Integrated metabolome and transcriptome analyses reveal the molecular mechanism underlying dynamic metabolic processes during taproot development of Panax notoginseng.

BACKGROUND: Panax notoginseng (Burk) F. H. Chen is one of the most famous Chinese traditional medicinal plants. The taproot is the main organ producing triterpenoid saponins, and its development is directly linked to the quality and yield of the harvested P. notoginseng. However, the mechanisms underlying the dynamic metabolic changes occurring during taproot development of P. notoginseng are unknown. RESULTS: We carried out metabolomic and transcriptomic analyses to investigate metabolites and gene expression during the development of P. notoginseng taproots. The differentially accumulated metabolites included amino acids and derivatives, nucleotides and derivatives, and lipids in 1-year-old taproots, flavonoids and terpenoids in 2- and 3-year-old taproots, and phenolic acids in 3-year-old taproots. The differentially expressed genes (DEGs) are related to phenylpropanoid biosynthesis, metabolic pathway and biosynthesis of secondary metabolites at all three developmental stages. Integrative analysis revealed that the phenylpropanoid biosynthesis pathway was involved in not only the development of but also metabolic changes in P. notoginseng taproots. Moreover, significant accumulation of triterpenoid saponins in 2- and 3-year-old taproots was highly correlated with the up-regulated expression of cytochrome P450s and uridine diphosphate-dependent glycosyltransferases genes. Additionally, a gene encoding RNase-like major storage protein was identified to play a dual role in the development of P. notoginseng taproots and their triterpenoid saponins synthesis. CONCLUSIONS: These results elucidate the molecular mechanism underlying the accumulation of and change relationship between primary and secondary metabolites in P. notoginseng taproots, and provide a basis for the quality control and genetic improvement of P. notoginseng.

Diversity of fungal community and quality evaluation of Spatholobus Suberectus Dunn during the process of mildew.

Spatholobus suberectus Dunn as a traditional Chinese herbal medicine, which is susceptible to being infected by molds during storage. In order to explore the diversity characteristics of fungal community and the quality evaluation of Spatholobus suberectus Dunn during the process of mildew. The study used high-throughput sequencing technology to detect the diversity characteristics of fungal community, high-performance liquid chromatography (HPLC) and ultraviolet spectrophotometry (UV-spectrophotometry) methods to detect the content of flavonoids, and enzyme-linked immunosorbent assay (ELISA) method to detect the content of Aflatoxins B1 (AFB1). The result showed that the fungi of all samples belonged to 14 phyla, 336 genera, and the dominant fungi at the early stage of mildew was not obvious, while that at middle and late stages of mildew was Aspergillus. The species diversity of fungal community was the highest at the early stage of mildew, while the species richness of fungal community was the highest at the late stage of mildew. The content of AFB1 showed an upward trend, while the content of flavonoids showed a downward trend during the process of mildew. In brief, the diversity of fungal community decreased gradually, and the number of dominant fungi increased gradually, and the quality of Spatholobus suberectus Dunn decreased gradually during the process of mildew.

Characteristics of heavy metal accumulation and risk assessment in understory Panax notoginseng planting system.

