Genomic and transcriptomic studies on flavonoid biosynthesis in Lagerstroemia indica.

BACKGROUND: Lagerstroemia indica is a widely cultivated ornamental woody shrub/tree of the family Lythraceae that is used as a traditional medicinal plant in East Asia and Egypt. However, unlike other ornamental woody plants, its genome is not well-investigated, which hindered the discovery of the key genes that regulate important traits and the synthesis of bioactive compounds. RESULTS: In this study, the genomic sequences of L. indica were determined using several next-generation sequencing technologies. Altogether, 324.01 Mb sequences were assembled and 98.21% (318.21 Mb) of them were placed in 24 pseudo-chromosomes. The heterozygosity, repeated sequences, and GC residues occupied 1.65%, 29.17%, and 38.64% of the genome, respectively. In addition, 28,811 protein-coding gene models, 327 miRNAs, 552 tRNAs, 214 rRNAs, and 607 snRNAs were identified. The intra- and interspecies synteny and Ks analysis revealed that L. indica exhibits a hexaploidy. The co-expression profiles of the genes involved in the phenylpropanoid (PA) and flavonoid/anthocyanin (ABGs) pathways with the R2R3 MYB genes (137 members) showed that ten R2R3 MYB genes positively regulate flavonoid/anthocyanin biosynthesis. The colors of flowers with white, purple (PB), and deep purplish pink (DPB) petals were found to be determined by the levels of delphinidin-based (Dp) derivatives. However, the substrate specificities of LiDFR and LiOMT probably resulted in the different compositions of flavonoid/anthocyanin. In L. indica, two LiTTG1s (LiTTG1-1 and LiTTG1-2) were found to be the homologs of AtTTG1 (WD40). LiTTG1-1 was found to repress anthocyanin biosynthesis using the tobacco transient transfection assay. CONCLUSIONS: This study showed that the ancestor L. indica experienced genome triplication approximately 38.5 million years ago and that LiTTG1-1 represses anthocyanin biosynthesis. Furthermore, several genes such as LiDFR, LiOMTs, and R2R3 LiMYBs are related to anthocyanin biosynthesis. Further studies are required to clarify the mechanisms and alleles responsible for flower color development.

A novel type lavandulyl flavonoid from Sophora flavescens as potential anti-hepatic injury agent that inhibit TLR2/NF-κB signaling pathway.

ETHNOPHARMACOLOGY RELEVANCE: Sophora flavescens Aiton, was a crucial source of Traditional Chinese Medicine (TCM) that has benefited human health for hundreds of years. Alkaloids and flavonoids were the major bioactive constituents from S. flavescens, which had been widely used for liver disease treatment in China. However, the liver-protective components of flavonoids from S. flavescens and their mechanism of action were not clear. AIM OF THE STUDY: This work aimed to evaluate the in vitro hepatoprotective activities of 35 flavonoids from S. flavescens and screen active compounds. Furthermore, it was conducted to demonstrate the hepatoprotective effects of a new active compound (kurarinol A, 1) was isolated by authors and the ethyl acetate (EtOAc) extract form S. flavescens against carbon tetrachloride (CCl4)-induced hepatic injury in Kunming (KM) mice, meanwhile revealed the potential mechanism. MATERIALS AND METHODS: The 35 flavonoids from S. flavescens were co-incubated with HepG2 cells and treated with 0.35% CCl4 for 6 h cell viability was measured by (3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium, inner salt) (MTS) assay. Then, in vivo animal experiments, the activities of alanine aminotransferase (ALT), aspartate aminotransferase (AST) and alkaline phosphatase (ALP) in the serum were analyzed, the degree of hepatic injury was examined using hematoxylin-eosin (H&E) staining, the mRNA expression of Superoxide Dismutase 2 (SOD2), Nuclear factor E2-related factor 2 (Nrf2), heme oxygenase 1 (HO-1), Interleukin 6 (IL-6), Tumor Necrosis Factor-α (TNF-α), interleukin-1ß (IL-1ß), and the protein levels of nuclear factor-kappa B p65/p-p65 (NF-κB p65/p-p65), toll-like receptor 2 (TLR2), IL-1ß and cyclooxygenase-2 (COX2) in hepatic tissues were detected. RESULTS: The lavandulyl flavonoid (kurarinol A, 1) and the EtOAc extract from S. flavescens showed protective effects on CCl4-injured HepG2 cells, increasing cell viability from 24.5% to 61.3% and 91.8%, respectively. What's more, we found that treatment with kurarinol A (1) and the EtOAc extract lead to a significant reduction in hepatotoxicity in response to acute CCl4 exposure. Compared with the model group, experimental results exhibited kurarinol A (10 mg/kg, i.p.) and the EtOAc extract (300 mg/kg, i.p.) could decrease the levels of AST, ALT, ALP and tissue damage. Further mechanistic investigations revealed that up-regulated the mRNA expression of SOD2, Nrf2, OH-1 and down-regulated the IL-1ß in liver tissues, respectively. Additionally, Western blot analyses elucidated that inhibition of IL-1ß, TLR2, COX-2, NF-κB (p65/p-p65) via TLR2/NF-κB signaling pathway by kurarinol A and the EtOAc extract contribute to its hepatoprotective activity. CONCLUSION: These findings demonstrated that the novel compound (kurarinol A, 1) possessed notable hepatoprotective activity against CCl4. It was confirmed that kurarinol A had a certain effect on mice with liver damage induced by CCl4, and its mechanism could be include inhibiting inflammation and reducing of oxidative stress reaction by regulating expression of related genes and proteins. Thus, kurarinol A could as a novel active agent that contributes to the hepatoprotective activity of S. flavescens for the treatment of live injury.

