Enhancing biomass and lipid accumulation by a novel microalga for unsterilized piggery biogas slurry remediation.

The cost and efficiency of an algal-BS treatment system are determined by the specific microalgal species and BS pretreatment method. This study examines the growth of a novel algae Chlorella sp. YSD-2 and the removal of nutrients from the BS using different pretreatment methods, including dilution ratio and sterilization. The highest biomass production (1.84 g L-1) was achieved in the 1:2 unsterilized biogas slurry, which was 2.03 times higher than that in the sterilized group, as well as higher lipid productivity (17.29 mg L-1 d-1). Nevertheless, the sterilized biogas slurry at a 1:1 dilution ratio exhibited the most notable nutrient-removal efficiency, with COD at 71.97%, TP at 91.32%, and TN at 88.80%. Additionally, the analysis of 16S rRNA sequencing revealed a significant alteration in the indigenous bacterial composition of the biogas slurry by microalgal treatment, with Proteobacteria and Cyanobacteria emerging as the predominant phyla, and unidentified_Cyanobacteria as the primary genus. These findings suggest that Chlorella sp. YSD-2 exhibits favorable tolerance and nutrient-removal capabilities in unsterilized, high-strength biogas slurry, along with high productivity of biomass and lipids. Consequently, these results offer a theoretical foundation for the development of an efficient and economically viable treatment method for algal-BS.

Study on the treatment of corn alcohol wastewater by the internal circulation anaerobic reactor.

To comprehensively assess the efficacy of employing the internal circulation (IC) anaerobic reactor for corn alcohol wastewater treatment and investigate its feasibility, this study focused on anaerobic digestion parameters, energy balance, and the composition of the prokaryotic microbial community. During the operation of the reactor, the hydraulic retention time was progressively reduced from 4.8 to 1.6 days while achieving an average organic loading rate of 12.46 kg chemical oxygen demand (COD)/(m3·d). Moreover, the removal rate of COD exceeded 98%, and the energy balance (ΔE) reached 10.29 kJ/g fed COD. The initial manifestation of organic acidosis in the reactor was a decline in gas production, which is primarily caused by propionic acid accumulation. The subsequent analysis revealed a high diversity of prokaryotes in granular sludge, with the predominant archaea primarily involved in methane production through the acetic acid pathway. The IC anaerobic reactor shows exceptional performance in treating corn alcohol wastewater by optimizing its operating conditions. Energy balance analysis confirmed the feasibility of the process. The findings of this study may offer valuable insights for optimizing control strategies and engineering applications.

Non-clinical pharmacokinetics study in rat plasma, tissues and excreta of honokiol derivative HM475 by UPLC-Q-TOF-MS/MS.

To provide the basis for further development and research of drugs, non-clinical pharmacokinetics studies were conducted on HM475, which is composed of natural active molecules honokiol and metformin through cyclization. In this paper, HM475 was studied from six aspects by gavage and intraperitoneal injection: 1) Acute toxicity of HM475 in rats, 2) Pharmacokinetic characteristics of HM475 in rats, 3) Distribution characteristics of HM475 in heart, liver, spleen, lung, and kidney, small intestine, fat and brain of rats, 4) Main metabolic pathways of HM475 in rats, 5) Excretion of HM475 in rats, 6) Determination of protein binding rate of HM475 in bovine plasma, rabbit plasma, and rat plasma. Acute toxicity of HM475 on SD rats was evaluated by maximum dose method. The metabolic analysis method of HM475 in rats was first established by UPLC-Q-TOF-MS/MS technology, and the pharmacokinetic characteristics of oral administration and intraperitoneal injection were studied. Experimental results showed that HM475 had no obvious acute toxicity. The absolute oral bioavailability of HM475 was 38.45 %, and the drug concentration in plasma was higher than that in tissues. Combined with the process characteristics of HM475 in vivo, it is inferred that HM475 has enterohepatic circulation. In this study, non-clinical pharmacokinetics were systematically studied to provide data support for the clinical pharmacokinetics and pharmacodynamics of HM475, to more accurately predict the pharmacokinetic behavior of HM475 in human body and provide scientific data for the compound to enter clinical research.

Isolation and Identification of Volatile Substances with Attractive Effects on Wohlfahrtia magnifica from Vagina of Bactrian Camel.

