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.

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 assess the suitability of origin habitats of Notopterygium incisum, and provide theoretical basis of screening suitable areas for its large-scale cultivation.

METHODDetailed field survey of N. incisum, spatial databases, and GIS technology were used for the habitats suitability assessment.

RESULTMore than 142 073 km2 in 118 counties of Sichuan, Tibetan, Qinghai and Gansu are the most suitable habitats for N. incisum in which more than 47% of the area is located in sichuan, and more than of 377 000 km2 in 266 counties are relative suitable for N. incisum in Sichuan, Tibetan, Qinghai, Gansu, Yunnan, Xinjiang, etc and 32% of the area is located in Sichuan.

CONCLUSIONAlmost all the most suitable habitats are appropriate for germplasm conservation, wild population protection and regeneration due to the shortage of arable land, fragmentation of these alpine and subalpine ecosystems and sensitiveness of the environment changes. Therefore, large-scale cultivation of N. incisum could be developed in those relative suitable areas abundant in arable lands and labors, especially in moutainous regions with high elevation in the west of Sichuan province, and Qinghai Tibet plateau in the northwest of Sichuan, Southeast of Qinghai and Gansu province.

Shennongjia is well known as the national treasure of traditional Chinese medicine resources at home and abroad. In order to provide Shennongjia with better protection of traditional Chinese medicine and promote the sustainable utilization of its resources, based on the specific analysis of She nnongj ia′s conservation status and
available resources, we put forward primary strategies and specific measures for the sustainable utilization framework of Shennongjia traditional Chinese medicine resources, in view of resources conservation, development, utilization, as well as the resources administration.

Objective To observe the CALR mutation in patients with Ph negative chronic myeloproliferative neoplasms(MPNs) and its clinical significance. Methods From January 2012 to January 2015,the clinical data of ninety-seven patients with chronic myeloproliferative neoplasms was retrospectively analyzed and followed up to analyze different types of MPNs, including the clinical characteristics and gene mutation of polycythemia vera(PV),essential thrombocythemia(ET)and primary myelofibrosis (PMF).The hematological parameters and prognosis of patients with different mutation types were compared ( Cox regression model). Results Among the patients,the incidence of JAK2 mutation was the highest,64. 95% (63/97), followed by CALR mutation ( 19. 59% ( 19/97 ) ) and triple negative ( 10. 31% ( 10/97 ) ) . The incidence of MPL mutation was 5. 15% (5/97),which was the lowest and CALR mutations in ET and PMF were 28. 57%(10/35) and 28. 13% (9/32),respectively. The difference was not statistically significant (χ2 =1. 616,P>0. 05);the CALR gene mutation was not detected in PV patients. Compared with the JAK2 mutation, the hemoglobin,leukocyte and neutrophils in the patients with CALR mutation were lower (P<0. 05),PLT levels were lower in CALR-mutant ET patients ( P<0. 017) ,whereas platelet levels in CALR-mutant PMF patients were higher (P<0. 017). The incidence of disease progression in JAK2 and CALR mutation was 47. 62% (30/63)and 31. 58% (6/19) (χ2=1. 525,P>0. 05). The risk of disease progression in patients with CALR mutation was significantly lower than that of JAK2 mutation ( HR=0. 46,95%CI 0. 26-0. 98,P<0. 05) . Conclusion The clinical characteristics of MPNs patients with different gene mutations are different. The prognosis of MPNs patients with CALR mutation is better than that of JAK2 mutation.