One-week phonemic training rebuilds the memory traces of merged phonemes in merged speakers.

The phonemic merger is a unique phenomenon which is referred to as acoustically very different phonemes are recognized as the same phoneme. In our previous study, we demonstrated that the merged speakers had lost the ability to discriminate the merged phonemes pre-attentively, as revealed by their failure in mismatch negativity (MMN) elicitation in the oddball stream of the merged phonemes /n/-/l/. In this study, we investigated the recovery of the discrimination ability via phonemic training and found that the merged speakers regained the ability of discriminating merged phonemes pre-attentively, after a 7-day /n/-/l/ phonemic training, as revealed by the reactivation of MMN brain response to the /n/-/l/ phoneme categories. Our finding indicates that separate memory traces of merged phonemes could be rebuilt during the training process.

Myristoleic acid produced by enterococci reduces obesity through brown adipose tissue activation.

OBJECTIVE: Dietary fibre has beneficial effects on energy metabolism, and the majority of studies have focused on short-chain fatty acids produced by gut microbiota. Ginseng has been reported to aid in body weight management, however, its mechanism of action is not yet clear. In this study, we focused on the potential modulating effect of ginseng on gut microbiota, aiming to identify specific strains and their metabolites, especially long-chain fatty acids (LCFA), which mediate the anti-obesity effects of ginseng. DESIGN: Db/db mice were gavaged with ginseng extract (GE) and the effects of GE on gut microbiota were evaluated using 16S rDNA-based high throughput sequencing. To confirm the candidate fatty acids, untargeted metabolomics analyses of the serum and medium samples were performed. RESULTS: We demonstrated that GE can induce Enterococcus faecalis, which can produce an unsaturated LCFA, myristoleic acid (MA). Our results indicate that E. faecalis and its metabolite MA can reduce adiposity by brown adipose tissue (BAT) activation and beige fat formation. In addition, the gene of E. faecalis encoding Acyl-CoA thioesterases (ACOTs) exhibited the biosynthetic potential to synthesise MA, as knockdown (KD) of the ACOT gene by CRISPR-dCas9 significantly reduced MA production. Furthermore, exogenous treatment with KD E. faecalis could not reproduce the beneficial effects of wild type E. faecalis, which work by augmenting the circulating MA levels. CONCLUSIONS: Our results demonstrated that the gut microbiota-LCFA-BAT axis plays an important role in host metabolism, which may provide a strategic advantage for the next generation of anti-obesity drug development.

