[Removal of Oxytetracycline from Water Using Blast Furnace Slag Loaded Sulfide Nanoscale Zero-valent Iron].

Blast furnace slag loaded with sulfide nano zero valent iron (S-nZVI@BFS) was applied to remove oxytetracycline (OTC) from water. S-nZVI@BFS was synthesized via liquid reduction and characterized using scanning electron microscopy (SEM), X-ray diffraction (XRD), and Brunauer, Emmett and Teller (BET) theory. The effect of reaction time, initial concentration of OTC, initial pH, and coexisting hexavalent chromium[Cr(Ⅵ)] were investigated. The results show that Fe and S were successfully immobilized on the surface of S-nZVI@BFS, the specific surface area and pore volume of which increased to 141.986 m2·g-1 and 0.388 cm3·g-1, respectively, following the loading of nZVI and sulfurization. The utilization rate of the surface active sites of S-nZVI@BFS was improved with an increase of the initial concentration of OTC; the removal rate increased from 20.12 mg·g-1 to 202.74 mg·g-1 when the initial concentration of OTC was increased from 10 mg·L-1 to 100 mg·L-1. The removal rate decreased with pH, declining from 99.78 mg·g-1 to 41.12 mg·g-1 when pH was increased from 3 to 11 due to the switch from Fendon oxidation to electrostatic adsorption. There was notable competition between OTC and Cr(Ⅵ) meaning that Cr(Ⅵ) can inhibit the removal of OTC, which is dose dependent.

Signal transducer and activator of transcription 3 promotes the Warburg effect possibly by inducing pyruvate kinase M2 phosphorylation in liver precancerous lesions.

BACKGROUND: Study shows that signal transducer and activator of transcription 3 (STAT3) can increase the Warburg effect by stimulating hexokinase 2 in breast cancer and upregulate lactate dehydrogenase A and pyruvate dehydrogenase kinase 1 in myeloma. STAT3 and pyruvate kinase M2 (PKM2) can also be activated and enhance the Warburg effect in hepatocellular carcinoma. Precancerous lesions are critical to human and rodent hepatocarcinogenesis. However, the underlying molecular mechanism for the development of liver precancerous lesions remains unknown. We hypothesized that STAT3 promotes the Warburg effect possibly by upregulating p-PKM2 in liver precancerous lesions in rats. AIM: To investigate the mechanism of the Warburg effect in liver precancerous lesions in rats. METHODS: A model of liver precancerous lesions was established by a modified Solt-Farber method. The liver pathological changes were observed by HE staining and immunohistochemistry. The transformation of WB-F344 cells induced with N-methyl-N'-nitro-N-nitrosoguanidine and hydrogen peroxide was evaluated by the soft agar assay and aneuploidy. The levels of glucose and lactate in the tissue and culture medium were detected with a spectrophotometer. The protein levels of glutathione S-transferase-π, proliferating cell nuclear antigen (PCNA), STAT3, and PKM2 were examined by Western blot and immunofluorescence. RESULTS: We found that the Warburg effect was increased in liver precancerous lesions in rats. PKM2 and p-STAT3 were upregulated in activated oval cells in liver precancerous lesions in rats. The Warburg effect, p-PKM2, and p-STAT3 expression were also increased in transformed WB-F344 cells. STAT3 activation promoted the clonal formation rate, aneuploidy, alpha-fetoprotein expression, PCNA expression, G1/S phase transition, the Warburg effect, PKM2 phosphorylation, and nuclear translocation in transformed WB-F344 cells. Moreover, the Warburg effect was inhibited by stattic, a specific inhibitor of STAT3, and further reduced in transformed WB-F344 cells after the intervention for PKM2. CONCLUSION: The Warburg effect is initiated in liver precancerous lesions in rats. STAT3 activation promotes the Warburg effect by enhancing the phosphorylation of PKM2 in transformed WB-F344 cells.

[Removal of Lead Ions from Water by Struvite Natural Zeolite Composite].

