow-up study was conducted on 152 patients treated with anti-tuberculosis drugs in designated hospitals in Shanghai. The blood samples were collected at the 0， 2， 4， 8， 12 and 24 weeks after treatment. According to the clinical biochemical indicators， the research subjects were divided into DILI cases （34 cases） and Control cases （118 cases）. Single factor analysis was conducted on the influencing factors between the two groups. In a 1∶1 matched DILI-control study， RNA samples of 13 pairs of cases were sequenced by the whole transcript expression mRNA array. Differentially expressed genes （DEGs） were screened by Hotelling's T² value sequencing and the expression trend analysis of genes by STEM （short-time series expression miner）， and the functional enrichment and pathway analysis of DEGs were carried out. ResultsIn total 152 clinical cases， weight of patients was a risk factor for the occurrence of hepatotoxicity caused by anti-tuberculous drugs. Based on the analysis results of mRNA array， 513 DEGs were screened by Hotelling's T² value sequencing method， which were enriched in 32 annotations of GO （Gene Ontology） analysis and 10 pathways of KEGG （Kyoto encyclopedia of genes and genomes） analysis. One differential expression pattern was screened by STEM， which was enriched in 2 biological process notes of GO. Among them， the key genes AIM2， CD86， CXCL10 and non-coding RNAs SCARNA10， SNHG10 and SNORD105 are potential biomarkers of DILI caused by anti-tuberculosis drugs. ConclusionIn this research for biomarkers conducted on cases with liver injury caused by anti-tuberculosis drugs， biological pathways associated with hepatotoxicity are identified and a series of key genes related with drug-induced liver injury are found， which provides the basis for mechanism study and searching for earlier and more sensitive biomarkers.