Responses of water quality and phytoplankton assemblages to remediation projects in two hypereutrophic tributaries of Chaohu Lake.

Water shortages and the presence of point and diffuse source pollution have caused a serious deterioration in water quality in two tributaries (the Tangxi River and Shiwuli River) of Chaohu Lake, China. To reduce nutrient pollution and suppress harmful algal blooms (HABs), hard engineering and ecological remediation projects were implemented. A post-project investigation from 2013 to 2016 was carried out to evaluate the outcome of the remediation projects by monitoring the seasonal and spatial variations in water quality and the phytoplankton community. In the Tangxi River, the average total phosphorus (TP) concentrations in the four seasons were below 0.5 mg L-1, with the lowest concentration (0.29 ±â€¯0.12 mg L-1) found in autumn. Remediation measures including sediment dredging, riparian buffer zone creation, downstream wetland park construction, and water augmentation using reclaimed water and filtered lake water might combine to promote P source mitigation. Moreover, the percentage of bloom-forming cyanobacteria (i.e., Microcystis, Aphanizomenon, Anabaena, Oscillatoria, Phormidium and Planktothrix) in the phytoplankton assemblage and the biomass of the dominant species indicated successful HAB control. In the Shiwuli River, water quality improvements and phytoplankton responses have been observed since 2015 after the upgrading of a local wastewater treatment plant (WWTP) with effluent that was used for flow augmentation. Nevertheless, there is still room for improvement via increasing the river self-purification ability (e.g., the creation of downstream wetlands and riparian buffer zones) and promoting water augmentation according to the experience gained in the remediation projects of the Tangxi River.

[Status of diagnosis and management of acute appendicitis in 2017: a national multi-center retrospective study].

