Objective:To investigate the feasibility of real-time fluorescence quantitative PCR in the identification of Fritillariae Cirrhosae Bulbus.Methods:The DNA of samples was extracted by magnetic beads ,the primers were amplified by real-time fluores-cence quantitative PCR , and the Cq value and amplification curve were used to determine the samples .Results:The Cq values for five batches of Fritillariae Cirrhosae Bulbus were lower than 30, and the curve had obvious growth period .No Cq value was shown for Fritil-lariae Ussuriensis Bulbus , Fritillariae Thunbergii Bulbus and Bolbostemmatis Rhizoma with straight line curves .Conclusion:The meth-od is simple,feasible and effective in the identification of Bulbus Fritillariae Cirrhosae with high accuracy and good reproducibility .

Objective To investigate the effect of triptolide on asthmatic airway remodeling and signal transducer and activator of transcription 6 (STAT6), acid neutrophil chemokines (eotaxin) impact. Methods The total of 30 mice with ovalbumin (OVA) model of asthma were randomly divided into three groups, control group, asthma group and triptolide group. After 24 hours of the last shot, lung tissue was stained Bronchial inflammatory cell infiltration was determined by using semi-quantitative method and calculate the proportion of goblet cells in airway epithelial cells. Hydroxyproline was determined by McMillan airway mucus score. The mRNA level and protein level of STAT6 and eotaxin in airway epithelium were determined by RT-PCR and immunohistochemistry. Results Compared with asthma group, peribronchial inflammatory cells infiltration of triptolide group were reduced, which mucus index is (1.31 ± 0.23) and hydroxyproline is (284 ± 13) μmg/100 mg. it had a significant in asthma group (P < 0.05). Besides, the protein level and mRNA level of STAT6 and eotaxin were significantly decreased (P < 0.05). Moreover, it was a positive correlation between STAT6 and eotaxin level in airway epithelial (r = 0.668, P < 0.05). Conclusion Triptolide can inhibit airway remodeling and might through the down regulation of STAT6 and eotaxin expression.

Objective To observe the expression of N-terminal pro-brain natriuretic peptide(NT-proBNP),interleukin(IL)-6 and TNF(tumor necrosis factor)-α in patients with acute lung injury(ALI),and to evaluate its value in the evaluation of severity and prognosis of lung injury.Methods A total of 76 patients with ALI treated in this hospital from August 2015 to August 2016 were enrolled in this study and 50 healthy subjects were selected as the control group.The levels of NT-proBNP,IL-6 and TNF-α in the serum of the two groups were compared and their relationship with prognosis were evaluated.Results The levels of NT-proBNP,IL-6 and TNF-α in serum of ALI group were significantly higher than those of healthy control group(P<0.05).The levels of NT-proBNP,IL-6 and TNF-α in the patients with severe lung injury were significantly higher than those in the patients with moderate lung injury(P<0.05);and each index in the patients with moderate lung injury was also significantly higher than those in the patients with mild lung injury(P<0.05).The NT-proBNP,IL-6 and TNF-α levels of the patients with ALI in the death group were significantly different from those in the survival group(P<0.05).Conclusion The levels of serum NT-proBNP,IL-6 and TNF-α can not only reflect the severity of lung injury,but also have a high predictive value for prognosis.

A total of 12 775 SSRs were identified from Scutellaria baicalensis Georgi genomic database, accounting for 2.56% of the total genomic sequences. The result showed that S. baicalensis SSRs were based on 68.32% dinucleotide and 18.63% trinucleotide repeats; CT/GA and TTC/GAA were predominant in the dinucleotide motifs and the trinucleotide motifs respectively. Nine primers were selected to produce highly reproducible SSR bands and were used in studying the genetic diversity of S. baicalensis, 50 individuals from ten populations. 68 SSR polymorphic loci were detected, these loci were polymorphic and displayed 4 to 12 alleles per locus with a mean number of 7; the effect number of alleles was 3. Expected heterozygosities were 0.6 and were far more greater than the average in dicotyledonous plants. PIC (polymorphism information content) was 0.72, Shannon's information index was 1.32, these all proved that S. baicalensis had a high genetic diversity in general. Genetic differentiation among population Gst was 0.131, genetic variation among population accounted for 13.1% and genetic variation within population accounted for 86.9%. The cluster analysis showed that 10 populations S. Baicalensis were classified into 2 groups, but it was not associated with geographical distribution.

The DNA demethylase genes are widespread in plants. Four DNA demethylase genes (LJDME1, LJDME2, LJDME3 and LJDME4) were obtained from transcriptome dataset of Lonicera japonica Thunb by using bioinformatics methods and the proteins' physicochemical properties they encoded were predicted. The phylogenetic tree showed that the four DNA demethylase genes and Arabidopsis thaliana DME had a close relationship. The result of gene expression model showed that four DNA demethylase genes were different between species. The expression levels of LJDME1 and LJDME2 were even more higher in Lonicera japonica var. chinensis than those in L. japonica. LJDME] and LJDME2 maybe regulate the active compounds of L. japonica. This study aims to lay a foundation for further understanding of the function of DNA demethylase genes in L. japonica.

