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Type 2 diabetes mellitus (T2DM), a chronic low-grade inflammatory disease with high morbidity and mortality, is a serious threat to public health. Previously we demonstrated that a traditional Chinese medicine formulation, Jiedu Tongluo Tiaogan Formula (JDTL), exerted a favorable hypoglycemic effect due to unknown molecular mechanisms involving interactions among JDTL compounds and various cellular components. This study aimed to explore JDTL mechanisms for alleviating hyperglycemia using an integrated strategy incorporating system pharmacology, bioinformatics analysis, and experimental verification. This strategy entailed initial elucidation of JDTL chemical composition using fingerprint analysis via high performance liquid chromatography (HPLC). Next, functions of putative shared target genes and associated pathways were deduced using GO and KEGG pathway enrichment and molecular docking analyses. Ultimately, targets associated with JTDL anti-T2DM effects were found to be functionally associated with biological functions related to lipopolysaccharide and cytokine receptor binding. These results implicated PI3K-Akt signaling pathway involvement in JDTL anti-T2DM effects, as this pathway had been previously shown to play significant roles in glucose and lipid metabolism-related diseases. Furthermore, addition of JDTL to INS-1 and HepG2 cell cultures stimulated cellular mRNA-level and protein-level expression leading to enhanced production of IRS1, Akt, and PI3K. In summary, here JDTL bioactive ingredients, potential targets, and molecular mechanisms underlying JDTL anti-T2DM effects were identified using a multi-component, multi-target, and multi-channel analytical approach, thus providing an important scientific foundation to facilitate development of new drugs mechanistic strategies for preventing and treating T2DM.

The complex [Dy4(L1)2(CH3COO)4(DMF)3] (1) was obtained by using 2,6-bis(2-((E)-(8-hydroxyquinolin-2-yl)methylene)hydrazine-1-carbonyl)pyridine 1-oxide (H4L1) and Dy(CH3COO)3·4H2O under 100 in N,N-dimethylformamide (DMF) solvent. Structure analyzing showed that Dy(III) ions had two types of configuration with D2d and C4v. AC magnetic susceptibilities exhibited that complex 1 was an obvious single-molecule magnet (SMM) with obvious ac susceptibilities at zero Oe static magnetic field over the frequency range of 9–999 Hz. These findings demonstrated potential applications of polydentate ligands for the synthesis of SMMs.

We successfully obtained two type of heterometallic complexes [Fe4Dy2(L1)2(µ4-O)2(piv)4(H2O)2(NO3)4]·CH3CN·CH3OH (1) and [Fe6Dy2(L1)2(µ4-O)2(CH3COO)6(OH)2(H2O)2(NO3)6]·CH3CN (2) by using 2,2′-((2-((2-hydroxyethyl)amino)cyclohexyl)azanediyl)bis(ethan-1-ol) (H3L1), Dy(NO3)3·6H2O, FeCl3 and pivalic acid (piv) or acetic acid under room temperature in different solvent. Crystal structure showed that the central structure of [ $${{\text{Fe}}}_{4}^{{\text{III}}}$$ (µ4-O)2]8+ arranged in what was often termed an absent cubic shape, although here the Fe centers were strictly coplanar. Each Fe3 triangle was connected to the capping Dy(III) through µ4-O bridges for complex 1, and each Fe3 triangle was connected to the capping Fe(III) through µ4-O bridges for complex 2. It might be due to the steric effect that the coordination atoms of piv ligands chelated and blocked the structure. AC magnetic susceptibilities exhibited that complex 1 and 2 showed out-of-phase (χ″) component of ac-susceptibilities at zero Oe static magnetic field over the frequency range of 9–999 Hz, which energy barriers were 0.44 and 0.13 K. And their energy barriers were 1.2 and 12.2 K at applied 1000 Oe static magnetic field.

