This paper is aimed to establish the method of fingerprint analysis of chemical constituents by reversed-phase ultra-performance liquid chromatography (UPLC) for the quality control of the roots and rhizomes of Panax ginseng (Ginseng Radix et Rhizoma). The method was performed on a ACQUITY UPLC BEH C18 (50 mm x 2.1 mm ID, 1.7 microm) with a mixed mobile phase of water and acetonitrile in a gradient mode. The flow rate was 0.3 mL x min(-1) and the wavelength of measurement was 203 nm. Eleven batches of the Ginseng Radix et Rhizoma were determined. The UPLC chromatographic fingerprints of chemical constituents were established from the eleven batches of the Ginseng Radix et Rhizoma and showed fifteen characteristic common peaks, among which fifteen peaks were recognized and nine compounds (ginsenosides Rg1, Re, Rf, Rg2, Rb1, Rc, Rb2, Rb3 and Rd) were determined by comparison with chromatographic behaviors and UV spectra of the authentic compounds. The eleven batches of samples were classified as two clusters by hierarchical clustering analysis (HCA) and principle component analysis (PCA), and six samples were confirmed to establish the mutual model. The quality was assessed by similarity evaluation system for chromatographic fingerprint of TCM (2004B Version). The convenient and high specific method can be used to identify and evaluate the quality of the Ginseng Radix et Rhizoma.

RNA-Sequencing (RNA-Seq) is a newly-developed method in transcriptome research, it can afford more accurate transcription information and be more quickly by using Next-generation Sequencing (NGS) technology. RNA-Seq has been widely used in various biological fields. Genuine traditional Chinese medicines (TCM), with good quality and therapeutic effect, were always praised highly and used by famous physicians. The geo-herbalism formation of TCM is based on the product of the gene expression at specific space and time. So it has been a research hotspot to analyze the mechanism of biosynthesis through RNA-Seq in the study on the secondary metabolism of medicinal plant. This article mainly illustrates the RNA-Seq and its advantages, it also discusses the potential application in genuine TCM, and it can provide useful information for other researchers.

Objective To discuss the correlation between female chronic pelvic pain and pelvic floor anatomy.Methods The female patiems of chronic pelvic pain 179 cases,age 28-67 years,average 49.4 years;pelvic pain history 8 months-9 years,average 2.8 years;167 cases has childbirth history,43 cases has surgery history,which gynecological surgery 31 cases,and urinary surgery 7 cases,and anus surgery 5 cases.Results High incidence of female pelvic pain were 30-60 age (incidence of 54.8％),93.3％ has birth history,24％ has operation history,the myofascial tissue pain higher than the organ,were 87.4％ than 12.6％ (P ＜ 0.01),the front of pelvic pain higher than back,were 65.6％ than 21.8％ (P ＜ 0.01).Conclusions Female chronic pelvic pain associated with the particularity of the pelvic anatomy and physiological,with the tissue of pelvic floor and urogenital diaphragm of primary injure and chronic inflammation is an important cause of chronic pelvic pain.

Quality variation and ecotype classification of Chinese herbal medicine are important scientific problems in Daodi herbal medicine research. The diversity of natural environmental conditions has led to form unique multi-Daodi, multi-product areas that produce particular Chinese herbal medicine. China is one of three big American ginseng (Panax quinquefolium L.) producing areas worldwide, with over 300 years of application and 40 years of cultivation history. Long-term production practice has led to the formation of three big advocate produce areas in China: Northeast province, Beijing and Shandong. P. quinquefolium L. grown under certain environmental conditions will develop long-term adaptations that will lead to more stable strains (different ecotypes). P. quinquefolium L., can vary greatly in quality; however, the ecological mechanisms causing this variation are still unclear. Root samples were collected from four-year-old cultivated P. quinquefolium L. plants in the three major genuine (Daodi) American ginseng-producing areas of Northeast province, Beijing and Shandong province, China. Ultra-performance liquid chromatography was used to analyze the contents of eight ginsenosides (Rg1, Re, Rb1, Rb2, Rb3, Rc, Rd, Rg2). Data for nine ecological factors, including temperature, moisture and sunlight, were obtained from the ecological database of Geographic Information System for Traditional Chinese Medicine. Soil samples from the sampling sites were collected. Effective boron and iron, available nitrogen and potassium, as well as other trace elements and soil nutrients, were determined by conventional soil physicochemical property assay methods. Analytical methods of biostatistics and numerical taxonomy were used to divide ecotypes of the three main Panax quinquefolium L. producing areas in China based on ginsenoside content, climate, soil and other ecological factors. To our knowledge, this is the first time that ecological division of P. quinquefolium L. producing areas in China has ever been conducted. The results show that there are two chemoecotypes of P. quinquefolium L. in China: ginsenoside Rb1-Re from outside Shanhaiguan, and ginsenoside Rg2-Rd from inside Shanhaiguan. Similarly, there are two types of climatic characteristics: inside Shanhaiguan (Beijing, Shandong) and outside Shanhaiguan (Northeast). This suggests that the formation and differentiation of chemoecotypes of P. quinquefolium L. is closely related to variability of the climatic and geographical environment. Additionally, ecological variation of the three main producing areas, characteristics of two climatic ecotypes, and soil characteristics are also discussed and summarized. These results provide experimental scientific evidence of the quality variation and ecological adaptation of P. quinquefolium L. from different producing areas. They also deepen our understanding of the biological nature of Daodi P. quinquefolium L. formation, and offer novel research models for other multi-origin, multi-Daodi Chinese herbal medicines ecotypes. In addition, the results demonstrate the critical need for improving quality, appropriate ecological regionalization and promoting industrialized development of P. quinquefolium L.

