Multi-omics-based identification of purple acid phosphatases and metabolites involved in phosphorus recycling in stylo root exudates.

Stylo (Stylosanthes guianensis) is a tropical forage and cover crop that possesses low phosphate (Pi) tolerance traits. However, the mechanisms underlying its tolerance to low-Pi stress, particularly the role of root exudates, remain unclear. This study employed an integrated approach using physiological, biochemical, multi-omics, and gene function analyses to investigate the role of stylo root exudates in response to low-Pi stress. Widely targeted metabolomic analysis revealed that eight organic acids and one amino acid (L-cysteine) were significantly increased in the root exudates of Pi-deficient seedlings, among which tartaric acid and L-cysteine had strong abilities to dissolve insoluble-P. Furthermore, flavonoid-targeted metabolomic analysis identified 18 flavonoids that were significantly increased in root exudates under low-Pi conditions, mainly belonging to the isoflavonoid and flavanone subclasses. Additionally, transcriptomic analysis revealed that 15 genes encoding purple acid phosphatases (PAPs) had upregulated expression in roots under low-Pi conditions. Among them, SgPAP10 was characterized as a root-secreted phosphatase, and overexpression of SgPAP10 enhanced organic-P utilization by transgenic Arabidopsis. Overall, these findings provide detailed information regarding the importance of stylo root exudates in adaptation to low-Pi stress, highlighting the plant's ability to release Pi from organic-P and insoluble-P sources through root-secreted organic acids, amino acids, flavonoids, and PAPs.

Evaluating influence of filling fraction of carriers packed in anaerobic side-stream reactors on membrane fouling and microbial community of the coupled membrane bioreactors.

An anoxic/oxic membrane bioreactors (AO-MBR) and three identical anaerobic side-stream reactor coupled with anoxic/oxic membrane bioreactors (ASSR-MBR) were constructed and operated in parallel to investigate the appropriate filling fraction of carriers packed in ASSR, influences on pollutants removal, sludge reduction, membrane fouling and microbial community of ASSR-MBR. Inserting ASSR achieved efficient COD removal and nitrification, and packing carriers in ASSR obtained the highest sludge reduction efficiency of 50.5 % at filling fraction of 25 %. Compared to AO-MBR, inserting ASSR without carriers induced the release of viscous components in extracellular polymeric substances (EPS) and the formation of calcium carbonate-related bacteria on membrane surface, and thus deteriorated membrane fouling. Packing carriers with 25 % filling fraction promoted the hydrolysis of soluble microbial products and EPS, whilst reduced the viscoelasticity of sludge flocs. Higher filling fraction of 50 % increased the shear forces to the biofilm and biomarkers related to membrane fouling, and thus showed little improvement to alleviate membrane fouling. MiSeq sequencing revealed that although it enriched in the bulk sludge of conventional ASSR-MBR and the coupled reactor with filling fraction of 50 %, the floc-forming, hydrolytic and fermentative bacteria were more inclined to attach on the membrane surface and alleviate fouling process.

[Optimization for phosphorous removal in thickening and dewatering sludge water by polyaluminum chloride].

Based on the comparison of phosphorous removal in sludge water and its supernatant by polyaluminum chloride (PAC), separate and combined effects of Al/P mole ratio, pH and mixing speed (MS) on phosphorus removal by PAC for the supernatant of thickening and dewatering sludge water were analyzed by the response surface methodology (RSM), and kinetics of phosphorous removal by PAC was also investigated. The results showed that direct addition of PAC into sludge water deteriorated its settling characteristics, and suspended solids in the sludge water could decrease the phosphorus removal efficiency. The RSM analysis results demonstrated that the effect of individual operation parameter on phosphorus removal was followed as the order of Al/P > pH > MS, and the optimal process parameters with phosphorus removal efficiency of 97.8% were Al/P = 2.49, pH = 8.3 and MS 398 r x min(-1), respectively. The verification experiment showed that the RSM model was valid and effective. Kinetic analysis illustrated that phosphorus removal by PAC was divided into two stages, a chemical precipitation and rapid adsorption stage, followed by a chemical precipitation stage that conformed to the second-order kinetics.

Lappaol F, a novel anticancer agent isolated from plant arctium Lappa L.

In an effort to search for new cancer-fighting therapeutics, we identified a novel anticancer constituent, Lappaol F, from plant Arctium Lappa L. Lappaol F suppressed cancer cell growth in a time- and dose-dependent manner in human cancer cell lines of various tissue types. We found that Lappaol F induced G(1) and G(2) cell-cycle arrest, which was associated with strong induction of p21 and p27 and reduction of cyclin B1 and cyclin-dependent kinase 1 (CDK1). Depletion of p21 via genetic knockout or short hairpin RNA (shRNA) approaches significantly abrogated Lappaol F-mediated G(2) arrest and CDK1 and cyclin B1 suppression. These results suggest that p21 seems to play a crucial role in Lappaol F-mediated regulation of CDK1 and cyclin B1 and G(2) arrest. Lappaol F-mediated p21 induction was found to occur at the mRNA level and involved p21 promoter activation. Lappaol F was also found to induce cell death in several cancer cell lines and to activate caspases. In contrast with its strong growth inhibitory effects on tumor cells, Lappaol F had minimal cytotoxic effects on nontumorigenic epithelial cells tested. Importantly, our data also demonstrate that Lappaol F exhibited strong growth inhibition of xenograft tumors in nude mice. Lappaol F was well tolerated in treated animals without significant toxicity. Taken together, our results, for the first time, demonstrate that Lappaol F exhibits antitumor activity in vitro and in vivo and has strong potential to be developed as an anticancer therapeutic.