Objective To evaluate the effectiveness of Tanreqing Injection for the preventive of radiation pneumonitis. Methods Retrieving literatures in CNKI, CBM,VIP and WanFang, Medline, Google Scholar database from January 2000 to July 2013.Two researchers extracted and evaluated the quality of all the data independently.RevMan 5.2 software was used for statistical analysis. Results 11 papers and 939 patients were involved in the study. Meta analysis showed statistical difference for the index of marked efficacy between Tanreqing group and radiation group with OR(95%CI)is 0.42(0.30, 0.57). Significant difference was found for the index of more than Grade 3 of radiation pneumonitis, radiation pulmonary fibrosis with OR(95% CI) 0.19(0.11, 0.35), 0.35(0.23, 0.53), respectively(P=0.65). Statistical difference was observed for the index of the dose of radiation and the average onset time of radiation pneumonitis with OR(95%CI) 7.39(4.41, 10.38) and 7.72(6.84, 8.61) respectively. Conclusion Meta analysis based on current limited clinical research results preliminary shows that. Tanreqing injection has a good clinical efficacy for the preventive effect of radiation pneumonitis. The use of Tanreqing injection could reduce the rate of more than Grade 3 of radiation pneumonitis as well as radiation pulmonary fibrosis, extend the onset time and increase the dose which leads to radiation pneumonitis.

Objective: To investigate whether lipopolysaccharide induced parkin expression and mitophagy in macrophages.Methods:The murine peritoneal primary macrophages were aseptically isolated from Kunming mice and cultured in complete medium.The mitochondrial membrane potential of macrophages was detected by flow cytometry,after the cells were stimulated with 200 ng/ml LPS and labeled mitochondria with JC-1.The parkin mRNA level of macrophages was detected by RT-PCR, protein levels of parkin and autophagic related protein LC3 Ⅱ and LC3 Ⅰ were determined by Western blot.The distribution and co-localization of parkin with LC3 and mitochondria in macrophages were respectively observed by laser scanning confocal microscope, before and after the cells were treated with LPS.Results: Flow cytometry results after JC-1 staining showed that mitochondrial membrane potential in macrophages was declined after stimulation with 200 ng/ml LPS, and continuously decreased with prolonged treatment time.The mRNA levels of parkin were increased slightly within 6 h after LPS stimulation,but parkin proteins were increased significantly within 6 h after LPS stimulation.The results of parkin distribution showed that parkin was evenly distributed in the cytoplasm at normal status, but became the obvious punctate distribution after LPS stimulation in macrophages.Western blot results showed LC3 Ⅱ/LC3 Ⅰ levels were increased after LPS stimulation, indicating the appearance of macrophage autophagy.Confocal microscopy showed that there were co-localization of parkin,LC3 and mitochondrial in macrophages after LPS stimulation.Conclusion:Parkin expression is increased significantly and mediated mitochondrial autophagy in macrophages after LPS stimulation, which is involved in the clearance of damaged mitochondria,thereby playing a role in regulating macrophage inflammatory response.

Aim To study the effect of simvastatin on the production of reactive oxygen species ( ROS ) and the secretion of interleukin-1 beta ( IL-1β) in oxidized low density lipoprotein ( oxLDL )-induced macropha-ges. Methods After the murine macrophage J774A. 1 was treated with 0,50,100,200 mg·L-1 oxLDL, the contents of aggregated lipid in macrophages were ob-served and determined by oil red O staining. Then, the oxLDL-primed macrophages were treated with 0 . 5 ,1 . 0μmol·L-1 simvastatin, the production of ROS was de-termined by flow cytometry and the expressions of pro-caspase-1 , cleaved caspase-1 and mature IL-1βon pro-tein level were determined by Western blot. Results The oil red O staining results showed that oxLDL could induce obvious lipid aggregation in macrophages, and reached the saturation point with 100 mg·L-1 concen-tration. Flow cytometry results indicated that oxLDL could induce the production of ROS in macrophages, up to 167% ± 0. 47%, and ROS level decreased to 139% ± 0. 97% in a dose-dependent manner after treatment with simvastatin. Western blot indicated that simvastatin could inhibit the expression of cleaved caspase-1 and mature IL-1β in macrophages triggered by oxLDL;compared with oxLDL group, the expression of cleaved caspase-1 and mature IL-1β decreased in simvastatin treated group, and all results had statistical significance ( P<0. 05 ) . Conclusion In the lipid ag-gregation model of macrophages induced by oxLDL, simvastatin can inhibit the production of ROS, caspase-1 activation, and secretion of IL-1β in macrophages.

Objective To study the effect of macroporous adsorptive resins on the decoloration technology ofLonicera japonica Thunb. polysaccharides(FLP).Methods The effects of 6 kinds of macroporous adsorptive resins i.e. HPD-400A, AB-8, HPD-750, HPD-100, D3520, D301T, S8 on the decolorization technology ofLonicera japonica Thunb. polysaccharides were compared with single factor test in terms of temperature, polysaccharide concentration, pH, adsorption flow, and eluant.Results The decolorization rate and polysaccharide retention rate of S8 were optimal. The best decoloration conditions were as follows: temperature of 40℃, polysaccharide concentration of 5 mg/ml, pH value of 6, flow rate of 1 ml/min, distilled water with pH=6 as eluant. The adsorption rate was 83.2%,and polysaccharide retention rate was 72.1%.Conclusion High decolorization ratio and the high retention rate ofLonicera japonica Thunb. could be obtained by means of decoloration with S8 macroporous adsorptive resins..

AIM: To obtain eukaryotic expression vector of Chinese prostate-specific membrane antigen. METHODS: Chinese prostate-specific membrane antigen (PSMA) cDNA was amplified by RT-PCR from prostate cancer tissues, then cloned into eukaryotic expression vector pcDNA3 0 and sequenced. RESULTS: Seven bases in Chinese PSMA cDNA sequence were found different from those reported by Israeli, which lead to two different amino acids. CONCLUSION: We have obtained the PSMA cDNA, and the recombinant eukaryotic expression vector was successfully constructed. The study lays foundation for DCs vaccine modified by PSMA gene for the treatment of prostate neoplasms.