A nitrogen and phosphorus recovery product (NZ-MAP) containing struvite was applied to remove heavy metal ions and lead from water. NZ-MAP was characterized using XRD, FTIR, and SEM/EDS. The effects of dosage, initial pH, and reaction time on the removal process were investigated. The results show that the main component of NZ-MAP is a natural zeolite loaded with struvite. When the dosage is 0.4 g·L-1, the maximum adsorption amount is 749.74 mg·g-1, and the adsorption of Pb2+ in the solution increases first and then tends to reach a balance with increase of the pH. Its removal mechanism is mainly Pb10 (PO4)6 (OH)2 precipitation, and the effect is best when pH is 5.0. The removal process of Pb2+ from water is more in line with the quasi-two stage kinetic model. In an investigation of the effect of coexisting heavy metal ions on the removal of lead ions in water by NZ-MAP, it was found that coexisting Ni2+ and Cu2+ have little effect on NZ-MAP adsorption on Pb2+. Coexisting Zn2+ and Al3+ obviously inhibit the adsorption of Pb2+ by NZ-MAP. The research shows that NZ-MAP material can remove lead ions from water efficiently, thus offering an effective method for doing this.

[Simultaneous Recovery of Nutrients from Wastewater by Mesoporous MgO-loaded Natural Zeolital].

Using MgO modified natural zeolite (NZ-MgO) as a settling agent, nutrients from synthetic contaminated waters were recovered as a struvite precipitate. Different reaction conditions, including reagent dosage, solution pH, contact time, and coexisting calcium ions, were investigated. In order to understand the recovery mechanism, Fourier transform infrared spectroscopy (FTIR) and X-ray diffraction (XRD) were utilized to analyze the chemical composition of recovered precipitates, and Brunauer-Emmett-Teller (BET) and field-emission scanning electron microscopy(SEM) were used to detect the surface area and morphology. The results showed that NZ-MgO exhibited a strong removal capacity, with a maximum removal capacity of 119.2 mg·g-1 for phosphate and 48.5 mg·g-1 for ammonium nitrogen at a dosage of 0.4 g·L-1, initial pH of 7, and contact time of 2 h. The kinetics of both phosphate and ammonia nitrogen followed the pseudo-second-order kinetic model (R2>0.99), which revealed that the chemical process occurred at the solid-liquid interface. The mechanisms for nutrient recovery by NZ-MgO were struvite crystallization, physical adsorption, ion exchange, and electrostatic attraction, with struvite crystallization being the main mechanism. Existing calcium ions would hinder the simultaneous recovery of nutrients from wastewater, resulting in a struvite decline and calcium phosphate increase.

The effects of urotensin II on migration and invasion are mediated by NADPH oxidase-derived reactive oxygen species through the c-Jun N-terminal kinase pathway in human hepatoma cells.

AIMS: Urotensin II (UII) is a vasoactive neuropeptide involved in migration and invasion in various cell types. However, the effects of UII on human hepatoma cells still remain unclear. The aim of this study was to investigate the role and mechanism of UII on migration and invasion in human hepatoma cells. METHODS: Migration was measured by wound healing assays and a Transwell® methodology, and invasion was analyzed using Matrigel® invasion chambers. Reactive oxygen species (ROS) levels were detected using a 2', 7'-dichlorofluorescein diacetate probe, and flow cytometry, and protein expression levels were evaluated by western blotting. Cell proliferation and actin polymerization were examined using cell proliferation reagent WST-1 and F-actin immunohistochemistry staining. RESULTS: Exposure to UII promoted migration and invasion in hepatoma cells compared with that in cells without UII. UII also increased matrix metalloproteinase-2 (MMP2) expression in a time-independent manner. Furthermore, UII markedly enhanced ROS generation and NADPH oxidase subunit expression, and consequently facilitated the phosphorylation of c-Jun N-terminal kinase (JNK). The UT antagonist urantide or the antioxidant/NADPH oxidase inhibitor apocynin decreased UII-induced ROS production. JNK phosphorylation, migration, invasion, and MMP9/2 expression were also reversed by pretreatment with apocynin. Urantide and JNK inhibitor SP600125 abrogated migration, invasion, or MMP9/2 expression in response to UII. UII induced actin polymerization and fascin protein expression, and could be reversed by apocynin and SP600125. CONCLUSIONS: Exogenous UII induced migration and invasion in hepatoma cells that mainly involved NADPH oxidase-derived ROS through JNK activation. UT played an additional role in regulating hepatoma cells migration and invasion. Thus, our data suggested an important effect of UII in hepatocellular carcinoma metastasis.