OBJECTIVE: To analyze the current status of diagnosis and management of acute appendicitis (AA) in China. METHODS: Questionnaire survey was used to retrospectively collect data of hospitalized patients with AA from 43 medical centers nationwide in 2017 (Sort by number of cases provided: Jinling Hospital of Medical School of Nanjing University, The First Affiliated Hospital of Xinjiang Medical University, Lu'an People's Hospital, Tengzhou Central People's Hospital, Dalian Central Hospital, The Affiliated Hospital of Xuzhou Medical University, Dongying People's Hospital, Jinjiang Hospital of Traditional Chinese Medicine, Huangshan Shoukang Hospital, Xuyi People's Hospital, Nanjing Jiangbei People's Hospital, Lanzhou 940th Hospital of PLA, Heze Municipal Hospital, The First College of Clinical Medical Science of China Three Gorges University, Affiliated Jiujiang Hospital of Nanchang University, The Second People's Hospital of Hefei, Affiliated Central Hospital of Shandong Zaozhuang Mining Group, The Third People's Hospital of Kunshan City, Xuzhou First People's Hospital, The 81st Group Army Hospital of PLA, Linyi Central Hospital, The General Hospital of Huainan Eastern Hospital Group, The 908th Hospital of PLA, Liyang People's Hospital, The 901th Hospital of Joint Logistic Support Force, The Third Affiliated Hospital of Chongqing Medical University, The Fourth Hospital of Jilin University, Harbin Acheng District People's Hospital, The First Affiliated Hospital of Zhengzhou University, Nanjing Luhe People's Hospital, Taixing Municipal People's Hospital, Baotou Central Hospital, The Affiliated Hospital of Nantong University, Linyi People's Hospital, The 72st Group Army Hospital of PLA, Zaozhuang Municipal Hospital, People's Hospital of Dayu County, Taixing City Hospital of Traditional Chinese Medicine, Suzhou Municipal Hospital, Beijing Guang'anmen Hospital, Langxi County Hospital of Traditional Chinese Medicine, Nanyang Central Hospital, The Affiliated People's Hospital of Inner Mongolia Medical University).The diagnosis and management of AA were analyzed through unified summary. Different centers collected and summarized their data in 2017 and sent back the questionnaires for summary. RESULTS: A total of 8 766 AA patients were enrolled from 43 medical centers, including 4 711 males (53.7%) with median age of 39 years and 958 (10.9%) patients over 65 years old. Of 8 776 patients, 5 677 cases (64.6%) received one or more imaging examinations, and the other 3 099 (35.4%) did not receive any imaging examination. A total of 1 858 (21.2%) cases received medical treatment, mainly a combination of nitroimidazoles (1 107 cases, 59.8%) doublet regimen, followed by a single-agent regimen of non-nitroimidazoles (451 cases, 24.4%), a nitroimidazole-free doublet regimen (134 cases, 7.2%), a triple regimen of combined nitroimidazoles (116 cases, 6.3%), nitroimidazole alone (39 cases, 2.1%) and nitroimidazole-free triple regimen (3 cases, 0.2%). Of the 6 908 patients (78.8%) who underwent surgery, 4 319 (62.5%) underwent laparoscopic appendectomy and 2589 (37.5%) underwent open surgery. Ratio of laparotomy was higher in those patients under 16 years old (392 cases) or over 65 years old (258 cases) [15.1%(392/2 589) and 10.0%(258/2 589), respectively, compared with 8.5%(367/4 316) and 8.0%(347/4 316) in the same age group for laparoscopic surgery, χ²=91.415, P<0.001; χ²=15.915,P<0.001]. Patients with complicated appendicitis had higher ratio of undergoing open surgery as compared to those undergoing laparoscopic surgery [26.7%(692/2 589) vs. 15.6%(672/4 316), χ²=125.726, P<0.001].The cure rates of laparoscopic and open surgery were 100.0% and 99.8%(2 585/2 589) respectively without significant difference (P=0.206). Postoperative complication rates were 4.5%(121/2 589) and 4.7%(196/4 316) respectively, and the difference was not statistically significant (χ²=0.065, P=0.799). The incidence of surgical site infection was lower (0.6% vs. 1.7%, χ²=17.315, P<0.001), and hospital stay was shorter [6(4-7) days vs. 6(5-8) days, U=4 384 348.0, P<0.001] in the laparoscopic surgery group, while hospitalization cost was higher (median 12 527 yuan vs. 9 342 yuan, U=2 586 809.0, P<0.001). CONCLUSIONS: The diagnosis of acute appendicitis is still clinically based, supplemented by imaging examination. Appendectomy is still the most effective treatment at present. Laparoscopic appendectomy has become the main treatment strategy, but anti-infective drugs are also very effective.

The use of vertical flow constructed wetlands for the treatment of hyper-eutrophic water bodies with dense cyanobacterial blooms.

Eutrophication often leads to the periodic proliferation of harmful cyanobacterial blooms (HCBs), which threaten the sustainability of freshwater ecosystems and lead to serious environmental, health and economic damage. Hence, it is vitally important to take effective measures to manage HCBs and associated problems. In this study, vertical flow constructed wetlands (CWs) were operated under different hydraulic loading rates (HLRs) to treat a hyper-eutrophic water body with HCBs. Six sampling ports (representing different layers) were evenly distributed along the water flow direction to study the purification processes of CWs. With HLRs ranging from 0.2 m/d to 0.8 m/d, total nitrogen (TN), total phosphorus (TP), COD, total suspended solid (TSS) and Chlorophyll a (Chl.a) were efficiently treated by CWs, and they were mainly removed at the second layer of CWs. The concentrations of two cyanobacterial metabolites (geosmin and ß-cyclocitral) in the effluent were mostly below their odorous threshold concentrations. As the HLRs increased, the treatment efficiencies of the CWs decreased gradually. There was no removal of TP, Chl.a, geosmin, or ß-cyclocitral at an HLR of 1.0 m/d. Under suitable HLRs, this type of CW could provide a promising way to control HCBs and associated odorous problems in hyper-eutrophic water bodies.

Enzyme Degradable Hyperbranched Polyphosphoester Micellar Nanomedicines for NIR Imaging-Guided Chemo-Photothermal Therapy of Drug-Resistant Cancers.