Vaginal myiasis is one of the most serious parasitic diseases in Bactrian camels. At present, there are no reports on biological control measures of the disease. In this paper, the metabolomic analysis of vaginal secretions from susceptible and non-susceptible camels was performed by ACQUITY UPLC H-Class Ultra Performance Liquid Chromatograph. The results matched in 140 vaginal compounds. Methylheptenone, 1-octen-3-ol, and propyl butyrate and their mixtures were selected for gas chromatography-electroantennography (GC-EAD), electroantennography (EAG), behavioral experiments and trapping experiments of Wohlfahrtia magnifica (W. magnifica). Results showed that the W. magnifica had EAG responses to the three compounds, respectively. The EAG responses of female flies to different concentrations of methylheptenone were significantly different, but to the others had no significant difference, and there was no significant difference in the same compounds between the different sexes. Behavioral and trapping experiments showed that methylheptenone and 1-octen-3-ol have significant attraction to W. magnifica, but there was no significant difference to propyl butyrate. When methylheptenone and 1-octen-3-ol were mixed in different proportions, it was found that a mixture at the ratio of 1:1 and 0.5:1 had extremely significant and significant attraction, respectively, to both male and female W. magnifica. The study showed that, except for propyl butyrate, the higher the concentrations of the other two compounds, the stronger the attractivity to the W. magnifica, and a mixture at the ratio of 1:1 could enhance the attractivity to the W. magnifica.

Biogas slurry application alters soil properties, reshapes the soil microbial community, and alleviates root rot of Panax notoginseng.

Background: Panax notoginseng is an important herbal medicine in China, where this crop is cultivated by replanting of seedlings. Root rot disease threatens the sustainability of P. notoginseng cultivation. Water flooding (WF) is widely used to control numerous soilborne diseases, and biogas slurry shows positive effects on the soil physiochemical properties and microbial community structure and has the potential to suppress soilborne pathogens. Hence, biogas slurry flooding (BSF) may be an effective approach for alleviating root rot disease of P. notoginseng; however, the underlying mechanism needs to be elucidated. Methods: In this study, we conducted a microcosm experiment to determine if BSF can reduce the abundance of pathogens in soil and, alleviate root rot of P. notoginseng. Microcosms, containing soil collected from a patch of P. notoginseng showing symptoms of root rot disease, were subjected to WF or BSF at two concentrations for two durations (15 and 30 days), after which the changes in their physicochemical properties were investigated. Culturable microorganisms and the root rot ratio were also estimated. We next compared changes in the microbial community structure of soils under BSF with changes in WF and untreated soils through high-throughput sequencing of bacterial 16S rRNA (16S) and fungal internal transcribed spacer (ITS) genes amplicon. Results: WF treatment did not obviously change the soil microbiota. In contrast, BSF treatment significantly altered the physicochemical properties and reshaped the bacterial and fungal communities, reduced the relative abundance of potential fungal pathogens (Fusarium, Cylindrocarpon, Alternaria, and Phoma), and suppressed culturable fungi and Fusarium. The changes in the microbial community structure corresponded to decreased root rot ratios. The mechanisms of fungal pathogen suppression by BSF involved several factors, including inducing anaerobic/conductive conditions, altering the soil physicochemical properties, enriching the anaerobic and culturable bacteria, and increasing the phylogenetic relatedness of the bacterial community. Conclusions: BSF application can reshape the soil microbial community, reduce the abundance of potential pathogens, and alleviate root rot in P. notoginseng. Thus, it is a promising practice for controlling root rot disease in P. notoginseng.

High nitrite-nitrogen stress intensity drives nitrite anaerobic oxidation to nitrate and inhibits methanogenesis.