Simultaneous determination of eight active components in Houttuynia cordata injection and its quality control in productive process.

A simple, reliable and effective gas chromatography coupled with flame ionization detection method was developed for the simultaneous determination of eight components (α-pinene, ß-pinene, myrcene, limonene, terpinen-4-ol, α-terpineol, bornyl acetate and methyl-n-nonylketone) in Chinese medicine Houttuynia cordata and its injection. The chromatographic separation of all eight components, including undecylene as internal standard was performed on a DB-1 column (30 m×0.25 mm, 0.25 µm). Excellent linear behaviors including herb and injection over the investigated concentration ranges were observed with the values of r(2) higher than 0.9990 for all analytes. Satisfactory intra-day and inter-day precisions were achieved with RSD less than 2% and the average recoveries for all analytes at three different concentrations obtained were in the range of 93.4-104.4%, with RSD ranging from 1.3 to 4.1%. The proposed method was successfully applied in the simultaneous determination of these active components in H. cordata and H. cordata injection (HCI), including the intermediate product of HCI in productive process, from different pharmaceutical factories and different production batches, indicating that the method in this paper was particularly suitable for the routine analysis of HCI and its quality control in productive process.

Treatment of polluted landscape lake water and community analysis of ammonia-oxidizing bacteria in constructed wetland.

Three lab-scale horizontal subsurface flow constructed wetlands (HSSF CWs) were installed to demonstrate the use of constructed wetlands as a viable low-cost treatment option to purify the polluted scenery water. The goals of this study were to determine the effectiveness of the wetland treatment technology in reducing contaminants in polluted natural water body and to characterize bacterial composition present in 3 constructed wetlands. Water samples were collected periodically for 12 months from three wetlands to determine the efficiency of the treatment system in removal of chemical pollutants. The reduction by the treatment was greatest for chemical oxygen demand (COD), total nitrogen and total phosphorus, but the removal of nutrients obviously varied with seasons. Plant uptake was highly responsible for nitrogen and phosphorus removal. Denaturing gradient gel electrophoresis (DGGE) was used to study the diversity of microbial community in the constructed wetland. Changes in the total bacterial community and ammonia-oxidizing bacterial composition were examined by DGGE and sequencing of polymerase chain reaction (PCR)-amplified fragments of the gene carrying the ammonia monooxygenase subunit A gene (amoA) recovered from matrix samples and DGGE bands. DGGE analysis of wetlands samples revealed that seasonal change had an effect on the diversity and composition of microbial communities in constructed wetlands. The sequence analysis showed ammonia-oxidizing bacteria (AOB) in constructed wetlands were uncultivable and the population of AOB had a higher percentage of Nitrosomonas-like sequences from wetlands, while no Nitrosospira-like sequences were found.

[Identification of Gynostemma from their adulterant Cayratia japonica by PCR-RFLP].

OBJECTIVE: To establish a convenient and effective method for the identification of Gynostemma and Cayratia japonica. METHOD: Eight species, including Gynostemm pentaphyllum, G. pentagynum, G. cardiospermum, G. longipe, G. yixingense, G. laxiflorum, G. guangxiense and C. japonica were investigated through PCR - RFLP of six chloroplast DNA fragments. The six gene fragments were digested by six restriction endonuclease respectively, including Taq I, Hpa II, EcoR I, Rsa I, Hha I, Hind III. RESULT: Seven species of Gynostemma and their adulterant could be identified by trnK1f-trnK2r and Rsa. CONCLUSION: PCR - RFLP provides a quick, reliable molecular marker technique for identification of Cynostemma and their adulterant Cayratia japonica.

[Microlaryngoscopic surgery in the treatment of vocal cord benign lesion under surface anesthesia].

OBJECTIVE: To evaluate the effect of microlaryngoscopic surgery in the treatment of vocal cord benign lesion. METHOD: The operation was performed using suspension laryngoscope and common operative microscope under surface anesthesia with 1% pontocaine. RESULT: In the 872 patients,438 cases had vocal nodules, 353 were vocal polyps and other vocal cord cysts. In 208 patients underwent successful operation, 171 cases were cured clinically and 36 cases improved after the first attempt with a general effective rate of 96.3%. CONCLUSION: The most patients who had vocal cord benign lesions without severe cardiovascular diseases could be operated under surface anesthesia which have the advantages of rare complications and rapid recovery.