Urotensin II-induced insulin resistance is mediated by NADPH oxidase-derived reactive oxygen species in HepG2 cells.

AIM: To investigated the effects of urotensin II (UII) on hepatic insulin resistance in HepG2 cells and the potential mechanisms involved. METHODS: Human hepatoma HepG2 cells were cultured with or without exogenous UII for 24 h, in the presence or absence of 100 nmol/L insulin for the last 30 min. Glucose levels were detected by the glucose-oxidase method and glycogen synthesis was analyzed by glycogen colorimetric/fluorometric assay. Reactive oxygen species (ROS) levels were detected with a multimode reader using a 2',7'-dichlorofluorescein diacetate probe. The protein expression and phosphorylation levels of c-Jun N-terminal kinase (JNK), insulin signal essential molecules such as insulin receptor substrate -1 (IRS-1), protein kinase B (Akt), glycogen synthase kinase-3ß (GSK-3ß), and glucose transporter-2 (Glut 2), and NADPH oxidase subunits such as gp91(phox), p67(phox), p47(phox), p40(phox), and p22(phox) were evaluated by Western blot. RESULTS: Exposure to 100 nmol/L UII reduced the insulin-induced glucose consumption (P < 0.05) and glycogen content (P < 0.01) in HepG2 cells compared with cells without UII. UII also abolished insulin-stimulated protein expression (P < 0.01) and phosphorylation of IRS-1 (P < 0.05), associated with down-regulation of Akt (P < 0.05) and GSK-3ß (P < 0.05) phosphorylation levels, and the expression of Glut 2 (P < 0.001), indicating an insulin-resistance state in HepG2 cells. Furthermore, UII enhanced the phosphorylation of JNK (P < 0.05), while the activity of JNK, insulin signaling, such as total protein of IRS-1 (P < 0.001), phosphorylation of IRS-1 (P < 0.001) and GSK-3ß (P < 0.05), and glycogen synthesis (P < 0.001) could be reversed by pretreatment with the JNK inhibitor SP600125. Besides, UII markedly improved ROS generation (P < 0.05) and NADPH oxidase subunit expression (P < 0.05). However, the antioxidant/NADPH oxidase inhibitor apocynin could decrease UII-induced ROS production (P < 0.05), JNK phosphorylation (P < 0.05), and insulin resistance (P < 0.05) in HepG2 cells. CONCLUSION: UII induces insulin resistance, and this can be reversed by JNK inhibitor SP600125 and antioxidant/NADPH oxidase inhibitor apocynin targeting the insulin signaling pathway in HepG2 cells.

Up-regulation of urotensin II and its receptor contributes to human hepatocellular carcinoma growth via activation of the PKC, ERK1/2, and p38 MAPK signaling pathways.

Urotensin II (UII) and its receptor (UTR) have mitogenic effects on tumor growth. Our previous study demonstrated that the UII/UTR system is up-regulated in dithyinitrosamine-induced precancerous rat liver lesions. However, its role in human hepatocellular carcinoma remains unknown. In this study, the mRNA and protein expression of UII and its receptor (UTR) in human hepatocellular carcinoma samples and in the BEL-7402 human hepatoma cell line were evaluated. In addition, the effect of exogenous UII on the pathways that regulate proliferation in BEL-7402 cells in vitro were determined. Liver sections were subjected to immunohistochemical staining. mRNA expression was detected by real-time polymerase chain reaction analysis, and protein levels were evaluated by western blotting. Proliferating cells were detected by BrdU incorporation. The expression of UII/UT mRNA and protein significantly increased in human hepatocellular carcinoma samples, and in BEL-7402 cells. Administration with UII increased the phosphorylation of protein kinase C (PKC), extracellular signal-regulated kinase (ERK1/2) and p38 mitogen-activated protein kinases (p38 MAPK). Furthermore, GF109203x, PD184352, and SB203580 partially abolished UII-induced proliferation of BEL-7402 cells. These results provide the first evidence that up-regulation of the UII/UT system may enhance proliferation of the human hepatoma cell line at least in part via PKC, ERK1/2, and p38 MAPK signaling pathways, and may provide novel therapeutic targets for inhibiting human hepatocellular carcinoma.