Multidrug resistance (MDR) is the major cause for chemotherapy failure, which constitutes a formidable challenge in the field of cancer therapy. The synergistic chemo-photothermal treatment has been reported to be a potential strategy to overcome MDR. In this work, rationally designed enzyme-degradable, hyperbranched polyphosphoester nanomedicines were developed for reversing MDR via the codelivery of doxorubicin and IR-780 (hPPEDOX&IR) as combined chemo-photothermal therapy. The amphiphilic hyperbranched polyphosphoesters with phosphate bond as the branching point were synthesized via a simple but robust one-step polycondensation reaction. The self-assembled hPPEDOX&IR exhibited good serum stability, sustained release, preferable tumor accumulation, and enhanced drug influx of doxorubicin in resistant MCF-7/ADR cells. Moreover, the degradation of hPPEDOX&IR was accelerated in the presence of alkaline phosphatase, which was overexpressed in various cancers, resulting in the fast release of encapsulated doxorubicin. The enzyme-degradable polymer generated synergistic chemo-photothermal cytotoxicity against MCF-7/ADR cells and, thus, the efficient ablation of DOX-resistant tumor without regrowth. This delivery system may open a new avenue for codelivery of chemo- and photothermal therapeutics for MDR tumor therapy.

Metabolite profiling of ginsenosides in rat plasma, urine and feces by LC-MS/MS and its application to a pharmacokinetic study after oral administration of Panax ginseng extract.

Panax ginseng is widely consumed as a functional food in the form of tea, powder, capsules, among others, and possesses a range of pharmacological activities including adaptogenic, immune-modulatory, anti-tumor, anti-aging and anti-inflammatory effects. The aim of this study was to identify and quantify the major ginsenosides and their metabolites in rat plasma, urine and feces after administration of P. ginseng extract using LC-MS/MS. We collected rat plasma samples at 0.5, 1, 2, 4, 8, 12, 24 and 48 h, and the amounts of urine and fecal samples accumulated in 24 h. Fourteen major ginsenosides and their metabolites were observed in fecal samples at high levels; however, low levels of 11 ginsenosides were detected in urine samples. The pharmacokinetics of the major ginsenosides and their metabolites was investigated in plasma. The results indicated that the maximum plasma concentration, time to maximum concentration and area under the curve of compound K were significantly greater than those of other ginsenosides. This study thus provides valuable information for drug development and clinical application of P. ginseng.

Comparative Analysis of the Rats' Gut Microbiota Composition in Animals with Different Ginsenosides Metabolizing Activity.

Following oral intake of Panax ginseng, major ginsenosides are metabolized to deglycosylated ginsenosides by gut microbiota before absorption into the blood. As the composition of gut microbiota varies between individuals, metabolic activities are significantly different. We selected 6 rats with low efficiency metabolism (LEM) and 6 rats with high efficiency metabolism (HEM) from 60 rats following oral administration of Panax ginseng extract, and analyzed their gut microbiota composition using Illumina HiSeq sequencing of the 16S rRNA gene. The components of gut microbiota between the LEM and HEM groups were significantly different. Between the 2 groups, S24-7, Alcaligenaceae, and Erysipelotrichaceae occupied most OTUs of the HEM group, which was notably higher than the LEM group. Furthermore, we isolated Bifidobacterium animalis GM1 that could convert the ginsenoside Rb1 to Rd. The result implies that these specific intestinal bacteria may dominate the metabolism of Panax ginseng.

Increasing phytoplankton-available phosphorus and inhibition of macrophyte on phytoplankton bloom.