ObjectiveTo investigate the clinical features of the abnormal chromosome karyotypes in twin pregnancies complicated with fetal malformations. Methods Totally 181 twin pregnancies (362 fetuses) in which one or two fetuses had abnormalities diagnosed by ultrasound were referred to the First Affiliated Hospital of Sun Yat-Sen University from January, 2000 to September, 2010. They were divided into different groups according to ( 1 ) maternal age: the cases with maternal age ≥35 were divided into advanced pregnancy group ( 105 fetuses) , and those with maternal age ＜35 were divided into young pregnancy group (203 fetuses) ; ( 2 ) conceived method : those conceived by assisted reproductive technology were divided into assisted reproductive group (81 fetuses), and the natural conception pregnancies were divided into natural conception group (227 fetuses) ; ( 3 ) chorionicity: the monochorionic twin (MCT) pregnancies were divided into MCT group( 123 fetuses), and the dichorionic twin (DCT) pregnancies were divided into DCT group( 185 fetuses); (4) structural abnormalities: 205 fetuses with structural abnormalities were divided into the abnormal fetal group, and 103 fetuses without structural abnormalities were divided into the normal fetal group. All fetuses were examined by the ultrasound and chromosomes were examined in 308 fetuses. Results( 1 ) The karyotype of fetuses: among 181 twin pregnancies, 23 cases had chromosomal abnormalities in 1 or 2 fetuses ( 12. 7％ ,23/181 ), and chromosomes were exarmined in both fetuses in 20 of 23 cases. Twenty-six of 308 fetuses were found with abnormal chromosomes ( 8.4％, 26/308 ) , and the aneuploid was the most common type of abnormal karyotypes ( 53.8％ , 14/26 ). Twenty-one of 205 fetuses with malformations were found with abnormal karyotypes (10. 2％, 21/205 ). (2) Seven of 123 fetuses in MCT group were with abnormal karyotypes (5.7％, 7/123), and 19 of 185 fetuses in DCT group were with abnormal karyotypes ( 10. 3％, 19/185 ). There was no statistical difference of abnormal chromosome incidence between the two groups. There were 14 fetuses with aneuploid in DCT group ( 7.6％, 14/185 ) ;but there was no fetus with aneuploid in MCT group. There was statistical difference between these two groups. In two cases of DCT group, only one fetus with malformation received chromosome examination because another fetus was dead, and the karyotypes were trisomy 21 and trisomy 18 respectively. Both fetuses of the rest 17 cases received chromosome examination, and the chromosomes of both fetuses in each pregnancy were different. Fifteen of 19 fetuses with abnormal chromosomes in DCT group were complicated with structural abnormalities, and 7 fetuses of 4 twin pregnancies in MCT group were with chromosomal abnormalities. (3) The comparison of the abnormal karyotype incidence between the advanced pregnancy group and young pregnancy group: the abnormal karyotype incidence of the advanced pregnancy group was 7. 6％ (8/105), and that was 8.9％ (18/203) in young pregnancy group. There was no statistical difference between the two groups ( P ＞ 0. 05 ). Six of 105 fetuses in advanced pregnancy group were aneuploids (5. 7％, 6/105), and 8 of 203 fetuses in young pregnancy group were aneuploids (3.9％, 8/203). The aneuploid incidence in advanced pregnancy group was significantly higher than that in young pregnancy group ( P ＜ 0. 05 ). (4) The comparison of the abnormal karyotype incidence between the assisted reproductive group and the natural conception group: 11 of 81 fetuses were with the abnormal karyotypes in assisted reproductive group ( 13. 6％, 11/81 ), and 15 of 227 fetuses were with the abnormal karyotypes in assisted reproductive group (6. 6％, 15/227). There was statistical difference between the two groups ( P ＜0. 05). There were 7 fetuses with the aneuploid in assisted reproductive group ( 8. 6％, 7/81 ) and 7 fetuses with the aneuploid in natural conception group ( 3. 1％, 7/227 ), which showed no statistical difference ( P ＞0. 05 ).(5) The comparison of the abnormal karyotype incidence between the abnormal fetal group and normal fetal group: 21 of 205 fetuses in abnormal fetal group were with abnormal karyotypes (10. 2％, 21/205), and 5 of 103 fetuses in normal fetal group were with abnormal karyotypes ( 4. 9％, 5/103 ) . There was no statistical difference (P ＞ 0. 05 ). 13 fetuses in abnormal fetal group were with the aneuploid (6. 3％, 1 3/205), and only one fetus in normal fetal group was aneuploid (1.0％, 1/103 ). There was statistical difference between the two groups ( P ＜ 0. 05 ). ConclusionsAneuploid is the most common abnormal karyotype in twin pregnancy complicated with fetal abnormalities, especially trisomy 21. Aneuploid mainly occurs in only one fetus of DCT, and chromosomal discordance is usually found in DCT. While in MCT, the twin fetuses with the same abnormal karyotype may have different pbenotypes. The results suggest that it is necessary to analyze both karyotypes of twins even if only one fetus is complicated with structural abnormalities.