Mistletoe alkaloid fractions alleviates carbon tetrachloride-induced liver fibrosis through inhibition of hepatic stellate cell activation via TGF-ß/Smad interference.

ETHNOPHARMACOLOGICAL RELEVANCE: Mistletoe (Viscum coloratum (Kom.) Nakai) has long been categorized as a traditional herbal medicine in Asia. In addition to its application in cancer therapy, mistletoe has also been used in the treatment of chronic hepatic disorders in China. In the present study, we investigated the antifibrotic effect and mechanisms of action of mistletoe extracts in a rat model of carbon tetrachloride (CCl4)-induced hepatotoxicity. MATERIALS AND METHODS: An experimental model of hepatic fibrosis was established by intraperitoneal injection of rats with CCl4 for 8 weeks. Rats were subsequently treated with a mistletoe alkaloid fraction preparation via oral administration (120mg/kg daily for 8 weeks) or with distilled water as a control. Histopathological changes were observed by hematoxylin and eosin staining and Masson׳s trichrome staining. The expression of markers relevant to hepatic stellate cell (HSC) activation in the liver was assessed by real-time reverse transcription-polymerase chain reaction, immunohistochemistry and western blotting. The anti-fibrosis activity and mechanisms of action of mistletoe alkaloid fractions were further investigated in the HSC-T6 HSC line, following treatment with mistletoe alkaloid fractions (12mg/ml) for 48h. RESULTS: Hepatic fibrosis decreased markedly in CCl4-treated animals following treatment with mistletoe alkaloid fractions, compared to controls. The mRNA levels of transforming growth factor-ß1 (TGF-ß1), procollagen I and tissue inhibitors of metalloproteinases (TIMPs) were significantly downregulated, by about 40%, 40% and 45%, respectively, in liver tissues from rats treated with mistletoe alkaloid fractions. Furthermore, significant downregulation of TGF-ß1, TGF-ß1 receptor, phosphorylated Smad 2 and alpha smooth muscle actin (α-SMA) proteins, by about 45%, 30% and 40%, respectively, was also observed in liver tissues from mistletoe alkaloid fractions-treated rats. In contrast, Smad 7 levels were significantly increased by about 30% in mistletoe alkaloid fractions-treated rats. Treatment of HSC-T6 cells with mistletoe alkaloid fractions significantly induced Smad 7 expression and inhibited the expression of α-SMA, TGFß1, TGF-ß1 receptor, Smad 2 and TIMP-1, in vitro. CONCLUSION: We demonstrate that mistletoe alkaloid fractions decrease extracellular matrix accumulation by inhibiting HSC activation. Mechanistically, this may occur via inhibition of TGF-ß1/Smad 2 and Smad 7 signal transduction, thereby blocking the synthesis of procollagen I and TIMP-1. These findings suggest that mistletoe alkaloid fractions may be a potential therapeutic agent for the treatment of hepatic fibrosis.

[Characteristics of hydroxyapatite/PMMA nanocomposites for provisional restoration and its biocompatibility with human gingival fibroblasts].