Yunnan Province is the main planting area of the precious Chinese herbal medicines (CHM) Panax notoginseng; however, it locates the geological area with high soil heavy metals in China. The frequent land replacement due to continuous cropping obstacles and excessive application of chemicals makes P. notoginseng prone to be contaminated by heavy metals under the farmland P. notoginseng (FPn) planting. To overcome farmland shortage, understory P. notoginseng (UPn) was developed as a new ecological planting model featured by no chemicals input. However, this newly developed planting system requires urgently the soil-plant heavy metal characteristics and risk assessment. This study aimed to evaluate the pollution status of eight heavy metals in the tillage layer (0-20 cm), subsoil layer (20-40 cm) and the plants of UPn in Lancang County, Yunnan Province. Pollution index (Pi) showed that the contamination degree of heavy metals in the tillage layer and subsoil layer was Cd > Pb > Ni > Cu > Zn > Cr > Hg > As and Pb > Cd > Cu > Ni > Cr > Hg > Zn > As, respectively. Potential ecological risk index (PERI) for the tillage layer and subsoil layer was slight and middle, respectively. The exceeding standard rate of Cd, As, Pb, Hg, Cu in the UPn roots was 5.33%, 5.33%, 13.33%, 26.67% and 1.33%, respectively, while only Cd and Hg in the UPn leaves exceeded the standard 10% and 14%, respectively. The enrichment abilities of Cd and Hg in the roots and leaves of UPn were the strongest, while that of Pb was the weakest. The Hazard index (HI) and target hazard quotient (THQ) of eight heavy metals in the roots and leaves of UPn were less than 1.Therefore, our results prove that Upn has no human health risk and provide a scientific basis for the safety evaluation and extension of UPn.

Internal phosphorus cycling in macrophyte-dominated eutrophic lakes and its implications.

Macrophyte-dominated eutrophication (MDE) generally exhibits different characteristics from phytoplankton-dominated eutrophication (PDE). However, the significance of P cycling on eco-environmental management of MDE lakes is still not fully recognized. In this study, P-cycling mechanism was studied in a typical MDE lake (Wuliangsu Lake, China) based on a Before-After-Control-Impact design, taking advantage of the contrasting states between its 13 sub-lakes (with/without macrophytes and with/without external water flow). Our study demonstrated that P cycling predominantly occurs through "macrophyte ↔ sediment" in the MDE lakes, rather than "water ↔ sediment" as in PDE lakes; the biodynamics of the macrophytes acts as a primary driving force for this self-enforced P cycling. Our findings challenge the present lake eutrophication management strategies, which primarily limited to the water nutrient content, and demonstrate that successful MDE lake restoration should focus on stressors caused by the sustainable "macrophyte-sediment" P cycling. Macrophyte harvesting immediately before withering is recommended as the most sustainable technique in environment management for periodically frozen shallow MDE lakes. By this technique, sediment P can be gradually pumped up by the overgrown macrophytes each year until the advent of an alternative stable state (low sediment P, small biomass, and higher diversity), thereby forming a positive feedback loop "macrophyte harvesting → less sediment P → less macrophyte → higher diversity." Also, the catastrophic shift from MDE to PDE is no longer a concern. Furthermore, the macrophyte debris will not pose a problem as long as the macrophytes are removed during the harvest.

Ethanol Extract of Caesalpinia decapetala Inhibits Influenza Virus Infection In Vitro and In Vivo.

Influenza virus infections can lead to viral pneumonia and acute respiratory distress syndrome in severe cases, causing significant morbidity and mortality and posing a great threat to human health. Because of the diversity of influenza virus strains and drug resistance to the current direct antiviral agents, there have been no effective drugs as yet to cure all patients infected by influenza viruses. Natural products from plants contain compounds with diverse structures that have the potential to interact with multiple host and virus factors. In this study, we identified the ethanol extract of Caesalpinia decapetala (Roth) Alston (EEC) as an inhibitor against the replication of a panel of influenza A and B viruses both on human pulmonary epithelial A549 and human monocytic U937 cells. The animal study revealed that EEC administration reduces the weight loss and improves the survival rate of mice infected with lethal influenza virus. Also, EEC treatment attenuated lung injury and reduced virus titer significantly. In conclusion, we showed that EEC has antiviral activity both in vitro and in vivo, suggesting that the plant C. decapetala has the potential to be further developed as a resource of new anti-influenza drugs.

The Establishment and Validation of the Human U937 Cell Line as a Cellular Model to Screen Immunomodulatory Agents Regulating Cytokine Release Induced by Influenza Virus Infection.