To explore the analysis methods for composite traditional Chinese medicine (TCM) constitutions.

Objective To explore the genetic mechanism of Yang-deficiency constitution by detecting genomic copy number variations (CNVs). Methods Thirty cases of Yang-deficiency constitution and 30 cases of balanced constitution were included according to the standards of Classification and Determination of Constitution in Traditional Chinese Medicine. DNA was extracted from white blood cells in peripheral blood. A genome-wide association study was conducted by using Affymetrix SNP 6.0 platform. CNVs of each sample were analyzed using PennyCNV software. The Yang-deficiency constitution-specific copy number variation regions (CNVRs) of each autosome were identified. CNVR-related genes and their annotations were searched at online Human Genome Browser. Results The mean number of CNVs in balanced constitution group was 12.63±3.39, ranging from 8 to 20. After stepwise elimination of two Yang-deficiency constitution subjects, the mean number of CNVs in Yang-deficiency constitution group was 15.04±8.95, ranging from 2 to 38. A total of 26 CNVRs were identified from 28 Yang-deficiency constitution subjects, including 19 duplicated CNVRs, 6 deleted CNVRs, and 1 mixed type CNVR. Most CNVRs were shared by a few Yang-deficiency constitution subjects, and only 7 CNVRs were shared by more than 5 Yang-deficiency constitution subjects. The functions of representative genes in Yang-deficiency constitution-specific CNVRs were related with extracellular and intracellular signal transduction, metabolic regulation, and immune response, etc. Conclusion Yang-deficiency constitution subjects have some specific genomic CNVs, which might result in Yang-deficiency constitution phenotypes by influencing the expression of genes associated with extracellular and intracellular signal transduction, material metabolism (energy metabolism), and immune response, etc.

Objective To investigate the chemical constituents in the roots of Boehmeria nivea.Methods The constituents were isolated by repeated column chromatography and their structures were elucidated by chemical properties and spectroscopic analyses.Results Seven compounds were isolated and their structures were identified to be daucosterol-10,13-eicosdienoate(Ⅰ),daucosterol(Ⅱ),?-sitosterol(Ⅲ),olein(Ⅳ),betulinic acid(Ⅴ),oleanolic acid(Ⅵ),19?-hydroxyursolic acid(Ⅶ).Conclusion Compound Ⅰ is a new compound named niveain A,compound Ⅳ is obtained from the plants of Boehmeria Jacq.for the first time.

Objective To explore the effect of Exosomes isolated from the A549 lung cancer cells on the proliferation of these cells and their ho?mologous tumor cells,HCC827,and the role of the PI3K/Akt and SRC signaling pathways in this process. Methods Exosomes were isolated from the supernatant after density gradient centrifugation of A549 cells. The Exosomes morphology was observed by transmission electron microscopy. The expression of the Exosome?specific proteins was analyzed using Western blotting. Cell proliferation was investigated using the MTT assay. Re?sults The A549?derived Exosomes were 30?100 nm in diameter and had a bilayer membrane.Western blotting showed that CD9 was detected in these Exosomes. The isolated Exosomes promoted the proliferation of the A549 and the HCC827 cells in a dose?and time?dependent manner,ac?companied by the activation of Akt and SRC. Conclusion Exosomes isolated from A549 cells promote the proliferation of the secreting cells and the homologous tumor cells in a dose?and time?dependent manner. The mechanism may be related to the activation of Akt and SRC.

Objective To investigate the effect of Exosomes derived from lung cancer cells on the migration of secretory cells and homologous tumor cells and to explore the role of PI3K/Akt and SRC signaling pathways in this process.Methods Exosomes were isolated from the supematant post density gradient centrifugation of A549,lung cancer cells.Morphology of the Exosomes was studied using transmission electron microscopy.Protein expression was analyzed using Western blotting.Cell migration was analyzed by a transwell assay.Results The double-membrane-bound Exosomes appeared as discal-shaped structures,30-100 nm in diameter.Western blotting showed that CD9 was abundant in the Exosomes.The Exosomes promoted the migration of A549 cells and their homologous tumor cells,HCC827 in a dose-dependent manner,accompanied by the activation of Akt and SRC.Conclusion The Exosomes derived from A549 (lung cancer) cells promote the migration of the secreting cells and the homologous tumor cells.The mechanism may be correlated with the activation of Akt and SRC.