We assembled mesocosms to address the coherent mechanisms that an increasing phosphorus (P) concentration in water columns coupled with the phytoplankton bloom and identify the performance gap of regulating phytoplankton growth between two macrophyte species, Ceratophyllum demersum L. and Vallisneria spiralis L. Intense alkaline phosphatase activities (APA) were observed in the unplanted control, with their predominant part, phytoplankton APA (accounting for up to 44.7% of the total APA), and another large share, bacterial APA. These correspond with the large average concentration of total phosphorus (TP), total dissolved phosphorus (TDP) and soluble reactive (SRP) as well as high phytoplankton density in the water column. The consistency among P concentrations, phytoplankton density and APA, together with the positive impact of phytoplankton density on total APA revealed by the structural equation modelling (SEM), indicates that facilitated APA levels in water is an essential strategy for phytoplankton to enhance the available P. Furthermore, a positive interaction between phytoplankton APA and bacteria APA was detected, suggesting a potential collaboration between phytoplankton and bacteria to boost available P content in the water column. Both macrophyte species had a prominent performance on regulating phytoplankton proliferation. The phytoplankton density and quantum yield in C. demersum systems were all significantly lower (33.8% and 24.0%) than those in V. spiralis systems. Additionally, a greater decoupling effect of C. demersum on the relationship between P, APA, phytoplankton density, bacteria dynamic and quantum yield was revealed by SEM. These results imply that the preferred tactic of different species could lead to the performance gap.

Biotransformation of gypenoside XVII to compound K by a recombinant ß-glucosidase.

OBJECTIVE: To study the ß-glucosidase gene (bgy1) from Lactobacillus brevis that was cloned and expressed in Escherichia coli BL21 (DE3) and then using it for the biotransformation of gypenoside XVII. RESULTS: The bgy1 gene consists of 2283 bp encoding 761 amino acids, with homology to the glycosyl hydrolase family-3 protein domain. The enzyme (Bgy1) hydrolyzed the glucose moieties at the C-3 position and the outer glucose moieties at the C-20 position of gypenoside XVII. Using 0.1 mg enzyme ml(-1) in 20 mM sodium phosphate buffer at 30 °C and pH 6.0, 1 mg gypenoside XVII ml(-1) was transformed into 0.58 mg compound K ml(-1) within 6 h, with a corresponding molar conversion yield of 89 %. CONCLUSION: The recombinant Bgy1 is considered potentially useful for the practical preparation of compound K.

Performance evaluation of wastewater treatment using horizontal subsurface flow constructed wetlands optimized by micro-aeration and substrate selection.

The effects of micro-aeration and substrate selection on domestic sewage treatment performance were explored using three pairs (with or without micro-aeration) of horizontal subsurface flow (HSSF) constructed wetlands (CWs) filled with zeolite, ceramsite or quartz granules. The individual and combined effects of micro-aeration and substrate selection on the purification performance of the experimental-scale HSSF CWs were evaluated. The results showed that micro-aeration significantly increased the treatment efficiencies for chemical oxygen demand, total nitrogen, total phosphorus (TP), ortho-phosphate (PO4(3-)-P) and ammonium nitrogen (NH4+-N) using HSSF CWs, while the substrate selection significantly affected the TP, PO4(3-)-P and NH4+-N removal efficiencies (p<0.05). A two-way analysis of variance (ANOVA) indicated that there was a significant interaction term (i.e. micro-aeration×substrate selection) for NH4+-N removal (p<0.05). Among the three substrates, ceramsite was the best substrate for the treatment of domestic sewage using HSSF CWs. Therefore, the results of this study suggest that a ceramsite-filled HSSF CW with micro-aeration could be the optimal configuration for decentralized domestic sewage treatment.

A field study on phytoremediation of dredged sediment contaminated by heavy metals and nutrients: the impacts of sediment aeration.

Compared to traditional chemical or physical treatments, phytoremediation has proved to be a cost-effective and environmentally sound alternative for remediation of contaminated dredged sediment. A field study was conducted in a sediment disposal site predominantly colonized by Typha angustifolia under different sediment moisture conditions to estimate the phytoremediation effects of dredged sediment. The moisture content was 37.30 % and 48.27 % in aerated and waterlogged sediment, respectively. Total nitrogen (TN) content was higher in the waterlogged sediment than in the aerated sediment. The total Cd contents were lower in aerated sediment, which was mainly resulted from the lower exchangeable fraction of Cd. The bioaccumulation of P, Cu and Pb in T. angustifolia was promoted by waterlogging, and the belowground tissue concentrations and accumulation factors (AFs) of Cu were higher than that of other metals, which can be explained by that Cu is an essential micronutrient for plants. Consistent with many previous studies, T. angustifolia showed higher metal levels in roots than in above-ground tissues at both the sediment conditions. Due to the improved biomass produced in the aerated sediment, the removals of nutrients and the metals by plant harvest were higher from aerated sediment than from waterlogged sediment. It was indicated that maintaining the dredged sediment aerated can avoid release risk and plant uptake of metals, while the opposite management option can promote phytoextraction of these contaminants.