Food is people’s primal want. A reasonable diet and healthy food not only provide nutrients for human growth but also contribute to disease prevention and treatment, while following an unhealthy diet can lead to an increased risk of many diseases, especially metabolic disorders, such as diabetes. Nature is enriched with different food sources, and it seems that purely natural products are more in line with the current concept of health, which enhance the formation of the notion that “Food/Diet Supplements from Natural Sources as a Medicine.” As a delicious fruit, the medicinal values such as anticancer, antibacterial, antioxidation, and antiglycating properties of lychee have been found. Lychee (Litchi in Chinese) is a subtropical fruit plant belonging to the family Sapindaceae. It has been widely cultivated in warm climates worldwide, particularly in China, for thousands of years. In recent years, various phytochemical components such as quercetin, procyanidin A2, and (2R)-naringenin-7-O-(3-O-αL-rhamnopyranosyl-β-D-glucopyranoside) have been identified in a lychee seed, which may lend a lychee seed as a relatively safe and inexpensive adjuvant treatment for diabetes and diabetic complications. In fact, accumulating evidence has shown that lychee seed, lychee seed extracts, and related compounds have promising antihyperglycemic activities, including improving insulin resistance, anti-inflammatory effect, lipid regulation, neuroprotection, antineurotoxic effect, and renoprotection effect. In this review, we summarized publications on antiglycemic effects and mechanisms of lychee seed, lychee seed extracts, and related compounds, which included their efficacies as a cure for diabetes and diabetic complications in cells, animals, and humans, attempting to obtain a robust evidence basis for the clinical application and value of lychee seed.

Background: T2DM is considered to be a chronic low-grade inflammatory disease, because of its high morbidity and mortality, diabetes poses a tremendous potential threat to public health. Our previous studies have shown that a traditional Chinese medicine formula, Jiedu Tongluo Tiaogan Formula (JDTL) exerts favorable hypoglycemic effect, however, its molecular mechanism and the interaction among various components need to be further elucidated. This study aimed to explore the mechanism of JDTL in the treatment of T2DM using an integrated strategy of system pharmacology, bioinformatics analysis, and experimental verification. Materials and Methods: First, the compounds of JDTL were searched from public databases, and the "compound-target " network was constructed to predict the potential active components and targets. Subsequently, bioinformatics analysis was used to identify potential targets and signaling pathways, including PPI, GO pathways and KEGG pathways. Finally, the pharmacological effects and mechanisms of JDTL were verified by molecular docking and cell experiments. Results: Analysis by GO and KEGG pathway enrichment revealed that these targets were associated with lipopolysaccharide, membrane microdomain, cytokine receptor binding, and JDTL could regulate the PI3K/Akt signaling pathway. Furthermore, we have proved that JDTL could improve the mRNA expression and protein expression of IRS1, AKT, and PI3K in the INS-1 cell and HepG2 cells. Conclusion: The present study elucidated the active ingredients, potential targets, and molecular mechanism of JDTL in the treatment of T2DM, and revealed the characteristics of JDTL in multi-component, multi-target and multi-channel. It also provided an important scientific basis for new drug development and mechanism research of T2DM.

Recently, the bulk-heterojunction (BHJ) organic photovoltaic cells (OPVs) have attracted more attention owing to their potentials in low-weight, cost-effective, semi-transparent, colorful, and flexible. As one of the most promising candidates non-fullerene small molecule acceptors (SMA) in BHJ OPVs, fluoranthene-fused imide (FFI) derivatives can effectively regulate energy level and absorption by functional modification of active sites on its backbone. Nevertheless, it is still a challenge to design molecules based on FFI materials to achieve significant improvements in optoelectronic performance. Therefore, the key to this work is to explore the influence of introducing functional substituents at different active sites of FFIs on the photoelectric properties. By introducing different number of substituent groups with different electron-withdrawing and electron-donating capacities at FFI backbones active sites, could lead to construct “D-A-D” structures. By density functional theory (DFT) and time-dependent density functional theory (TD-DFT) calculations, we demonstrated that constructing “D-A-D” structure in FFI derivatives wouldn’t impact apparently on charge transfer process, because the nearly vertical dihedral angles exhibit in these geometrical configurations between between “D units” and “A units”. This has become a main factor limiting the performance improvement of FFI-based SMAs with “D-A-D” structures. On the contrary, it would be an effective approach to control the number of electron-withdrawing groups reasonably at FFI backbone in order to design more excellent performance for FFIs.