Nitrite is an important intermediate in nitrogen metabolism. We explored the effect of nitrite-nitrogen stress intensity (NNSI) on nitrite metabolism and methanogenesis in anaerobic digestion. The results showed that the NNSI regulated microbial diversity, composition, and functions, and microbial community assembly was primarily shaped by stochastic processes. Moreover, the NNSI was negatively correlated with α-diversity and positively correlated with non-metric multi-dimensional scaling distance. Denitrification gradually increased with increasing NNSI; however, methanogenesis was gradually inhibited, which was primarily due to the inhibition of the aceticlastic methanogenesis pathway (i.e., Methanosaeta) and methylotrophic methanogenesis pathway (i.e., Candidatus_Methanofastidiosum). High NNSI (1882 ± 98.99 mg/L NO2--N) promoted nitrite anaerobic oxidation to nitrate and was favorable for dissimilatory nitrate reduction to ammonia (DNRA). We present evidence for the microbial transformation of nitrite under anaerobic conditions, with potential geochemical and evolutionary importance. As nitrogen oxides were already present on early Earth, our finding presents the possibility of a nitrogen cycle before the evolution of oxygenic photosynthesis.

Enhanced anaerobic digestion of swine manure via a coupled microbial electrolysis cell.

Microbial electrolysis cell coupled anaerobic digestion (MEC-AD) is a new technology in energy recovery and waste treatment, which could be used to recycle swine manure. Here, different applied voltage effects were studied using MEC-AD with swine manure as a substrate. The maximum cumulative biogas and methane yields, both occurring with 0.9 V, were 547.3 mL/g total solid (TS) and 347.7 mL/g TS, respectively. The increased energy can counterbalance the electrical input. First order, logistic, gompertz, and back-propagation artificial neural network (BP-ANN) models were used to study cumulative biogas and methane yields. The BP-ANN model was superior to the other three models. The maximum degradation rate of hemicellulose, cellulose, and lignin was 60.97%, 48.59%, and 31.59% at 0.9 V, respectively. The BP-ANN model establishes a model for cumulative biogas and methane yields using MEC-AD. Thus, MEC-AD enhanced biogas and methane production and accelerated substrate degradation at a suitable voltage.

Long noncoding RNA LIFR-AS1 suppresses proliferation, migration and invasion and promotes apoptosis through modulating miR-4262/NF-κB pathway in glioma.

AIM: This study aimed to explore the role of lncRNA leukemia inhibitory factor receptor antisense RNA 1 (LIFR-AS1) on glioma and its underlying molecular mechanism. METHODS: The expression of LIFR-AS1 and miR-4262 was detected by quantitative real-time polymerase chain reaction (qRT-RCR) in both glioma tissues and cell lines. Colony formation assay, 5-ethynyl-20-deoxyuridine (EdU) assay, flow cytometry and transwell assay were respectively conducted to detect cell clones, proliferation, apoptosis, migration and invasion. The effect of LIFR-AS1 on the chemoresistance to temozolomide (TMZ) of glioma cells was also analyzed. In addition, dual-luciferase reporter gene assay was performed to evaluate the luciferase activity. The expressions of nuclear factor-κB (NF-κB) p65, p-NF-κB p65 and inhibitor of κBα (IκBα) in glioma cells were measured by western blot. RESULTS: LIFR-AS1 was lowly expressed and miR-4262 was highly expressed in glioma tissues and cell lines. LIFR-AS1 overexpression inhibited the proliferation, migration and invasion and promoted apoptosis of glioma cells. LIFR-AS1 overexpression also reduced the chemoresistance to TMZ of glioma cells. Moreover, LIFR-AS1 overexpression suppressed the activation of NF-κB signaling pathway in glioma cells. miR-4262 was the target gene of LIFR-AS1. We also found that miR-4262 abrogated the functions of LIFR-AS1 on cell proliferation, apoptosis, migration and invasion of glioma cells in the NF-κB pathway. CONCLUSION: LIFR-AS1 could suppress the proliferation, migration and invasion and promote the apoptosis through modulating miR-4262/NF-κB pathway in glioma.

Comparison of long-term energy efficiency and microbial community dynamics of different reactors in response to increased loadings of water hyacinth juice.