Severe influenza infections are often associated with the excessive induction of pro-inflammatory cytokines, which is also referred to as "cytokine storms". Several studies have shown that cytokine storms are directly associated with influenza-induced fatal acute lung injury and acute respiratory distress syndrome. Due to the narrow administration window, current antiviral therapies are often inadequate. The efforts to use immunomodulatory agents alone or in combination with antiviral agents in the treatment of influenza in animal models have resulted in the achievement of protective effects accompanied with reduced cytokine production. Currently, there are no immunomodulatory drugs for influenza available for clinical use. Animal models, despite being ideal to study the anti-inflammatory responses to influenza virus infection, are very costly and time-consuming. Therefore, there is an urgent need to establish fast and economical screening methods using cell-based models to screen and develop novel immunomodulatory agents. In this study, we screened seven human cell lines and found that the human monocytic cell U937 supports the replication of different subtypes of influenza viruses as well as the production of the important pro-inflammatory cytokines and was selected to develop the cell-based model. The U937 cell model was validated by testing a panel of known antiviral and immunomodulatory agents and screening a drug library consisting of 1280 compounds comprised mostly of FDA-approved drugs. We demonstrated that the U937 cell model is robust and suitable for the high-throughput screening of immunomodulators and antivirals against influenza infection.

Nitrogen variations during the ice-on season in the eutrophic lakes.

Nitrogen accumulation in sediments, and the subsequent migration and transformations between sediment and the overlying water, plays an important role in the lake nitrogen cycle. However, knowledge of these processes are largely confined to ice-free seasons. Recent research under ice has mainly focused on the water eco-environmental effects during winter. Sediment N accumulation during the ice-on season and its associated eco-environmental impacts have never been systematically investigated. To address these knowledge gaps, we chose Wuliangsu Lake in China as a case study site, taking advantage of the spatial disparity between the 13 semi-separated sub-lakes. Based on samples of 35 sampling sites collected before, in the middle, and at the end of ice-on season separately, we performed a quantitative analysis of under-ice lake N accumulation and water-sediment N exchange by analyzing N fraction variations. Hierarchical Cluster Analysis and Relevance Analysis were used to help elucidate the main causes and implications of under-ice N variation. Our results clearly show that existing studies have underestimated the impact of under-ice N accumulation on the lake ecology throughout year: 1) Sediment N accumulated 2-3 times more than that before winter; 2) residual nitrogen (Res-N) contributed to the majority of the accumulated sediment N and was mainly induced by the debris of macrophytes; 3) total available nitrogen (TAN) was the most easily exchanged fractions between sediment and water, and it mainly affected the water environment during winter; 4) the Res-N accumulation during the ice-on season may have a strong impact on the eco-environment in the subsequent seasons. Our research is valuable for understanding the mechanism of internal nutrient cycle and controlling the internal nitrogen pollution, especially in shallow seasonally-frozen lakes that have long suffered from macrophyte-phytoplankton co-dominated eutrophication.

The relationship of lung cancer with menstrual and reproductive factors may be influenced by passive smoking, cooking oil fumes, and tea intake: A case-control study in Chinese women.

The aim of this study was to investigate the association of menstrual and reproductive factors with risk of lung cancer in women. Potential etiological clues related to lung cancer in women are identified to inform preventive strategies.Case-control study of 477 newly diagnosed women with lung cancer and 479 age-matched (±2 years) controls. Data on menstrual and reproductive factors and history of oral contraceptive use were obtained on personal interviews using a structured questionnaire. Risk factors were analyzed by unconditional logistic regression analysis.Maternal age ≥25 years at first birth appeared to protect against female lung cancer [odds ratios (ORs): 0.511, 95% confidence interval (CI), 0.376-0.693]. Age at menopause > 50 years and use of contraceptives was associated with an increased risk of lung cancer in women (OR: 1.471, 95% CI, 1.021-2.119 and OR: 1.844, 95% CI: 1.111-3.061, respectively). Age ≥13 years at menarche was associated with a decreased risk of lung adenocarcinoma (OR: 0.563, 95% CI, 0.317-0.997). There was significant heterogeneity in the levels of cooking oil fume (COF) exposure (Pheterogeneity = .015). Higher levels of exposure to passive smoking, COF, and lack of tea intake were associated with an increased risk of lung cancer.Menstrual and reproductive factors are considered to play a role in the development of lung cancer in women. Exposure to passive smoking, COF, and lack of tea intake appeared to significantly modify the relationship.