Objective:To investigate the effect of different concentrations of human adipose-derived mesenchymal stromal cells (hADMSCs) derived exosome-mimetic nanovesicles (NVs)and exosomes (EXOs) on the biological function of human umbilical vein endothelial cells (HUVECs) .Methods:(1) Through hydrodynamic liposuction, adipose tissue was obtained from the thighs of 10 women (aged 18-65 years) in Fujian Medical University Union Hospital from June 2019 to August 2020. The hADMSCs were isolated by enzymatic hydrolysis, cultured to passage 4 and induced into adipocytes and osteocytes. The surface protein markers were identified by flow cytometry. (2) hADMSCs-NVs and hADMSCs-EXOs were prepared and observed under an electron microscope. Their surface protein markers were analyzed with particle size analyzer, particle size was analyzed with nanoparticle tracker. Protein quantitative analysis and nanoparticle tracking were used to detect the total protein and particle number of NVs and EXOs produced by 1×10 6 hADMSCs. (3) The control group (DMEM basic medium), 40, 60, 80 μg/ml NVs groups and 20, 40, 60 μg/ml EXOs groups were set to compare the proliferation, migration and angiogenesis of HUVECs through CCK-8 proliferation test, cell scratch test and angiogenesis test respectively. Graphpad Prism 7.0 was used for statistical analysis, and the measurement data were expressed as Mean±SD. Repeated measurement analysis of variance was applied to the comparison between multiple groups, and Tukey test was applied to pairwise comparison. P<0.05 represented statistical significance. Results:(1) The fourth generation of hADMSCs were slender spindle-shaped cells under optical microscope. After 21 days of adipogenesis induction, the transparent lipid droplets inside the cells were stained red by oil red O staining. After 14 days of osteogenesis induction, a large proportion of brown black staining area was observed by alkaline phosphatase calcium cobalt staining. The surface protein markers CD90 and CD29 of hADMSCs were positive. (2) Under transmission electron microscope, the structures of hADMSCs-NVs and EXOs were similar, both were discoid vesicles. The expression levels of CD9, CD81 and IgG were similar between NVS and EXOs. The particle sizes of NVs and EXOs were about the same, which were (72.0 ± 21.51) nm and (81.27±22.37) nm. The total protein content of NVs produced by 1×10 6 hADMSCs was (140.7±5.1) μg, about 100 times that of EXOs, which was (1.3±0.3) μg. The number of NVs [(644.5 ± 17.1)×10 8/ml] particles was about 90 times that of EXOs [(7.1±0.1)×10 8/ml]. (3) In CCK-8 proliferation assay, at 12, 24 and 48 hours after culture, the growth trend of HUVECs in the groups were generally consistent, and the difference in absorbance value was statistically significant ( P<0.01); at 48 hours after culture, the absorbance values of 60 μg/ml NVs and 40 μg/ml EXOs were higher than those in the control group (all P<0.01), but there was no significant difference between the two experimental groups ( P>0.05). At 8 and 24 hours after cell scratch assay, the changes of scratch width in each group were different, and the difference was statistically significant ( P<0.01); at 24 hours after scratch, the change of scratch width in 60 μg/ml NVs and 40 μg/ml EXOs groups were greater than that in the control group (all P<0.01), but there was no significant difference between the two experimental groups ( P>0.05). In the angiogenesis assay, the number of branch points and the length of blood vessels in each group were different, and the difference was statistically significant ( P<0.01). The number of capillary branches formed by HUVECs in 60 μg/ml NVs and 40 μg/ml EXOs groups were higher than that in the control group (all P<0.01), but there was no significant difference between the two experimental groups (all P>0.05). The capillary length of 60 μg/ml NVs and 40 μg/ml EXOs groups were longer than that of the control group ( all P<0.01), but there was no significant difference between the two experimental groups ( P>0.05). Conclusions:The shape and size of NVs were similar to EXOs, while the total protein content of NVs was about 100 times that of EXOs. The effects of hADMSCs-NVs and EXOs on the biological functions of HUVECs are similar and the optimum concentrations of NVs and EXOs are 60 μg/ml and 40 μg/ml, respectively.

Objective:To investigate the effect of different concentrations of human adipose-derived mesenchymal stromal cells (hADMSCs) derived exosome-mimetic nanovesicles (NVs)and exosomes (EXOs) on the biological function of human umbilical vein endothelial cells (HUVECs) .Methods:(1) Through hydrodynamic liposuction, adipose tissue was obtained from the thighs of 10 women (aged 18-65 years) in Fujian Medical University Union Hospital from June 2019 to August 2020. The hADMSCs were isolated by enzymatic hydrolysis, cultured to passage 4 and induced into adipocytes and osteocytes. The surface protein markers were identified by flow cytometry. (2) hADMSCs-NVs and hADMSCs-EXOs were prepared and observed under an electron microscope. Their surface protein markers were analyzed with particle size analyzer, particle size was analyzed with nanoparticle tracker. Protein quantitative analysis and nanoparticle tracking were used to detect the total protein and particle number of NVs and EXOs produced by 1×10 6 hADMSCs. (3) The control group (DMEM basic medium), 40, 60, 80 μg/ml NVs groups and 20, 40, 60 μg/ml EXOs groups were set to compare the proliferation, migration and angiogenesis of HUVECs through CCK-8 proliferation test, cell scratch test and angiogenesis test respectively. Graphpad Prism 7.0 was used for statistical analysis, and the measurement data were expressed as Mean±SD. Repeated measurement analysis of variance was applied to the comparison between multiple groups, and Tukey test was applied to pairwise comparison. P<0.05 represented statistical significance. Results:(1) The fourth generation of hADMSCs were slender spindle-shaped cells under optical microscope. After 21 days of adipogenesis induction, the transparent lipid droplets inside the cells were stained red by oil red O staining. After 14 days of osteogenesis induction, a large proportion of brown black staining area was observed by alkaline phosphatase calcium cobalt staining. The surface protein markers CD90 and CD29 of hADMSCs were positive. (2) Under transmission electron microscope, the structures of hADMSCs-NVs and EXOs were similar, both were discoid vesicles. The expression levels of CD9, CD81 and IgG were similar between NVS and EXOs. The particle sizes of NVs and EXOs were about the same, which were (72.0 ± 21.51) nm and (81.27±22.37) nm. The total protein content of NVs produced by 1×10 6 hADMSCs was (140.7±5.1) μg, about 100 times that of EXOs, which was (1.3±0.3) μg. The number of NVs [(644.5 ± 17.1)×10 8/ml] particles was about 90 times that of EXOs [(7.1±0.1)×10 8/ml]. (3) In CCK-8 proliferation assay, at 12, 24 and 48 hours after culture, the growth trend of HUVECs in the groups were generally consistent, and the difference in absorbance value was statistically significant ( P<0.01); at 48 hours after culture, the absorbance values of 60 μg/ml NVs and 40 μg/ml EXOs were higher than those in the control group (all P<0.01), but there was no significant difference between the two experimental groups ( P>0.05). At 8 and 24 hours after cell scratch assay, the changes of scratch width in each group were different, and the difference was statistically significant ( P<0.01); at 24 hours after scratch, the change of scratch width in 60 μg/ml NVs and 40 μg/ml EXOs groups were greater than that in the control group (all P<0.01), but there was no significant difference between the two experimental groups ( P>0.05). In the angiogenesis assay, the number of branch points and the length of blood vessels in each group were different, and the difference was statistically significant ( P<0.01). The number of capillary branches formed by HUVECs in 60 μg/ml NVs and 40 μg/ml EXOs groups were higher than that in the control group (all P<0.01), but there was no significant difference between the two experimental groups (all P>0.05). The capillary length of 60 μg/ml NVs and 40 μg/ml EXOs groups were longer than that of the control group ( all P<0.01), but there was no significant difference between the two experimental groups ( P>0.05). Conclusions:The shape and size of NVs were similar to EXOs, while the total protein content of NVs was about 100 times that of EXOs. The effects of hADMSCs-NVs and EXOs on the biological functions of HUVECs are similar and the optimum concentrations of NVs and EXOs are 60 μg/ml and 40 μg/ml, respectively.

