Water-Soluble Total Flavonoids Isolated from Isodon lophanthoides var. gerardianus (Benth.) H. Hara Promote Hepatocellular Carcinoma Sensitivity to 5-Fluorouracil.

Isodon lophanthoides var. gerardianus (Benth.) H. Hara, a native medicinal plant produced chiefly across Southern China, is one of the mainstream varieties of Xihuangcao, which has long been applied in preventing and treating some common liver or gall diseases. Water-soluble total flavonoids (WSTF) extracted from folk herbal medicine have many pharmacological effects. The objective of the paper is to investigate the synergy of WSTF with 5-fluorouracil (5-FU) on HCC and the related mechanisms. Cells were exposed to WSTF alone or combination treatment with 5-FU. Then, in this study, we conducted cell viability test, cell cycle and clone forming test, apoptosis assay, reactive oxygen species (ROS), Western blotting, immunohistochemistry, and a xenograft tumor growth model for investigating the role of WSTF in HCC in vivo and in vitro. It was discovered that WSTF caused cell cycle arrest at the G0/G1 phase while increasing the ROS contents. The generation of ROS levels could cause cell apoptosis and inhibit colony formation. WSTF decreased the Bcl-2 level but promoted the Bax level. These showed the mitochondrial dependence of WSTF-mediated apoptosis. WSTF combined with 5-FU have a synergistic effect to significantly inhibit carcinogenicity in vivo and in vitro. The reduced ROS changed the synergy of WSTF with 5-FU. At last, WSTF inhibit the growth of HCC and promote the HCC sensitivity to 5-FU through ROS accumulation. WSTF-increased ROS levels may partially or completely contribute to enhanced toxicity. WSTF combined with 5-FU in HCC can play a synergistic effect when applied in the clinical setting.

Evaluation of the effects of the water-soluble total flavonoids from Isodon lophanthoides var.gerardianus (Benth.) H. Hara on apoptosis in HepG2 cell: Investigation of the most relevant mechanisms.

ETHNOPHARMACOLOGICAL RELEVANCE: The water-soluble total flavonoids (WSTF) were extracted from Isodon lophanthoides var. gerardianus (Benth.) H. Hara, a common folk herbal medicine in China, which has been recorded by the "Chinese Pharmacopoeia" in 2015 and used for prevention and clinical treatment of common diseases of liver and gall for many years. OBJECTIVE OF THE STUDY: The aim of this study is to evaluate the effects of WSTF on apoptosis in HepG2 cell and investigate the relevant mechanisms underlying. MATERIALS AND METHODS: Cytotoxicity was evaluated in HepG2 cells (human hepatoma cell lines) using MTT assay. The influence of the WSTF on the intracellular reactive oxygen species (iROS) and the mitochondrial membrane potential were also determinated. We used flow cytometry analysis to detect the effects of WSTF on apoptosis, cell cycle. Then we applied RT-PCR for genetic expression of main effectors and western blot analysis for activation of main effectors involved in the potential apoptosis signaling pathways. RESULTS: WSTF inhibited cell growth in HepG2 cells. Moreover, WSTF stimulates to increase amount of iROS, mitochondrial membrane potential, and the apoptotic relevant factors (cytochrome c, caspase-3) in HepG2 cells. WSTF could significantly induce apoptosis through downregulating apoptosis-antagonizing protein (Bcl-2, Survivin, mcl-1) and upregulating apoptosis-promoting proteins (Bax) and cell cycle G0/G1 arrest in HepG2 cells. CONCLUSION: The results indicate that WSTF induces cell apoptosis through mitochondrial pathway in the HepG2 cells. Therefore, these studies suggest that WSTF could be used as a chemotherapeutic agent to treat hepatoma.

[Microbial diversity and ammonia-oxidizing microorganism of a soil sample near an acid mine drainage lake].