Background. Rhizoma coptidis (RC) showed a significant effect on PCOS, but its mechanism in PCOS remains unclear. Methods. The components of RC were searched by TCMSP. The Smiles number of the active ingredients was queried through PubChem, and the predicted targets were obtained from the SwissTargetPrediction database. The DrugBank, GeneCards, and DisGeNET databases were retrieved to acquire the related targets of PCOS. Then, the network of compound-target was constructed. The core targets were analyzed using protein-protein interaction (PPI) analysis, and the binding activities were verified by molecular docking. The enriched pathways of key targets were examined by GO and KEGG. Results. 13 components and 250 targets of RC on PCOS were screened. The core network was filtered based on topological parameters, and the key components were palmatine, berberine, berberrubine, quercetin, and epiberberine. The key targets included DRD2, SLC6A4, CDK2, DPP4, ESR1, AKT2, PGR, and AKT1. Molecular docking displayed that the active ingredients of RC had good binding activities with potential targets of PCOS. After enrichment analysis, 30 functional pathways were obtained, including neuroactive ligand-receptor interaction, dopaminergic synapse, and cAMP signaling pathway. Conclusion. In summary, this study clarified the potential effect of RC on PCOS, which is helpful to provide references for clinical practice. It is also conducive to the secondary development of RC and its monomer components.

Background：Insomnia is a major global public health issue with a high incidence, which presents a significant economic burden. Importantly, insomnia is often accompanied by a myriad of symptoms during the daytime, the most common being insomnia dizziness, headache, malaise, fatigue, anxiety, and even contribute to several diseases. However, the action mode of multi-component and multi-target for Chinese medicine could be a promising therapy for insomnia. According to the previous research, the ZaoRenDiHuang (ZRDH) Capsules showed the noteworthy anti-insomnia effect. Up to now, active ingredients, potential targets, and signaling pathways and mechanism of action are not yet clear. In this study, network pharmacology was employed to elucidate the potential anti-insomnia mechanism of ZRDH.Methods：In this study, an integrated pharmacology approach was implemented, which involved evaluation of absorption, distribution, metabolism and excretion of ZRDH, data mining of the insomnia targets, protein-protein interaction (PPI) network analysis, enrichment analysis, and molecular docking simulation, to predict the bioactive components, potential targets, and molecular mechanism of ZRDH for insomnia.Results：In this work, 44 anti-insomnia components of ZRDH and 65 anti-insomnia targets of insomnia were filtrated through database mining. The Drug-Disease network was constructed andfive key components Jujuboside A, Schizandrin A, Schizandrin C, Schizandrin B, and Spinosin, were further obtained. Sixty-five key targets were identified by topological analysis. Sequential studies turned out, NMURl, CAlCR, GABA, TAER2, ORDS, CYS1TR2, HTR1B, TLR4 were the common key targets. Docking studies indicated that the bioactive compounds could stably bind the pockets of target proteins. The findings of Gene Ontology (GO) terms and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway annotation suggested that the Neuroactive ligand−receptor interaction, Serotonergic synapse CAMP signaling pathway, HIF−1a signaling pathway, Toll−like receptor signaling pathway, anti-insomnia through data mining and network analysis.Conclusion: In sunmmary, potential mechanisms involved in ZRDH treatment for insomnia involves multiple components and multiple target points as well as multiple pathways. These findings may offer a profile for further investigations of the anti-fibrotic mechanism of ZRDH.