Water hyacinth is considered to be among the worst invasive weed species globally, causing detrimental environmental and social problems worldwide. It rapidly grows, and therefore has significant potential as a resource. Due to its high moisture content (approximately 95%), the by-product obtained by dehydrating water hyacinth yields a considerable amount of water hyacinth juice (WHJ). In this study, we performed a comparative assessment of long-term energy efficiency, maximum treatment capacity limits, and microbial community dynamics of modified internal circulation (MIC) and up-flow anaerobic sludge blanket (UASB) reactors in response to increasing loadings of WHJ. The MIC reactor exhibited a higher energy recovery rate and stronger performance compared with the UASB reactor. The optimal organic loading rates of the MIC and UASB reactors were 17.93 and 8.85 kg chemical oxygen demand (COD)/m3/d, with methane conversion rates of 0.21 and 0.15 m3 CH4/kg COD, respectively. Furthermore, the engineering costs and project floor space required by the MIC reactor are less than those in the case of the UASB reactor. The high-throughput sequencing analysis indicated that the dominant phyla (e.g. Firmicutes and Bacteroidetes) were more abundant using the MIC reactor than with the UASB reactor, which may indicate WHJ degradation efficiency. Both reactors had similar predominant methanogens, suggesting that acetoclastic methanogenesis was the predominant metabolic pathway of methane formation. The results of this study provide new insights into the sustainable management of water hyacinth as a resource by establishing a regional ecosystem with biogas engineering applications.

Novel start-up process for the efficient degradation of high COD wastewater with up-flow anaerobic sludge blanket technology and a modified internal circulation reactor.

To avoid wastage of water resources and operating cost increases caused by the traditional start-up process of large amounts of dilution influent chemical oxygen demand (COD), a novel start-up process (NSP) was developed and verified with water hyacinth juice (WHJ) on an up-flow anaerobic sludge blanket (UASB) and modified internal circulation (MIC) reactor. Results show that UASB and MIC reactors were started successfully and that the MIC reactor exhibited a superior performance. The NSP time of the MIC reactor (46 days) was less than that of the UASB reactor (52 days), although the start-up organic loading rate (OLR) of the MIC reactor was higher than that of the UASB reactor. Interestingly, high-throughput sequencing analysis indicated that the reactor configuration significantly impacted the microbial diversity, however, the UASB and MIC reactors had similar predominant methanogens: Methanosaeta and Methanosarcina. Therefore, acetoclastic methanogenesis is the primary pathway of methane formation during WHJ treatment.

TGF-ß1 mediates lncRNA GAPLINC expression to promote the migration and invasion of non-small cell lung cancer.

Purpose: The present study aims to investigate the involvement of lncRNA GAPLINC in non-small lung cancer (NSCLC). Patients and methods: The study included 70 patients with NSCLC (39 males and 31 females, 33 to 68 years, 49.3 ± 6.4 years). RT-qPCR, transient cell transfections, measurement of in vitro cell migration and invasion abilities and western blot were carrying out during the research. Results: We showed that GAPLINC was up-regulated in NSCLC tissues and positively correlated with TGF-ß1. In vitro cell experiment showed that over-expression of TGF-ß1 significantly up-regulated the expression of GAPLINC, while over-expression of GAPLINC failed to affect TGF-ß1. Follow-up study showed that high GAPLINC level in NSCLC tissue was closely correlated with poor survival rate of NSCLC patients. Over-expressions of TGF-ß1 and GAPLINC resulted to accelerated migration and invasion of NSCLC cells. In addition, the silencing of GAPLINC siRNA attenuated the effect of TGF-ß1 treatment. Conclusion: TGF-ß1 may mediate lncRNA GAPLINC expression to promote NSCLC cell invasion and migration.

Effects of liver function, insulin resistance and inflammatory factors on vascular endothelial dilation function and prognosis of coronary heart disease patients complicated with NAFLD.

Effects of liver function, insulin resistance (IR) and inflammatory factors on vascular endothelial dilation function and prognosis of coronary heart disease (CHD) patients complicated with non-alcoholic fatty liver disease (NAFLD) were investigated. A total of 80 patients with CHD treated in Jinan Central Hospital from October 2016 to July 2017 were randomly enrolled and divided into the NAFLD group (n=41) and the simple CHD group (n=39). The IR, the vascular endothelial dilation function and the related inflammatory factors were also detected, followed by statistical analysis. The level of high-sensitivity C-reactive protein (hs-CRP), serum total bilirubin and alanine aminotransferase (ALT) levels and body mass index in the NAFLD group was decreased and the expression of tumor necrosis factor-α was increased compared with that in the simple CHD group (P<0.05). There was a linearly positive correlation between ALT and brachial artery diameter in the NAFLD group (r=0.311, P<0.05). There was a correlation between homeostasis model assessment-IR (HOMA-IR) and HOMA-ß indexes and brachial artery diameter, hs-CRP and brachial artery diameter in both groups (r=-0.128, r=0.219, P<0.05). The HOMA indexes in the NAFLD group were increased compared with those in the simple CHD group (P<0.01). There were significant differences in the intima-media thickness, number of carotid plaques and detection rate of carotid plaques (P<0.05). The risk of cardiovascular events within 10 years in the NAFLD group was increased compared with that in the simple CHD group. The differences of incidence of cardiovascular diseases (CVD)s were statistically significant (P<0.05). Therefore, The changes in liver function indexes, IR and related inflammatory factors in CHD patients complicated with NAFLD significantly affect the vascular endothelial dilation function, which also have some effects on the occurrence of CVDs.