Melatonin overcomes apoptosis resistance in human hepatocellular carcinoma by targeting survivin and XIAP.

Apoptosis resistance in hepatocellular carcinoma (HCC) is one of the most significant factors for hepatocarcinogenesis and tumor progression, and leads to resistance to conventional chemotherapy. It is well known that inhibitor of apoptosis proteins (IAPs) play key roles in apoptosis resistance, it has become an important target for antitumor therapy. In this study, we examined if melatonin, the main secretory product of the pineal gland, targeted IAPs, leading to the inhibition of apoptosis resistance. To accomplish this, we first observed that four members of IAPs (cIAP-1, cIAP-2, survivin, and XIAP) were overexpressed in human HCC tissue. Interestingly, melatonin significantly inhibited the growth of HepG2 and SMMC-7721 cells and promoted apoptosis along with the downregulation of survivin and XIAP, but had no effect on the expression of cIAP-1 and cIAP-2. These data suggest that the inhibition of survivin and XIAP by melatonin may play an important part in reversing apoptosis resistance. Notably, cIAP-1, survivin and XIAP were significantly associated with the coexpression of COX-2 in human HCC specimens. Melatonin also reduced the expression of COX-2 and inhibited AKT activation in HepG2 and SMMC-7721 cells. Inhibition of COX-2 activity with the selective inhibitor, NS398, and inhibition of AKT activation using the PI3K inhibitor, LY294002, in tumor cells confirmed that melatonin-induced apoptosis was COX-2/PI3K/AKT-dependent, suggesting that the COX-2/PI3K/AKT pathway plays a role in melatonin inhibition of IAPs. Taken together, these results suggest that melatonin overcomes apoptosis resistance by the suppressing survivin and XIAP via the COX-2/PI3K/AKT pathway in HCC cells.

Endoplasmic reticulum stress-induced resistance to doxorubicin is reversed by paeonol treatment in human hepatocellular carcinoma cells.

BACKGROUND: Endoplasmic reticulum stress (ER stress) is generally activated in solid tumors and results in tumor cell anti-apoptosis and drug resistance. Paeonol (Pae, 2-hydroxy-4-methoxyacetophenone), is a natural product extracted from the root of Paeonia Suffruticosa Andrew. Although Pae displays anti-neoplastic activity and increases the efficacy of chemotherapeutic drugs in various cell lines and in animal models, studies related to the effect of Pae on ER stress-induced resistance to chemotherapeutic agents in hepatocellular carcinoma (HCC) are poorly understood. METHODOLOGY/PRINCIPAL FINDINGS: In this study, we investigated the effect of the endoplasmic reticulum (ER) stress response during resistance of human hepatocellular carcinoma cells to doxorubicin. Treatment with the ER stress-inducer tunicamycin (TM) before the addition of doxorubicin reduced the rate of apoptosis induced by doxorubicin. Interestingly, co-pretreatment with tunicamycin and Pae significantly increased apoptosis induced by doxorubicin. Furthermore, induction of ER stress resulted in increasing expression of COX-2 concomitant with inactivation of Akt and up-regulation of the pro-apoptotic transcription factor CHOP (GADD153) in HepG2 cells. These cellular changes in gene expression and Akt activation may be an important resistance mechanism against doxorubicin in hepatocellular carcinoma cells undergoing ER stress. However, co-pretreatment with tunicamycin and Pae decreased the expression of COX-2 and levels of activation of Akt as well as increasing the levels of CHOP in HCC cells. CONCLUSIONS/SIGNIFICANCE: Our results demonstrate that Pae reverses ER stress-induced resistance to doxorubicin in human hepatocellular carcinoma cells by targeting COX-2 mediated inactivation of PI3K/AKT/CHOP.