OBJECTIVE: To formulate hydroxyapatite (HA)/polymethyl methacrylate (PMMA) composites with improved cytocompatibility for provisional restoration. METHODS: Nanocomposites with 20 wt%, 30 wt%, 40 wt%, and 50 wt% HA/PMMA (H/P) were developed and examined using X-ray photoelectron spectroscopy (XPS) and scanning electron microscope (SEM). Human gingival fibroblasts were cultured on those HA/PMMA discs and investigated by fluorescent staining on 24 h and MTT assay at 1 d, 3 d, 5 d and 7 d. RESULTS: Chemical integration of HA/PMMA interface was confirmed by XPS. Typical fusiform cells with adhesion spots were detected on 40 wt% and 50 wt% H/P discs. MTT results showed insignificant differences in cell growth between 40 wt% H/P and pure titanium (Ti, P > 0.05), while the other H/P discs showed significantly lower cell growth than pure Ti (P < 0.05). CONCLUSION: 40 wt% H/P might be a promising candidate for provisional dental implant restoration and for esthetic gingival contour.

Oleanolic acid and ursolic acid inhibit proliferation in transformed rat hepatic oval cells.

AIM: To investigate H2O2-induced promotion proliferation and malignant transformation in WB-F344 cells and anti-tumor effects of ursolic acid (UA) and oleanolic acid (OA). METHODS: WB-F344 cells were continuously exposed to 7 x 10(-7) mol/L H2O2 for 21 d. Observations of cell morphology, colony formation rates, flow cytometric analysis of cell cycle changes and aneuploidy formation indicated that H2O2 was able to induce malignant transformation of WB-F344 cells. We treated malignantly transformed WB-F344 cells with 4 µmol/L OA or 8 µmol/L UA for 72 h and analyzed the cell cycle distribution by flow cytometry. RESULTS: MTT assay showed that 7 x 10(-7) mol/L H2O2 decreased G1 phase subpopulation from 73.8% to 49.6% compared with the control group, and increased S phase subpopulation from 14.5% to 31.8% (P < 0.05 vs control group). Cell morphology showed that nucleus to cytoplasm ratio increased, many mitotic cells, prokaryotes and even tumor giant cells were shown in H2O2-induced WB-F344 cells. Fluorescence activated cell sorting analysis showed that WB-F344 cell aneuploidy increased to 12% following H2O2 treatment. Flow cytometric analysis of the transformed WB-F344 cells following treatment with OA (4 µmol/L) and UA (8 µmol/L) showed that OA increased G1 subpopulation to 68.6%, compared to 49.7% in unexposed cells. UA increased G1 subpopulation to 67.4% compared to 49.7% in unexposed cells (P < 0.05 vs H2O2 model group). CONCLUSION: H2O2 causes the malignant transformation of WB-F344 cells. OA and UA exert anti-tumor effects by inhibiting the proliferation in malignantly transformed WB-F344 cells.

Urotensin II inhibits skeletal muscle glucose transport signaling pathways via the NADPH oxidase pathway.

Our previous studies have demonstrated that the urotensin (UII) and its receptor are up-regulated in the skeletal muscle of mice with type II diabetes mellitus (T2DM), but the significance of UII in skeletal muscle insulin resistance remains unknown. The purpose of this study was to investigate the effect of UII on NADPH oxidase and glucose transport signaling pathways in the skeletal muscle of mice with T2DM and in C2C12 mouse myotube cells. KK/upj-AY/J mice (KK) mice were divided into the following groups: KK group, with saline treatment for 2 weeks; KK+ urantide group, with daily 30 µg/kg body weight injections over the same time period of urantide, a potent urotensin II antagonist peptide; Non-diabetic C57BL/6J mice were used as normal controls. After urantide treatment, mice were subjected to an intraperitoneal glucose tolerance test, in addition to measurements of the levels of ROS, NADPH oxidase and the phosphorylated AKT, PKC and ERK. C2C12 cells were incubated with serum-free DMEM for 24 hours before conducting the experiments, and then administrated with 100 nM UII for 2 hours or 24 hours. Urantide treatment improved glucose tolerance, decreased the translocation of the NADPH subunits p40-phox and p47-phox, and increased levels of the phosphorylated PKC, AKT and ERK. In contrast, UII treatment increased ROS production and p47-phox and p67-phox translocation, and decreased the phosphorylated AKT, ERK1/2 and p38MAPK; Apocynin abrogated this effect. In conclusion, UII increased ROS production by NADPH oxidase, leading to the inhibition of signaling pathways involving glucose transport, such as AKT/PKC/ERK. Our data imply a role for UII at the molecular level in glucose homeostasis, and possibly in skeletal muscle insulin resistance in T2DM.