Insulin resistance, blood glucose and inflammatory cytokine levels are risk factors for cardiovascular events in diabetic patients complicated with coronary heart disease.

This study was designed to investigate the relation of insulin resistance, blood glucose and inflammatory cytokines with cardiovascular events in diabetic patients complicated with coronary heart disease (CHD). A total of 120 patients with diabetes mellitus type 2 (T2DM) complicated with CHD admitted to Jinan Central Hospital from January 2015 to March 2016 were enrolled in this study. There were 60 cases complicated with cardiovascular events and 60 had no history of cardiovascular events; there were 40 cases with abnormal blood glucose, 40 with insulin resistance and 40 with elevated inflammatory cytokines. Changes in the levels of blood glucose, fasting serum insulin and inflammatory cytokines as well as changes in the homeostasis model assessment of insulin resistance indexes (HOMA-IR) were recorded and compared among groups of patients. Besides, changes in the Global Registry of Acute Coronary Events (GRACE) risk score and the incidence rate of cardiovascular events were also detected and multivariate logistic regression analysis was conducted so as to identify relevant risk factors. Our results showed the fasting blood glucose and the 2 h postprandial blood glucose levels in the non-cardiovascular event group were lower than those in the cardiovascular event group (P<0.05). However, levels of fasting serum insulin and HOMA-IR in the non-cardiovascular event group were significantly higher than those in the cardiovascular event group (P<0.05). Additionally, levels of tumor necrosis factor-α (TNF-α), interleukin-6 and C-reactive protein in the non-cardiovascular event group were significantly lower than those in the cardiovascular event group (P<0.05). Moreover, GRACE risk scores in patients with elevated inflammatory cytokines were higher than those in patients with insulin resistance and those in patients with blood glucose abnormalities (P<0.05). The incidence rate of cardiovascular events in patients with blood glucose abnormalities was lower than that in patients with insulin resistance and that in patients with elevated inflammatory cytokines. There was a positive correlation between TNF-α and HOMA-IR (P<0.05), and between HOMA-IR and the GRACE risk score (P<0.05). Blood glucose abnormalities, insulin resistance and inflammatory cytokines were all independent risk factors for cardiovascular events. Based on our findings, stronger inflammatory responses in patients with T2DM complicated with CHD lead to higher incidence rates of cardiovascular events. Besides that, elevated blood glucose and insulin resistance levels are also independent risk factors for cardiovascular events.

Optimization and kinetic study of methyl laurate synthesis using ionic liquid [Hnmp]HSO4 as a catalyst.

Methyl laurate was synthesized from lauric acid (LA) and methanol via an esterification reaction using ionic liquids (ILs) as catalysts. The efficiencies of three different catalysts, 1-methylimidazole hydrogen sulfate ([Hmim]HSO4), 1-methyl-2-pyrrolidonium hydrogen sulfate ([Hnmp]HSO4) and H2SO4, were compared. The effect of the methanol/LA molar ratio, reaction temperature, reaction time and catalyst dosage on the esterification rate of LA was investigated by single-factor experiments. Based on the single-factor experiments, the esterification of LA and methanol was optimized using response surface methodology. The results showed that the most effective catalyst was the IL [Hnmp]HSO4. The optimal conditions were as follows: [Hnmp]HSO4 dosage of 5.23%, methanol/LA molar ratio of 7.68 : 1, reaction time of 2.27 h and reaction temperature of 70°C. Under these conditions, the LA conversion of the esterification reached 98.58%. A kinetic study indicated that the esterification was a second-order reaction with an activation energy and a frequency factor of 68.45 kJ mol-1 and 1.9189 × 109 min-1, respectively. The catalytic activity of [Hnmp]HSO4 remained high after five cycles.

