This study was aimed to screen candidate genes involved in the triterpenoid saponins biosynthetic pathway of the Ilex asprella root. The Illumina platform was applied to perform transcriptomic sequencing of I. asprella root, followed by a series of bioinformatics analysis. The results showed that a total of 272 candidate unigenes were anno-tated to be involved in the biosynthetic pathway of terpenoid in the transcriptome of I. asprella root, including 72 u-nigenes for the upstream pathway and 26 unigenes for cyclization, oxidation and glycosylation in the downstream pathway. Phylogenetic analysis was carried out to further analyze the evolution relationship of some candidate uni-genes and their homologous genes. Two genes IaA S1 and IaA S2 were proved to be mixed amyrin synthases in yeast expression system. Moreover, IaA S1 was identified to one of the rare ASs with α-amyrin as the major product. It was concluded that a series of candidate genes, which might be involved in the biosynthetic pathway of triterpenoid saponins, were screened out from the transcriptome of I. asprella root. Further investigation of these candidate genes will provide insight into their actual functions in the triterpenoid saponins biosynthetic pathway in I. asprella.

Objective To evaluate the inhibitory activity of Herba Taraxaci extract on Escherichia coli DH5α (E. coli DH5α) and to investigate proteomic response of E. coli. Methods Medicinal powder of Herba Taraxaci was extracted with the solvents of different polarity ( n-hexane, ethyl acetate, distilled water) , and then the obtained 8 different extracts were subjected to thin layer chromatography ( TLC) analysis. Microdilution method was performed to detect the minimum inhibitory concentration ( MIC) of different extracts and the growth curves were described. The protein expression profiles of E . coli treated with the extracts were analyzed by sodium dodecyl sulfate polyacrylamide gel electropheresis ( SDS-PAGE) and two dimensional electrophoresis (2-DE) . Results Water decoction of Herba Taraxaci could obviously suppress the growth of E. coli with a MIC of 1.95 mg/mL. The different extractions exhibited no antibacterial activity except ethyl acetate phase 3 with a MIC of 0.13 mg/mL, which was equal to 19.23 mg/mL of crude drugs. The results of TLC analysis showed that chlorogenic acid was undetectable in n-hexane extract and ethyl acetate phase 1 extract, and ethyl acetate phase 2 and 3 extracts showed obviously increased spots. The results of SDS-PAGE and 2-DE showed that water decoction of Herba Taraxaci had inhibitory effect on the expression of functional protein. The results of 2-DE showed that after treatment with ethyl acetate phase 3 at the concentration of 2 × MIC for 21 hours, the amount of protein spots were 92 less than those of the blank control group, the spots of E. coli DH5α soluble protein with expression amount down-regulated doubly were 24, and those with expression amount up-regulated doubly were 19. Ethyl acetate phase 3 extract had an effect on down-regulating the protein expression of E. coli DH5α soluble protein pH3-10, and water decoction of Herba Taraxaci had inhibitory effect on E. coli DH5αprotein expression. Conclusion Herba Taraxaci has significant antibacterial activity on E. coli DH5α, and the water-soluble fraction of chlorogenic acid and caffeic acid might be the active components. The possible antibacterial mechanism may be related with the regulation of bacterial protein expression.

Objective To establish a molecular identification method for three cultivars of Amomum villosum Lour.(AVL),thus to provide scientific evidence for the identification,selection and breeding of AVL.Methods The fragments of 26S rDNA D1-D3 region and matK gene of three cultivars of AVL and Amomum longiligulare T.L.Wu were amplified by polymerase chain reaction(PCR) and with corresponding primers,and then their sequences were analyzed,and phylogenetic tree was constructed based on the sequences.Results We obtained 739 bp in 26S rDNA D1-D3 sequence.Differences in 4 basic sites of 739 bp were shown between AVL and Amomum longiligulare T.L.Wu.The two cultivars of AVL,Changguo and Yuanguo,had the same sequence,but there was a difference in one basic site of Changguo and Yuanguo from Chunxuan.The phylogenetic tree based on 26S rDNA D1-D3 sequence revealed the difference between Chunxuan and the other two cultivars of AVL.We also obtained 824 bp in matK gene sequence.The three cultivars of AVL showed the consistent sequence,but there was a difference in one basic site of three cultivars of AVL from Amomum longiligulare T.L.Wu.Conclusion We can identify the three cultivars of AVL through the sequence differences at the molecular level,and Chunxuan has a closer genetic relationship with Amomum longiligulare T.L.Wu.

Objective To establish the quality standard of Radix Toddaliae Asiaticae. Methods Thin layer chromatography ( TLC) and high performance liquid chromatography ( HPLC) were used to identify and determine chloride nitidine and toddalolactone in Radix Toddaliae Asiaticae. The moisture and total ash contents were detected according to the methods recorded in appendix of Chinese Pharmacopeia (2010 edition) . Results Toddalolactone and chloride nitidine were detectable by TLC, the spots were clear and the dissociation was good. The established HPLC method was simple and accurate. The linear ranges of toddalolactone and chloride nitidine in Radix Toddaliae Asiaticae were 2.84~42.6 μg/mL and 25.6~385 μg/mL, and their recovery rates were 99.2 % ( RSD=1.12%) and 100 % ( RSD=0.71%) , respectively. The content of moisture was in the range of 75.8~98.9 mg/g and that of total ash was in the range of 12.4~33.6 mg/g. Conclusion The developed method is specific and accurate, and can provide useful reference for establishing quality standard of Radix Toddaliae Asiaticae.