Background Diabetic retinopathy (DR) is the leading cause of blindness in many countries. The current treatment for non-proliferative DR (NPDR) using Western medicine (WM) alone is insufficient. At present, the combination of NPDR treatment with traditional Chinese medicine (TCM) and WM is universally applied. We aimed to evaluate the effectiveness and safety of TCM as an add-on for NPDR using a systematic review and meta-analysis. Method Data from randomized controlled trials (RCTs) of TCM for NPDR treatment along with WM before July 6, 2019, were collected from the China National Knowledge Infrastructure, Wanfang Database, China Biomedical Database, Pubmed, Embase, and Cochrane Library. Relevant data were extracted by two reviewers. I2 statistics was adopted to appraise heterogeneity. If I2 Result Eighteen RCTs (1522 patients) were included based on the inclusion and exclusion criteria. The results showed that compared with WM alone, TCM (including Compound Xueshuantong Capsule, Qiming Granule, and others) combined with WM for NPDR could improve the overall effiicacy [n = 1686, RR 1.24 (1.18,1.30), P I2 = 0%], and reduce the influence of risk factors related to NPDR, such as glycated hemoglobin level [n = 360, MD − 0.85 (− 1.28, − 0.41), P = 0.0001, I2 = 72%], triglyceride (P P = 0.0008). Moreover, no serious adverse events were reported. Conclusion Compared with WM alone, TCM + WM could significantly improve NPDR and also reduce the correlation levels of risk factors, such as hyperglycemia, dyslipidemia. However, the small sample included in the study might lead to a publication bias, and therefore, our results should be treated with caution.

Background Diabetic kidney disease (DKD) is the main cause of end-stage kidney disease and has become a heavy economic and social burden due to its high prevalence and morbidity. The most effective strategy is that patients with DKD should be diagnosed and treated early. Preliminary studies showed that the Chinese herbal Tangshen Formula (TSF) may delay the progression of DKD, reducing microalbuminuria and macroalbuminuria and improving renal function. We designed a randomized, double-blind, placebo-controlled trial to evaluate the efficacy of TSF in patients with DKD. Methods/design This trial is a 13-center, randomized, double-blind, placebo-controlled study. A total of 632 participants will be randomized in a 1:1 ratio to an experiment group (TSF plus losartan) and a control group (placebo plus losartan). The trial cycle will last 24 weeks. The primary outcome will be the change in the urine microalbumin–creatinine ratio from baseline to week 24. The secondary outcome will be the change in the rate of progression to the clinical proteinuria period after intervention, the rate of urine microalbumin negative conversion, the rate of normal urinary microalbumin, the doubling rate of the baseline creatinine value and the glomerular filtration rate between the two groups. Safety in medication will also be evaluated. Discussion We hypothesize that patients with type 2 diabetes in the early stage of DKD will benefit from TSF. If successful, this study will provide evidence-based recommendations for clinicians. Trial registration ClinicalTrials.gov, NCT03009864. Registered January 2017.

As an important means of regulating ligands, steric hindrance effect is always the first choice to construct structures. It is rare to assemble complexes with different topological and structural types by increasing or decreasing the steric hindrance of ligands and to regulate their properties. Here, we successfully obtained two types of lanthanide structures [Ln(H2L1)(NO3)3]n (1: Dy; 2: Gd, H2L1 ​= ​6,6'-((1E,1′E)-((2,2-dimethylpropane-1,3-diyl)bis(azanylylidene))bis(methanylylidene))bis(2-methoxyphenol)) and [Ln2(H2L2)3(NO3)6]·(CH3CN)x·(CH3OH)y (3: Dy, x ​= ​3, y ​= ​0; 4: Gd, x ​= ​y ​= ​2, H2L2 ​= ​6,6'-((1E,1′E)-((2,2-dimethylpropane-1,3-diyl)bis(azanylylidene))bis(methanylylidene))bis(2-ethoxyphenol)), which are controlled by ligands with the steric hindrance effect. For the all structures, the ligands are distorted when they coordinate with lanthanide ions, which make the ligands form helical structures around Ln(III) ions. For chains 1 and 2, there is a single twisted ligand bridging to form a one-dimensional spiral chain between the Ln(III) ion and the Ln(III) ion. But for complexes 3 and 4, there is a helical dinuclear structure formed by the cross-bridging of a double twisted ligand between the Ln(III) ion and the Ln(III) ion. AC magnetic susceptibilities show that chain 1 is a single-molecule magnet (SMM), which exhibits obviously frequency dependent behavior with the energy barriers of 39.6 (3) K and 44.6 (5) K, but complex 3 shows field-induced single-molecule magnetic behavior. The magnetocaloric effects (MCEs) show that chain 2 has bigger value of S than that of complex 4.