Activation of apoptosis in hepatocellular carcinoma by the Chinese traditional medicine Hu Qisan.

To investigate the effects of Hu Qisan (HQS) on apoptosis in diethylnitrosamine (DEN)-induced hepatocellular carcinoma (HCC), a Solt-Farber two-step test model of precancerous liver lesions was established in rats using a previously described method. HQS (4 and 8 g/kg body weight/day) was administered for 4 weeks, after the majority of the liver was removed. HepG2 cells were used to detect the HtrA serine peptidase 2 (HtrA2/Omi) release from mitochondria and caspase-3 activation promoted by HQS. Exposure of the rats to DEN for 6 weeks induced hepatic carcinogenesis. HQS (4 and 8 g/kg body weight/day) markedly induced cell apoptosis. The protective effects against hepatic carcinogenesis were mediated by multiple mechanisms, including the reduction of DEN-induced γ-GT-positive cell proliferation, mitochondrial morphological changes, HtrA2/Omi release from mitochondria and the activation of caspase-3. In conclusion, HQS is a potential anti-carcinogenic agent that may induce apoptosis by reducing the inhibitory effects of X-linked inhibitor of apoptosis protein (XIAP) on caspase-3. Thus, HQS should be further explored as a potentially promising new therapeutic agent against human hepatic cancer.

[Copper and cadmium toxicities to activated sludge investigated with ToxTell biosensor].

Effects of different concentrations of Cu2+, Cu2+ and Cd2+ combined pollution on the removal performance of COD in activated sludge system were investigated, and the ToxTell biosensor with activated sludge was constructed to determine the toxicity of Cu2+, Cu2+ and Cd2+ combined pollution. The results showed that there was no significant impact on the activated sludge process when Cu2+ concentration was lower than 10 mg x L(-1), and the addition of Cd2+ enhanced the inhibition of COD removal, and the maximum inhibition efficiency of COD reached at about 1.5 h. With the increase of aeration time, the COD removal efficiency increased slowly again. The toxicity measurement with ToxTell biosensor was close to the biological effects (inhibition efficiencies of COD), which showed that the ToxTell biosensor could be used well in the early warming determination of Cu2+ and Cd2+ in the activated sludge process.

[Acute toxicity analysis performance of CellSense biosensor with E. coli].

E. coli microbial electrodes for CellSense biosensor were prepared by polycarbonate membrane immobilization process, and their performance for heavy metals and toxic organic compounds acute toxicity determination were studied. The results showed that when E. coli was in logarithmic and stationary phase, the CellSense biosensor with E. coli showed good performance in heavy metal ions and organic pollutants acute toxicity analysis, when E. coli was in its decline phase, the stability and sensitivity of the CellSense biosensor was poor. The EC50 values of Hg2+, Cu2+, Zn2+, o-chlorophenol (2-CP) and p-nitrophenol (4-NP) detected by CellSense biosensor with E. coli were 0.6, 3.1, 5.8, 180 and 94 microg/mL, respectively. The immobilized E. coli electrodes could still suit for acute toxicity assessment after 2 months storage at 4 degrees C.

[Flocculation characteristics of the return sludge in chemical-biological flocculation process].