Betaine attenuates Alzheimer-like pathological changes and memory deficits induced by homocysteine.

Hyperhomocysteinemia (Hhcy) may induce memory deficits with ß-amyloid (Aß) accumulation and tau hyperphosphorylation. Simultaneous supplement of folate and vitamin B12 partially restored the plasma homocysteine level and attenuated tau hyperphosphorylation, Aß accumulation and memory impairments induced by Hhcy. However, folate and vitamin B12 treatment have no effects on Hhcy which has the methylenetetrahydrofolate reductase genotype mutation. In this study, we investigated the effects of simultaneous supplement of betaine on Alzheimer-like pathological changes and memory deficits in hyperhomocysteinemic rats after a 2-week induction by vena caudalis injection of homocysteine (Hcy). We found that supplementation of betaine could ameliorate the Hcy-induced memory deficits, enhance long-term potentiation (LTP) and increase dendritic branches numbers and the density of the dendritic spines, with up-regulation of NR1, NR2A, synaptotagmin, synaptophysin, and phosphorylated synapsin I protein levels. Supplementation of betaine also attenuated the Hcy-induced tau hyperphosphorylation at multiple AD-related sites through activation protein phosphatase-2A (PP2A) with decreased inhibitory demethylated PP2A(C) at Leu309 and phosphorylated PP2A(C) at Tyr307. In addition, supplementation of betaine also decreased Aß production with decreased presenilin-1 protein levels. Our data suggest that betaine could be a promising candidate for arresting Hcy-induced AD-like pathological changes and memory deficits.

Case study on rehabilitation of a polluted urban water body in Yangtze River Basin.

In the past three decades, the fast development of economy and urbanization has caused increasingly severe pollutions of urban water bodies in China. Consequently, eutrophication and deterioration of aquatic ecosystem, which is especially significant for aquatic vegetation, inevitably became a pervasive problem across the Yangtze River Basin. To rehabilitate the degraded urban water bodies, vegetation replanting is an important issue to improve water quality and to rehabilitate ecosystem. As a case study, a representative polluted urban river, Nanfeihe River, in Hefei City, Anhui Province, was chosen to be a rehabilitation target. In October 2009 and May 2010, 13 species of indigenous and prevalent macrophytes, including seven species emergent, one species floating leaved, and five species submersed macrophytes, were planted along the bank slopes and in the river. Through 1.5 years' replanting practice, the water quality and biodiversity of the river had been improved. The concentrations of total nitrogen (TN), total phosphorus (TP), and ammonia nitrogen (NH4 (+)-N) declined by 46.0, 39.5, and 60.4 %, respectively. The species of macrophytes increased from 14 to 60, and the biodiversity of phytoplankton rose significantly in the river (p<0.05). The biomasses of zooplankton and benthos were also improved after the vegetation replanting. The study confirmed that vegetation replanting could alleviate the increasing water pollution and rehabilitate the degraded aquatic ecosystem. The case study would be an example for polluted urban waters restoration in the middle-downstream area of Yangtze River Base.

Four new N-contained iridoid glycosides from flower buds of Lonicera japonica.

Four new N-contained iridoid glycosides, lonijapospiroside A (1), L-phenylalaninosecologanin B (2), L-phenylalaninosecologanin C (3), and dehydroprolinoylloganin A (4), were isolated from the flower buds of Lonicera japonica Thunb. Their structures were established on the basis of UV, IR, MS, and NMR spectral data.

Four new flavonoids from Morus australis.

Four new flavonoids, benzokuwanon E (1), hydroxymorusin (2), dicyclokuwanon EA (3), and dicyclokuwanon EB (4), were isolated from Morus australis. Their structures were elucidated on the basis of UV, IR, MS, NMR, and CD spectral data.