Identification of hydrolyzable tannins (punicalagin, punicalin and geraniin) as novel inhibitors of hepatitis B virus covalently closed circular DNA.

The development of new agents to target HBV cccDNA is urgently needed because of the limitations of current available drugs for treatment of hepatitis B. By using a cell-based assay in which the production of HBeAg is in a cccDNA-dependent manner, we screened a compound library derived from Chinese herbal remedies for inhibitors against HBV cccDNA. Three hydrolyzable tannins, specifically punicalagin, punicalin and geraniin, emerged as novel anti-HBV agents. These compounds significantly reduced the production of secreted HBeAg and cccDNA in a dose-dependent manner in our assay, without dramatic alteration of viral DNA replication. Furthermore, punicalagin did not affect precore/core promoter activity, pgRNA transcription, core protein expression, or HBsAg secretion. By employing the cell-based cccDNA accumulation and stability assay, we found that these tannins significantly inhibited the establishment of cccDNA and modestly facilitated the degradation of preexisting cccDNA. Collectively, our results suggest that hydrolyzable tannins inhibit HBV cccDNA production via a dual mechanism through preventing the formation of cccDNA and promoting cccDNA decay, although the latter effect is rather minor. These hydrolyzable tannins may serve as lead compounds for the development of new agents to cure HBV infection.

[Effect of electroacupuncture at Zhongwan(CV 12) on skin microcirculatory blood perfusion units along the conception vessel in yang-deficiency volunteers].

OBJECTIVE: To observe the effect of electroacupuncture(EA) at Zhongwan(CV 12) on the energy metabolism along the conception vessel(CV) in volunteers with yang-deficiency constitution,and to explore the relationship of electroacupuncture regulation and body constitution. METHODS: Eighteen volunteers with mild constitution and 18 volunteers with yang-deficiency constitution were collected out of 200 students of Fujian University of TCM by body constitution questionnaire. Skin microcirculatory blood perfusion units (MBPU) at Danzhong (CV 17), Xiawan(CV 10) and Qihai(CV 6) of CV were measured by a laser Doppler flowmetry in the normal condition and after EA stimulation at Zhongwan(CV 12) for 20 min. RESULTS: (1)Before treatment, (1)MBPU values at Danzhong(CV 17), Xiawan(CV 10) and Qihai(CV 6) in the yang-deficiency constitution group were lower than those in the mild constitution group,but there was no statistical significance (both P>0. 05) except Danzhong(CV 17) (P<0. 01). (Z)As for the three acupoints in the mild constitution group, MBPU level of Danzhong(CV 17) was higher than that of Xiawan(CV 10) without statistical significance(P->0. 05),and MBPU values of Danzhong(CV 17) and Xiawan(CV 10) were higher than that of Qihai(CV 6) (both P<0. 01). (3About the three acupoints in the yang-deficiency constitution group, MBPU result of Danzhong(CV 17) was lower than the value of Xiawan(CV 10), but higher compared with Qihai(CV 6)(P<0. 05, P<0. 01). MBPU of Xiawan(CV 10) was higher than Qihai (CV 6) as well(P<0. 01). (2) MBPU values of Danzhong(CV 17), Xiawan(CV 10) and Qihai(CV 6) were increased apparently compared with those before treatment after EA stimulation at Zhongwan(CV 12) for 20 min in the two groups(all P<0. 01). (3) The rise rates of MBPU level about Danzhong(CV 17) and Qihai(CV 6) in the yang-deficiency constitution group were higher than those in the mild constitution group without statistical significance after EA at Zhongwan(CV 12) for 20 min(both P>0. 05). CONCLUSION: The energy metabolism in CV of volunteers with yang-deficiency constitution is declined, especially Danzhong(CV 17). EA can rise energy metabolism in CV of mild or yang-deficiency constitution volunteers through regulating MBPU along meridian.