Metformin affects thyroid function in male rats.

An intriguing area of research in type 2 diabetes recently discovered association of metformin therapy with thyroid functional and morphological changes. We aimed to evaluate the external symptoms and biochemical indicators concerning thyroid function in rats treated with metformin. Male wistar rats were randomly divided into four groups: Group (D-/M-), Group (D-/M+), Group (D+/M-), and Group (D+/M+), according to whether they were induced to diabetic model or placed on metformin. Characteristics of food intake, body weight, and other external symptoms were recorded. Thyroid function, concluding serum thyrotropin (TSH), free triiodothyronine (FT3), free thyroxine (FT4), were measured. We found a significantly higher TSH and lower FT4 in rats in Group (D+/M-), compared with rats in Group (D-/M-), but no significant change in FT3 level. Rats on metformin treatment exhibited relatively lower body weight and symptoms like irritability and diarrhea, concomitant with marked increase in FT3 and FT4 , no matter if they were induced to diabetic model or not . A slight but significant reduction in TSH concentration was also observed in rats received metformin. These data reveal that metformin can modify thyroid function with corresponding clinical symptoms of hyperthyroidism in male rats. Metformin's contribution to suppress TSH and increase FT3, FT4 should arise our attention to its treatment interference in clinical practice.

Effects of shRNA-Mediated SOX9 Inhibition on Cell Proliferation and Apoptosis in Human HCC Cell Line Hep3B Mediated by Ultrasound-Targeted Microbubble Destruction (UTMD).

Hepatocellular carcinoma (HCC) is one of the most aggressive tumors in humans. The survival rate of patients is still very poor as current therapies offer limited treatment efficacy. Therefore, it is necessary to explore novel and more effective strategies to treat HCC. Recently, Ultrasound-targeted microbubble destruction (UTMD) has been shown to be a better alternative to viral vectors in delivering plasmid DNA into cells. In this study, we thus first determined the effect of combining UTMD with effectene on the transfection efficiency in human Hep3B cells. Transfection rate of the [effectene + shRNA-SOX9 + UTMD] group was the highest among the five groups, and were significantly higher than that of the [effectene + shRNA-SOX9] or [shRNA-SOX9 + UTMD] groups, while there was no significant difference between [shRNA-SOX9 alone] and [shRNA-SOX9 + UTMD] groups. Expression of SOX9 mRNA and protein was the lowest in effectene + shRNA-SOX9 + UTMD group. Moreover, transfection of shRNA-SOX9 with UTMD and effectene in combination could markedly inhibit the proliferation and induced cell apoptosis of Hep3B cells. These results suggest that the efficiency of gene delivery is remarkably increased when UTMD is combined with other transfection strategies, such as effectene. In conclusion, our research demonstrates that combining conventional transfection methods with UTMD achieves better transfection efficiency and that this can provide an improved gene delivery system for gene therapy.

[Screening on antiosteoporotic active parts of dipsacus radix based on zebrafish model].

OBJECTIVE: Prednisolone-induced osteoporosis model using zebrafish was used to screen the antiosteoporotic active parts of Dipsacus Radix, in order to investigate the applicability and rationality of the zebrafish model of osteoporosis. METHODS: Zebrafish larvae at 5 days post fertilization (dpf) were exposed with 25 micromol/L prednisolone and 0.5% DMSO for 48 h, then except one group of 25 micromol/L prednisolone, other groups of 25 micromol/L prednisolone were treated with a range of concentration (0.025, 0.25, 2.5, 25 microg crude drug/mL) of extract of Dipsacus Radix and its different concentration ethanol elution parts of macroporous resin with 25 micromol/L prednisolone. All groups were incubated in 24-well plates (28.5 degrees C) until 10 dpf. Zebrafish skeleton at 10 dpf were anesthetized and fixed for staining with alizarin red. Quantitative analysis of the stained area was performed by microscopic inspection and digital imaging methods to reflect the amount of zebrafish head skeleton mineralization. RESULTS: The results indicated that head skeleton mineral area and integrated optical density (IOD) of 25 micromol/L prednisolone model group were significantly decreased when compared with vehicle control group, and the extract of Dipsacus Radix and its 30%, 50%, 70% and 90% ethanol elution parts of macroporous resin rescued the further bone loss of zebrafish induced by prednisolone when compared with the model group. HPLC analysis indicated that components of 30%, 50%, 70% and 90% ethanol elution parts of macroporous resin containing saponins and nonsaponins components. CONCLUSION: Both saponins and nonsaponins can prevent bone loss of zebrafish induced by prednisolone. This novel osteoporosis zebrafish model was successfully used to screen antiosteoporotic active parts of Dipsacus Radix, which had advantages of simple, high efficiency and easy to perform.