Biological chemistry of Chinese herbal germplasm resources (BCCHGR) is a new cross-discipline formed from rapid development of modern science and technology and its application in the area of Chinese herbal resources. BCCHGR was defined as probing and understanding biological processes like heredity, gene transcription, expression and metabolism of Chinese herbal germplasm, at the interface of biochemistry, molecular biology and chemistry, elu-cidating the nature of Chinese herbal germplasm using as TCM medicine as well as the forming mechanism thereof. In this paper, the scientific background, definition, significance and contents of BCCHGR were discussed to depict a preliminary picture of BCCHGR and arouse popular consideration and discussions.

Objective To obtain a more accurate assessment of prognostic significance of NLR in ovarian cancer. Methods A systematic literature search was conducted using the electronic databases PubMed ,Web of Science,and Embase up to May 2016. Hazard ratio(HR)and odds ratio(OR)with 95% confidence interval (95%CI)were calculated. Statistical analysis was performed using Stata 12.0. Results 12 studies,consisting of 3 ,854 patients ,were selected in this meta-analysis. High NLR level was significantly associated with poorer overall survival(OS)(HR:1.69)and shorter progression free survival(PFS)(HR 1.63). Additionally,increased NLR was significantly correlated with advanced FIGO stage(OR 2.32),higher level of CA-125(OR 3.33),more extensive ascites(OR 3.54)as well as less chemotheraputic response(OR 0.53). Conclusions Elevated pretreat-ment NLR can serve as a predicative factor of poor prognosis for ovarian cancer.

Objective An investigation of new cultivars of Amomum villosum Lour.(AVL) with high yield and good quality was carried out,thus to supply evidences for the identification of AVL cultivars in accordance with the morphological features of their flowers and fruits.Methods An investigation of AVL species from the genuine producing areas of Yangchun city of Guangdong province was performed.The morphological features of flowers and fruits of two cultivars(Changguo and Yuanguo) as well as one breeding type(Chunxuan type) were examined.Results Specific and significant features were screened out in different cultivars of AVL.Conclusion There exit specific features in flowers and fruits of different cultivars of AVL from Yangchun.

Objective To establish a DNA extraction method for DNA barcoding analysis of Chinese medicinal materials.Methods Seven different DNA extraction methods were used to extract DNA from 6 medicinal recalcitrant plants which are rich in secondary metabolites.Results CTAB method 3 was fast,simple,universal and effective,by which a high DNA concentration and qualified ratio were obtained as compared with the other methods.The DNA extracted by this method could provide good results for DNA barcoding analysis.The main improved steps of this methods were as follows:①adoption of 3 %CTAB rather than 2 %CTAB in the exaction;②adding 1 %polyvinylpyrrolidone(PVP) and 0.2 %?-mercaptoethnoal in extraction solution to remove secondary metabolites and to prevent DNA degradation;③centrifuge at 10000 r/min for 15 min to remove protein and impurity.Conclusion CTAB method 3 is a proper method of DNA extraction for DNA barcoding analysis of Chinese medicinal materials.

Objective The aim of this study was to investigate the association between maternal serum lev-el of neutrophil-lymphocyte ratio(NLR)and preterm labor in late stage of pregnancy. Methods In this retrospec-tive analysis of 200 pregnant women who underwent routine prenatal examination and delivered from January 12016 to December 312016 in the Maternal and Child Health family planning services,Xinhui District,Jiangmen city. The patients were divided into the PIR group(n = 100)and the control group(n = 100). The levels of leuko-cytes,neutrophils,lymphocytes,CRP and NLR of the two groups were compared. Multivariate analysis was per-formed to identify independent risk factor. Logistic regression model was established and the area under the receiver operating characteristic curve(ROC)was recorded for analysis of the prognostic value of different serum inflamma-tory makers in late stage of pregnancy. Results The NLR predict premature occurrence of AUC,the sensitivity and specificity of which were higher than those of peripheral blood leukocyte,neutrophil count,lymphocyte count and CRP. Conclusion Increased NLR in late stage of pregnancy could serve as an effective factor to predict the risk of preterm labor.

In physiological conditions, a diverse microbiota might enhance host defense. However, the gut microbiota of critically ill patients is characterized by lower diversity, lower abundances of key commensal genera, and overgrowth by one bacterial generation, a state known as dysbiosis. Increasing evidences indicate that microbiota-derived components can reach the systemic circulation from the gut and modulate immune homeostasis. Dysbiosis could have greater consequences for the critically ill patients and might contribute to poor outcome. In this review, we highlighted the crucial role of intestinal microbiota in systemic homeostasis in the critically ill patients and summarized emerging evidence in the field of microbiota-targeted therapies. This would provide new perspective for further establishing the causes and consequences of dysbiosis found in the critically ill patients as well as developing new strategies of intervention.