The chemical-biological flocculation (CBF) process was used to treat municipal wastewater in Shanghai, and the effect of its return sludge on pollutant removal was studied through stopping chemical addition, simulating different return sludge ratio, and analyzing extracellular polymeric substances (EPS). The results show that the return sludge in CBF exhibits strong pollutant removal capability, and chemical in its return sludge can further TP removal and enhance CBF's adaptability to the influent TP impulse. After stopping PAFC (Al2 O3 : 10.8%, Fe2 O3: 1.8%) addition, COD removal efficiency is achieved at about 50%, and removal rates of PO4(3-) and TP gradually decline. It is also found that chemical addition plays an important role in promote sludge sedimentation in CBF. EPS extracted from CBF is 145.89 mg/g, while EPS from CEPT is only 17.24 mg/g, which indicates that there is strong biological flocculation behavior in CBF. With the increase of return sludge ratio, TP and COD removal efficiencies decrease in CEPT, which reveals that chemical in waste sludge has poor flocculation capability, while CBF return sludge exhibits good flocculation behavior due to its biological flocculation. Chemical flocculation and biological flocculation work collaboratively in CBF process.

[Toxicity assessments of landfill leachate based on CellSense biosensor].

The toxicity of landfill leachate of domestic wastes in different treatment unites landfill leachate pretreatment (No.1), facultative lagoon (No.2), aerated lagoon (No.3), mineralized refuse bioreactor (No.4) and catalysis iron inner electrolysis (No.5) of leachate treatment process in Shanghai was determined based on CellSense biosensor with E. coli. The results showed that the toxicity of the effluent in different treatment unites for landfill leachate increased during the treatment process due to the influence of matrix effect and intermediate toxic products of refractory organic matters. The toxicity of the effluent from No.1 to No.5 unites are 90% (EC10), 80% (EC30), 60% (EC50), 80% (EC50) and 2% (EC50), respectively. This showed no significant correlation with their corresponding COD and ammonia nitrogen concentrations.

Production and application of a novel bioflocculant by multiple-microorganism consortia using brewery wastewater as carbon source.

The flocculating activity of a novel bioflocculant MMF1 produced by multiple-microorganism consortia MM1 was investigated. MM1 was composed of strain BAFRT4 identified as Staphylococcus sp. and strain CYGS1 identified as Pseudomonas sp. The flocculating activity of MMF1 isolated from the screening medium was 82.9%, which is remarkably higher than that of the bioflocculant produced by either of the strains under the same condition. Brewery wastewater was also used as the carbon source for MM1, and the cost-effective production medium for MM1 mainly comprised 1.0 L brewery water (chemical oxygen demand (COD) 5000 mg/L), 0.5 g/L urea, 0.5 g/L yeast extract, and 0.2 g/L (NH4)2SO4. The optimal conditions for the production of MMF1 was inoculum size 2%, initial pH 6.0, cultivating temperature 30 degrees C, and shaking speed 160 r/min, under which the flocculating activity of the MMF1 reached 96.8%. Fifteen grams of purified bioflocculant could be recovered from 1.0 L of fermentation broth. MMF1 was identified as a macromolecular substance containing both protein and polysaccharide. It showed good flocculating performance in treating indigotin printing and dyeing wastewater, and the maximal removal efficiencies of COD and chroma were 79.2% and 86.5%, respectively.

Effect of Qi-protecting powder (Huqi San) on expression of c-jun, c-fos and c-myc in diethylnitrosamine-mediated hepatocarcinogenesis.