Application of parecoxib in post-uvulopalatopharyngoplasty analgesia.

OBJECTIVE: To investigate the postoperative analgesic effects of parecoxib for uvulopalatopharyngoplasty (UPPP). METHODS: Patients with obstructive sleep apnoea syndrome who underwent UPPP were randomly divided into two groups. In group A, the incision-local block was performed with 5 ml of 0.5% ropivacaine injected subcutaneously before the end of surgery, then 20 ml of physiological saline was injected intravenously every 12 h for 2 days. In group B, in addition to the incision-local block, 40 mg parecoxib was injected intravenously 30 min before the end of UPPP and 40 mg parecoxib was injected intravenously every 12 h for 2 days. Postoperative pain was measured using a visual analogue scale (VAS). Adverse reactions were recorded. RESULTS: A total of 40 patients were randomized (n = 20 per group). Under resting conditions, the mean ± SD VAS pain scores were significantly higher in group A compared with group B at 24 h and 48 h after UPPP (24 h 4.0 ± 0.8 versus 2.6 ± 0.6; 48 h 3.8 ± 0.7 versus 2.4 ± 0.5; respectively). Under swallowing conditions, the mean ± SD VAS pain scores were significantly higher in group A compared with group B at 8 h, 24 h and 48 h after UPPP. Postoperative adverse reactions were similar in the two groups. CONCLUSION: Intravenous parecoxib combined with incision-local ropivacaine provided effective postoperative analgesia for patients with obstructive sleep apnoea syndrome, undergoing UPPP.

[Isolation, purification and composition analysis of water-soluble polysaccharide from Halenia elliptica].

OBJECTIVE: To provide scientific cue for the use of Halenia elliptica. METHODS: The crude polysaccharide was extracted from Halenia elliptica with hot water and precipitated by ethanol. The crude polysaccharide has been eliminated protein and fractionated by acidic ethanol. Four fractions HM1, HM2, HM3 and HM4 were got respectively. RESULTS: By using DEAE-Sephadex A-25 gel filtration, HM1 was got. GC analysis indicated that HM1, HM2, HM3, HM4 and HM41 were composed of Rha, Ara, Xyl, Man, Gal and Glc. But their molar ratios were different. CONCLUSION: Polysaccharide HM41 is isolated from Halenia elliptica for the first time.

[Study on the purification and scavenging free radical activity of water soluble polysaccharide of leave in Hippohae rhamnoides L].

The crude polysaccharide was extracted from Leave in Hippohae rhamnoides L. with hot water, and precipitated by ethanol. The crude polysaccharide has been fractionated by acidic ethanol. Three fractions (SJ1, SJ2, SJ3) were got respectively. SJ2 deproteinizationed by the combined methods of enzyme and Seveage, purified by DEAE-Sephadex A-25 gel filtration. PC analysis indicated that SJ22 is composed of Xyl,Ara,Glc,Gal,GaL A. The identification of purify by Sepharose CL-4B, paper chromatography and cellulous acetate electrophoresis showed it was homogeneous. Typical absorption of polysaccharides was shown in its IR spectrum. It contained a-glucosidic bonds by IR analysis. It had typical absorption of protein by UV scaning. SJ22 is first isolated from Leave in Hippohae rhamnoides L Scavenging free radical experiment showed that SJ22 was effective in scavenging superoxideradical and hydroxylradical, but only a little effective in scavenging lipid radical.