The main physicochemical parameters of the soil sample which was collected near an acid mine drainage reservoir in Anhui province was analyzed. The microbial diversity and community structure was studied through the construction of bacteria and archaea 16S rRNA gene clone libraries and ammonia monooxygenase gene clone library of archaea. The functional groups which were responsible for the process of ammonia oxidation were also discussed. The results indicated that the soil sample had extreme low pH value (pH < 3) and high ions concentration, which was influenced by the acid mine drainage (AMD). All the 16S rRNA gene sequences of bacteria clone library fell into 11 phyla, and Acidobacteria played the most significant role in the ecosystem followed by Verrucomicrobia. A great number of acidophilic bacteria existed in the soil sample, such as Candidatus Koribacter versatilis and Holophaga sp.. The archaea clone library consisted of 2 phyla (Thaumarchaeota and Euryarchaeota). The abundance of Thaumarchaeota was remarkably higher than Euryarchaeota. The ammonia oxidation in the soil environment was probably driven by ammonia-oxidizing archaea, and new species of ammonia-oxidizing archaea existed in the soil sample.

[Microeukaryotic biodiversity in the waste ore samples surrounding an acid mine drainage lake].

The abandoned mineral samples were collected in an acid mine drainage area in Anhui Province. Molecular ecological methods were used to construct 18S rDNA clone libraries after analyzing the main physicochemical parameters, and then the microeukaryotic diversity and community structure in the acid mine drainage area were studied. The results showed that the region was strongly acidic (pH <3), and the concentrations of Fe, SO2-(4), P, NO-(3) -N showed the same trend, all higher in the bare waste ore samples PD and 1 M than in the vegetation covered samples LW and XC. Four eukaryotic phyla were detected in the abandoned mineral samples: Ascomycota, Basidiomycota, Glomeromycota and Arthropoda. Glomeromycota can form an absolute symbiotic relationship with the plant, and it was a key factor for early plant to adapt the terrestrial environment. The biodiversity of the vegetation covered samples LW and XC, which contained Glomeromycota, was much higher than that of the bare abandoned rock samples PD and 1 M. Moreover, many sequences in the libraries were closely related to some isolated strains, which are tolerant to low pH and heavy metals, such as Penicillium purpurogenum, Chaetothyriales sp. and Staninwardia suttonii.

[Performance of nitrogen and phosphorus removal from municipal wastewater of different C/N ratios using intelligent controlled systems sequencing batch biofilm reactor ( SBBR)].

The laboratory-scale experiments were conducted to treat synthetic municipal wastewater at different C/N ratios, using a developed intelligent controller sequencing batch biofilm reactor (SBBR), which formed alternative aerobic-anoxic environment. The performance of nitrogen and phosphorus removal was investigated under different conditions (T = 25 degrees C +/- 1 degrees C, V(aeration) = 150 L/h, COD = 300 mg/L, TP = 5 mg/L, TN = 30 mg/L, t(operation) = 15 d; TN = 60 mg/L, t(operation) = 20 d; TN = 90 mg/L, t(operation) = 20 d). The average removal efficiency of COD reaches 85.2%, 91.1% and 97.7%, the average removal efficiency of TP achieves 94.1%, 95.9%, 96.7% ,the average removal efficiency of NH4(+) -N reaches 95.8%, 99.2%, 80.0%, and the average removal efficiency of TN are 90.7%, 81.1%, 58.3%, respectively. With the decrease of C/N ratios, the removal efficiency of N decreases significantly, while the removal efficiency of COD and TP increase slightly. In intelligent controlled SBBR, simultaneous nitrification and denitrification and shortened simultaneous nitrification and denitrification occur, meanwhile, the denitrification of the system strengthens with the increasing of C/N ratios. After the influent finished, intelligent controlled SBBR starts to operate in aerobic-anoxic environment. The efficiency of phosphorus removal is high, moreover, the theory of phosphorus removal is different from the conventional theory of release anaerobic and uptake aerobic phosphorus.