Perfluorinated compounds (PFCs) have received increasing attention worldwide recently because of potential risk to aquatic environment and living organisms. Herein, occurrence and spatial distributions of 17 selected PFCs were investigated in surface water adjacent to potential industrial emission categories in Shanghai. The results showed the distributions of PFCs in the ambient rivers were greatly affected by those industrial sources. Perfluorooctanoic acid (PFOA) and other short-chain PFCs such as perfluoropentanoic acid (PFPeA) and perfluorooctanesulfonate (PFBS) were detected as the predominant species in all samples. Specifically, the total concentrations of PFCs (∑PFCs) near the airport ranged from 142 to 264ngL-1, with PFOA, PFPeA, and PFBS as most prevalent. While near the fluorochemical plant and metal plating, concentrations of ∑PFCs ranged from 200 to 2143ngL-1 and 211ngL-1 to 705ngL-1; and PFOA was the predominant individual PFCs, with the highest concentration of 1985ngL-1. However, concentrations of PFOS were found at relatively low level, which ranged from < 0.06 to 4.44ngL-1. The Spearman correlation analysis of concentration of individual PFCs showed that PFOA and perfluorohexanoic acid (PFHxA) was positive, while the correlation between PFOA and perfluorohexansulfonate (PFHxS) was negative near the airport, indicating PFOA and PFHxA may share common sources. Preliminary ecological risk evaluation of PFCs in adjacent water of the industrial emission areas suggested these emission categories posed higher risks than other area, although the risk level was still relatively safe.

In the following paper, a Re(I) complex with electron-withdrawing oxadiazole group in its diamine ligand (denoted as N-N) was synthesized. Its single crystal structure analysis confirmed the successful synthesis of both ligand and complex. The central Re(I) ion localized in a traditional octahedral coordination environment. The diamine ligand 2-(pyridin-2-yl)-5-(p-tolyl)-1,3,4-oxadiazole (denoted as PPOZ) took a coplanar structure and the corresponding face-to-face π-π attraction between PPOZ ligands made the Re(I) complex molecules adjust a highly ordered arrangement which was positive to improve emissive performance. In order to repress the excited state geometric relaxation and further improve the emissive performance, the Re(I) complex was doped into a polymer host poly(vinylpyrrolidone) via electrospinning, resulting in composite fibers. The morphology of those composite fibers was analyzed by electron microscopy. The photophysical comparison between bulk sample and composite fibers indicated that the composite fibers showed emission blue shift, longer excited state lifetime and improved photostability. Further analysis suggested that the excited state geometric relaxation could be effectively repressed when the Re(I) complex molecules were immobilized in the polymer matrix, leading to above variations.

A new type of foam fractionation column with spiral internal had been designed for enhancing the foam drainage and thus for the removal of minute hazardous materials. Chromium was separated from wastewater using the continuous foam separation method. According to the similar physical behavior of foam separation and chemical reaction processes, the equivalent chemical reaction constant was introduced, Establishment concentrate distributed mathematical model of the whole tower under the condition of continuous foam separation with SDS as active agent. The model has been tested by experiments. The result indicated that foam separation process could be regarded as a first order reaction, and diffusion coefficient was 4.86cm2/s.

Background Our previous study showed an efficient targeting of islets of Langerhans by adenoviral injection via the celiac trunk. Unexpectedly, none of the endothelial cells was infected given the direct contact between adenoviruses and the capillary wall. The present study intended to provide an efficient approach for adenoviral targeting of the microcapillary endothelial cells in the pancreas. Methods We prepared microspheres of poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) (PHBHHx) with a size comparable to the diameter of capillary (5–10 µm). Scanning electron microscopy was applied to verify that adenoviruses carrying a green fluorescence protein gene were complexed with PHBHHx-microspheres after 30 min of co-incubation. The complexes were then injected into the pancreas of mice via the celiac trunk. Results Approximately 40% of endothelial cells in the pancreas were labeled 5 days after surgery. Islet cells were labeled occasionally, whereas labeling of the acinar and ductal tissues was barely detectable. Endothelium targeting was inefficient in other internal organs. Consistent with the reported superior tissue compatibility of PHBHHx, no discernable microspheres were found in all of the organs examined. Furthermore, splenocyte activation was dampened when adenoviruses were complexed with the microspheres. Conclusions The present study has established an approach for efficient pancreatic capillary targeting by using microsphere-adenoviral complexes. This procedure could be invaluable for the treatment of capillary-related diseases. Copyright © 2012 John Wiley & Sons, Ltd.