AIM: To study the inhibitory effect of Huqi San (Qi-protecting powder) on rat prehepatocarcinoma induced by diethylinitrosamine (DEN) by analyzing the mutational activation of c-fos proto-oncogene and over-expression of c-jun and c-myc oncogenes. METHODS: A Solt-Farber two-step test model of prehepatocarcinoma was induced in rats by DEN and 2-acetylaminofluorene (AAF) to investigate the modifying effects of Huqi San on the expression of c-jun, c-fos and c-myc in DEN-mediated hepatocarcinogenesis. Huqi San was made of eight medicinal herbs containing glycoprival granules, in which each milliliter contains 0.38 g crude drugs. gamma-glutamy-transpeptidase-isoenzyme (gamma-GTase) was determined with histochemical methods. Level of 8-hydroxydeoxyguanosine (OHdG) formed in liver and c-jun, c-fos and c-myc proto-oncogenes were detected by immunohistochemical methods. RESULTS: The level of 8-OHdG, a mark of oxidative DNA damage, was significantly decreased in the liver of rats with prehepatocarcinoma induced by DEN who received 8 g/kg body weight or 4 g/kg body weight Huqi San before (1 wk) and after DEN exposure (4 wk). Huqi San-treated rats showed a significant decrease in number of gamma-GT positive foci (P < 0.001, prevention group: 4.96 +/- 0.72 vs 29.46 +/- 2.17; large dose therapeutic group: 7.53 +/- 0.88 vs 29.46 +/- 2.17). On the other hand, significant changes in expression of c-jun, c-fos and c-myc were found in Huqi San-treated rats. CONCLUSION: Activation of c-jun, c-fos and c-myc plays a crucial role in the pathogenesis of liver cancer. Huqi San can inhibit the over-expression of c-jun, c-fos and c-myc oncogenes and liver preneolastic lesions induced by DEN.

[Analysis on mechanism of the chemical-biological flocculation process].

Zeta potential, particle size distribution and molecular weight distribution of dissolved TOC were studied to elementarily disclose the mechanism of the chemical-biological flocculation (CBF) process to treat municipal wastewater. Chemically enhanced primary treatment (CEPT) process and primary sedimentation tank process were taken as the parallel-compared wastewater treatment processes. The experimental results show that under the same dosage, Zeta potential of the CBF process effluent is equal to that of the CEPT process, which indicates that flocculant in return sludge does not change the stabilization of particles in CBF reactor, and the biological flocculation is the key reason for CBF is superior to CEPT. In CBF process, good removal results are achieved for particles >10 microm and dissolved TOC with molecular weight >6 ku by chemical dosage, and biological flocculation can not only promote the removal of particles >10 microm and dissolved TOC with molecular weight >6 ku, but also have high capacity to remove small particles and dissolved TOC with small molecular weight, with the results that particles >3 tpm are removed completely and TOC with molecular weight of 2-6 ku are removed by 42.5% .

Mistletoe alkali inhibits peroxidation in rat liver and kidney.

AIM: To explore the antioxidant and free radical scavenger properties of mistletoe alkali (MA). METHODS: The antioxidant effect of mistletoe alkali on the oxidative stress induced by carbon tetrachloride (CCl4) in rats was investigated. The rats were divided into four groups (n = 8): CCl4-treated group (1 mL/kg body weight), MA -treated group (90 mg/kg), CCl4+MA-treated group and normal control group. After 4 wk of treatment, the level of malondialdehyde (MDA), a lipid peroxidation product (LPO) was measured in serum and homogenates of liver and kidney. Also, the level of glutathione (GSH), and activities of glutathione reductase (GR), glutathione peroxidase (GSPx), superoxide dismutase (SOD), and glutathione-S-transferase (GST) in liver and kidney were determined. Scavenging effects on hydroxyl free radicals produced in vitro by Fenton reaction were studied by ESR methods using 5,5-dimethyl-1-pyrroline-N-oxide (DMPO) as a spin trap reagent and H2O2/UV as the OH. source. Urinary 8-hydroxydeoxyguanosine (8-OHdG) was determined by competitive ELISA. RESULTS: In CCl4-treated group, the level of LPO in serum of liver and kidney was significantly increased compared to controls. The levels of GSH and enzyme activities of SOD, GSPx and GR in liver and kidney were significantly decreased in comparison with controls. In CCl4+MA-treated group, the changes in the levels of LPO in serum of liver and kidney were not statistically significant compared to controls. The levels of SOD, GSPx and GR in liver and kidney were significantly increased in comparison with controls. There was a significant difference in urinary excretion of 8-OHdG between the CCl4-treated and MA-treated groups. CONCLUSION: Oxidative stress may be a major mechanism for the toxicity of CCl4. MA has a protective effect against CCl4 toxicity by inhibiting the oxidative damage and stimulating GST activities. Thus, clinical application of MA should be considered in cases with carbon tetrachloride-induced injury.