[Establishment of zebrafish osteopenia model induced by dexamethasone].

Zebrafish was selected as model animal, and glucocorticoid dexamethasone was used as a model compound to establish a rapid and high efficient osteopenia model. Zebrafish larvae at 4 days post fertilization (dpf) were exposed to a serial concentrations of dexamethasone solutions, and 0.5% DMSO was selected as the vehicle control group. All groups were incubated in 24-well plates (28.5 degrees C) until 9 dpf. In addition, effects of 10 micromol x L(-1) dexamethasone on preventing against osteopenia induced by etidronate disodium were also investigated. Zebrafish bones at 9 dpf were stained with alizarin red. Quantitative analysis of the stained area was performed by microscopic inspection and digital imaging methods to reflect the amount of bone mineralization. Results showed that dexamethasone group at 2.5, 10 and 25 micromol x L(-1) can decrease the staining area and the staining optical density values of zebrafish head bones when compared with the vehicle control group (0.5% DMSO), which suggested that dexamethasone can significantly reduce the zebrafish mineralized bone and the bone mineral density. Results also showed that 15 and 30 microg x mL(-1) etidronate disodium can increase the mineralized matrix of zebrafish head bone and prevent against osteopenia induced by dexamethasone. In conclusion, the study indicated that zebrafish can be an idea osteopenia model induced by dexamethasone.

[Metabolism study of asperosaponin VI by using zebrafish].

Model organism zebrafish was used to study metabolism of asperosaponin VI from Dipsacus asper Wall. ex Henry for the first time. Metabolic components of asperosaponin VI after exposing to zebrafish for 24 h were identified by high performance liquid chromatography--electrospray mass spectrometry (HPLC-ESI-MS), the separation was performed with a Zorbax C18 column using a binary gradient elution of 0.05% formic acetonitrile--0.05% formic acid water. The quasi-molecular ions of compounds in both negative and positive mode were observed for molecule mass information, and the potential structures were identified by attentive study on the deglycosylation metabolites and one hydroxylation metabolite of asperosaponin VI. The results were highly in consistent with metabolism of asperosaponin VI in rat. It can be concluded that zebrafish model can wonderfully imitate current metabolic model with advantages of small amount of lower cost, far less amount compound, higher efficiency and more simple, and can reflect integrated metabolism results of in vivo method. Zebrafish metabolic model may become a novel organism model for quick predication on metabolism of even mircoamount compound, which can enrich the available models greatly.

Metabolism of tanshinone IIA, cryptotanshinone and tanshinone I from Radix Salvia miltiorrhiza in zebrafish.

The study aimed to investigate the potential of zebrafish in imitating mammal phase I metabolism of natural compounds. Three diterpenoid quinones from Radix Salvia miltiorrhiza, namely tanshinone IIA (TIIA), cryptotanshinone (Cry) and tanshinone I (TI) were selected as model compounds, and their metabolites mediated by zebrafish were characterized using a high-performance liquid chromatography coupled ion-trap mass spectrometry (HPLC/IT-MSn) method with electrospray ionization in positive mode. The separation was performed with a Zorbax C-18 column using a binary gradient elution of 0.05% formic acid acetonitrile/0.05% formic acid water. According to the MS spectra and after comparison with reference standards and literature reports, hydroxylation, dehydrogenation or D-ring hydrolysis metabolites of TIIA and Cry but not of TI were characterized, which coincided with those reported using regular in vivo or in vitro metabolic analysis methods, thus verifying that zebrafish can successfully imitate mammalian phase I metabolism which instills further confidence in using zebrafish as a novel and prospective metabolism model.