We wished to elucidate a potential role of the tumor necrosis factor-like weak inducer of apoptosis (TWEAK)/fibroblast growth factor-inducible molecule 14 (Fn14) axis in myocardial fibrosis. Stimulation of neonatal rat cardiac fibroblasts (CFs) with TWEAK could increase CFs numbers and collagen synthesis. Conversely, when CFs were pretreated with siRNA against Fn14, induction of cell proliferation and collagen synthesis by TWEAK were inhibited. Pretreatment with TWEAK on CFs induced activation of the nuclear factor-kappaB (NF-кB) pathway and subsequently increased the production of metalloproteinase-9 (MMP-9). Cell treatment with siRNA against Fn14 led to inhibition of the NF-кB pathway. Additionally, after stimulation of cell with ammonium pyrrolidine dithiocarbamate, cell proliferation and collagen synthesis induced by NF-кB and the upregulation of MMP-9 production were inhibited. The present study suggested that the TWEAK/Fn14 axis increased cell proliferation and collagen synthesis by activating the NF-кB pathway and increasing MMP-9 activity. This axis may be important for regulating myocardial fibrosis.

In order to enzymatically produce chitooligosaccharide using the crude enzyme preparation from Bacillus cereus D-11, we first studied the optimal reaction conditions. It was found that the optimal temperature for hydrolysis of chitosan was 55oC. The ratio of enzyme/substrate should not be lower than 0.13 U/mg in the reaction mixture. The enzyme activity was stable below 50oC. The products of enzymatic reaction were analyzed by both thin layer chromatography and high performance liquid chromatography. Under the appropriate condition, chitosan was hydrolyzed using the enzyme preparation. The resulting chitooligosaccharides were purified and separated by Dowex (H+) ion exchange chromatography. From 4 g soluble chitosan, 0.95 g (GlcN)2, 1.43 g (GlcN)3, and 1.18 g (GlcN)4 were recovered.

Cr(III)-containing wastewater was treated by continuous foam separation process. The factors affecting the Cr(III) removal efficiency were researched. The optimal operating conditions determined by orthogonal experiments are as follows:wastewater pH 6.50,gas flow 400ml/min,cationic surfactant dosage 50mg/L.When the mass concentration of Cr(III) in the influent is 7.5mg/L,the effluent can meet the national discharge standard with 98% of Cr(III) removal rate for continuous running. The kinetic experimental results show that the foam separation of Cr(III) accords with the first order kinetic equation. The results of continuous experiments in a scale-up foam separation column indicate which reach the Second Wastewater Discharge Standard of Country.

Lysobacter antibioticus HS124 isolated from pepper rhizosphere soil produced catechol type siderophore. Purified siderophore by Diaion HP-20 and silica gel column chromatography showed several hydroxyl functio nal groups adjacent to benzene rings by analysis of 1H NMR spectroscopy. The strain HS124 showed different activities to suppress Phytophthora capsici with different concentrations of exogenous Fe (III) in minimal medium where antifungal activity with 100 μM Fe (III) was approximately 1.5 times higher than in absence of Fe (III). Bacterial population in this Fe (III)-amended medium was also highest with 8.9×10 8 CFU ml -1 which also corresponded to the strongest siderophore activity. When grown in rich medium (minimal medium with N, P 2 O 5 , K 2 O and glucose), HS124 exhibited approximately 2 times stronger antifungal activity compared to minimal medium. In pot trials, treatments of bacterial culture grown in rich medium with (C1) or without (C2) 100 μM Fe (III) exhibited a high protection of pepper plants from disease, compared to medium only with (M1) or without (M2) 100 μM Fe (III). Especially, treatment C1 showed the best disease control effect of about 70 %. Thus, the strain HS124 should be recommended as a potential biocontrol agent against P. capsici in pepper.