The quality of Chinese herbal medicine is closely related to its producing region. In order to apply mathematical models to do a quantitative study on the suitability of Chinese herbal medicine, it is necessary to study on the ecological factors and the interpolation of climatic data, which influence the Chinese herbal medicine growth. The paper firstly studied the judgment standard of ecological index from the points of ecology and statistics, and how to calculate the optimum range values and the weight of each ecological factor. Secondly, meteorological element data is essential data in analyzing the suitable region of Chinese herbal medicine, and the spatial distribution of meteorological elements is closely related to terrain environment, so, in order to make the results close to true value by the greatest degree. The paper adopted multiple linear regression interpolation method which based on DEM. The paper distinguished the factor system of suitable region and interpolation on the point of datumization, and made a study on it about some key issues.
Adaptation, Biological ; Drugs, Chinese Herbal ; Ecology ; Environment ; Geographic Information Systems ; Models, Theoretical

Objective To investigate the prognostic factors of patients with upper urinary tract urothelial carcinoma (UTUC) treated with gemcitabine plus cisplatin (GC).Methods The clinical and follow-up data of 80 patients with UTUC admitted to Beijing Friendship Hospital,Capital Medical University from January 2013 to July 2018 were retrospectively analyzed.All patients underwent UTUC radical surgery.All patients were treated with GC regimen:1,8,and 15 days,Gemcitabine 800 mg/m2,intravenous infusion over 30 min;day 2 Cisplatin 70 mg/m2,protected from light 2 h intravenous drip;28 d for 1 cycle.Adjuvant treatments such as acid suppression,hydration,and antiemetic were given before and after chemotherapy.Patients completed 1 to 5 cycles with an average of 2 cycles.The patient's age,gender,presence or absence of water,primary tumor site,tumor stage and grade,lymphatic vascular infiltration,tumor recurrence,lymph node metastasis,organ metastasis,chemotherapy cycle,total Survival,etc.are used as indicators ofobservation.Univariate analysis of the patient's overall survival,screening for clinical variables associated with prognosis,and then using the COX proportional hazards model for multivariate prognostic analysis to determine independent influencing factors.Results Eighty patients with UTUC were followed up for 2 to 72 months with a median follow-up of 27 months.Sixteen patients (20％) died of UTUC recurrence or metastasis,and 64 (80％) patients survived.The 1-year cumulative survival rate was 78.26％ (18/23),and the 2-year cumulative survival rate was 54.18％ (9/13 ×78.26％),the 3-year cumulative survival rate was 39.41％ (8/1 1 × 54.18％),the 4-year cumulative survival rate was 31.53％ (12/15 × 39.41％),and the 5-year cumulative survival rate was 28.66％ (10/11 × 31.53％).Univariate analysis showed combined hydronephrosis (P =0.023),lymphatic vessel infiltration (LVI) (P =0.001),tumor TNM stage (P =0.002),tumor recurrence (P =0.008),simple lymph node metastasis (P =0.005),organ metastasis (P ＜ 0.001) was related to survival rate.COX model multivariate analysis showed that the independent risk factors associated with survival of patients with UTUC receiving chemotherapy with GC regimen were hydronephrosis (HR =4.355,95％CI:1.232-15.390,P=0.022),LVI (HR =0.133,95％ CI:0.035-0.509,P=0.003),TNM stage (HR=0.099,95％CI:0.010-0.929,P=0.043).Conclusion The presence or absence of hydronephrosis,LVI,and tumor TNM staging are independent factors influencing the prognosis of patients with UTUC who have adjuvant chemotherapy.