One novel highly cross-linked, shape-controllable molecularly imprinted sol-gel film (MISF) with good thermal and chemical stability and three-dimensional network porous structure has been developed. The binding characteristic of the imprinted film to bifenthrin was evaluated by equilibrium binding experiments and imprinting-induced promotion of binding. The MISF was modified into the wells of 96 micro-well plate combined with chemiluminescence (CL) technology to establish the molecular imprinting-chemiluminescence (MI-CL) sensor. The high throughput and high selective detection sensor of bifenthrin was applied in vegetable and food samples with satisfactory results.

Streptococcus pneumoniae is a major cause of invasive infection in young infants and older adults. There are currently 90 capsular serotypes identified and 23 serotypes (1, 2, 3, 4, 5, 6B, 7F, 8, 9N, 9V, 10A, 11A, 12F, 14, 15B, 17F, 18C, 19F, 19A, 20, 22F, 23F, and 33F) are responsible for about 90% of invasive disease. Among the more than 90 different S. pneumoniae serotypes, serotype 19A is globally very prevalent. A simplified purification procedure including adjustment of cell lysate pH to 4.5, fractionation with 50- 80% ethanol, and dialysis rendered capsular polysaccharide (CPS) in a yield of 31.32 +/- 3.11 mg from 1 l culture (75% recovery after lyses). The product contained only 69.6 microng of protein (99.78% purity) and 0.8 mg (sum of the precipitants from 50~60%, 60~70%, and 70~80%) of nucleic acid (97.45% purity). The purified CPS was conjugated with bovine serum albumin; the product size ranged from 100 to 180 kDa.

A strain of phosphate solubilizing bacterium was isolated from rhizosphere and identified as Burkholderia sp. by 16S-rRNA gene sequence analyses. The bacterium was found to release gluconic acid and the solubilization of hydroxyapatite in the liquid medium by a significant drop in pH to 3.7 from an initial pH 7.0. The soluble-P concentration continuously increased during the incubation periods and the total amount of soluble P released in culture filtrate was detected at 990 mg L -1 after 10 days of inoculation. Most promoted maize growth was found in the standard NPK (240-120-120 kg ha -1 ) soil inoculation with Burkholderia sp. (Twenty milliliters/ plant, 106 CFU) and also in the absence of Burkholderia sp. inoculation, the soil amended with only 2/3 levels of P gave significant higher plant yield compared to 1/3 levels of P or without P supplementation.

The processing technique of bamboo particle board using laccase-treated calcium lignosulfonate was investigated and optimized. We studied the influence of enzyme amount, calcium ligosulfonate amount and hot pressing temperature on different particle board properties. All three factors were significant difference at different levels. There has significant differences between enzyme amount and Modulus of Rupture (MOR)，Modulus of Elasticity (MOE), Thickness Swelling (TS) (P

Hydroquinone galactoside (HQ-Gal) as a potential skin whitening agent was synthesized by the reaction of lactase (beta-galactosidase) from Kluyveromyces lactis, Aspergillus oryzae, Bacillus circulans, and Thermus sp. with lactose as a donor and HQ as an acceptor. Among these lactases, the acceptor reaction involving HQ and lactose with K. lactis lactase showed a higher conversion ratio to HQ-Gal (60.27%). HQ-Gal was purified using butanol partitioning and silica gel column chromatography. The structure of the purified HQ-Gal was determined by nuclear magnetic resonance, and the ionic product was observed at m/z 295 (C12H16O7Na)+ using matrix assisted laser desorption ionization time-of-flight mass spectrometry. HQ-Gal was identified as 4-hydroxyphenyl-beta-d-galactopyranoside. The optimum conditions for HQ-Gal synthesis by K. lactis determined using response surface methodology were 50 mM HQ, 60 mM lactose, and 20 U mL(-1) lactase. These conditions produced a yield of 2.01 g L(-1) HQ-Gal. The half maximal inhibitory concentration (IC50) of diphenylpicrylhydrazyl scavenging activity was 3.31 mM, indicating a similar antioxidant activity compared to beta-arbutin (IC50=3.95 mM). The Ki value of HQ-Gal (0.75 mM) against tyrosinase was smaller than that of beta-arbutin (Ki=1.97 mM), indicating its superiority as an inhibitor. HQ-Gal inhibited (23%) melanin synthesis without being significantly toxic to the cells, while beta-arbutin exhibited only 8% reduction of melanin synthesis in B16 melanoma cells compared with the control. These results indicate that HQ-Gal may be a suitable functional component in the cosmetics industry.

The ways of chemical component analysis, chemical analysis photoelectron energy spectrum (ESCA) analysis and X ray diffraction were used to research and analyze the variation law of material (wood) chemical component, functional groups and cellulose crystallinity in the process of making board. The results showed that：(1) The maximum contents of holocellulose and pentosan were wood, in the next place were fiberboard, and the minimum one were binderless fiberboard. The maximum contents of α-cellulose were woodfiber, in the next place were wood, and the minimum one were binderless fiberboard. The minimum contents of xylogen were wood, in the next place were woodfiber, and the maximum one were binderless fiberboard. (2) The C1and C3 functional groups relative contents of woodfiber and binderless fiberboard were higher than wood. The C2 functional groups relative contents of wood were higher than woodfiber and binderless fiberboard. The C4 functional groups relative contents of woodfiber were zero, and the binderless fiberboards were higher than wood. (3)The maximum cellulose relative crystallinity was binderless fiberboard, in the next place was fiberboard, and the minimum one was wood.

Grinding is an energy-intensive process in which the heat generated can cause various types of thermal damage to workpiece. Many theoretical, empirical or numerical models have been developed to predict grinding temperature. However, these models are not directly applicable for coated workpieces. Tools or other parts are coated with hard materials like tungsten carbide, ceramics or polycrystalline diamond to increase their surface hardness and prolong their life expectancy. In this paper, an empirical model is proposed to predict the maximum grinding temperature of coated workpieces. Experimental and numerical studies are carried out to validate the model. The results indicated that the new model is able to accurately predict grinding temperature.

In adult mammalian brain, microvessels may produce a special microenvironment and then influence the characteristics of neural stem cells (NSCs). This work aims to compare the effects of cerebral microvascular endothelial cells (CMECs) and vascular endothelial growth factor (VEGF) on the behaviors of NSCs. An in vitro model with co-cultured CMECs and NSCs was established by using transwell co-culture system. The expression of nestin and NF in the early stage of the co-culture, and NF in the later stage were explored by immunostaining. Results demonstrate that both CMECs and VEGF stimulate the proliferation of NSCs and inhibit the differentiation. After removing CMECs or VEGF from the co-culture system, a further culture in medium for the differentiation of NSCs was performed. The percentage of NF-positive cells in the CMECs co-culture group is the highest, suggesting the improved neuronal differentiation of NSCs by CMECs.

Polycrystalline diamond (PCD)-coated tungsten carbide (WC) is a high-performance cutting tool material widely used in industry for cutting or ultra-precision cutting of advanced engineering materials, but it is difficult to machine using conventional machining techniques owing to its high hardness. However, wire electrical discharge machining (wire EDM) offers advantages in machining PCD-coated WC in terms of process reliability and flexibility to machine different shapes. To relieve pollution issues associated with traditional wet wire EDM, the present study investigated the feasibility of near-dry wire EDM of PCD-coated WC cutting tool material using deionized water and air mist. A parametric study on spark cycle and spark duration was carried out, and the effect of these two important wire EDM process parameters on machining performance in terms of material removal rate (MRR) and surface roughness ( Ra) was analysed. Compared with conventional wet wire EDM, near-dry wire EDM consistently produces better Ra values on the PCD-coated WC workpiece, but near-dry wire EDM produces lower MRR than wet wire EDM under some conditions.

Alkyl glucosides were synthesized by the reaction of Leuconostoc mesenteroides dextransucrase with sucrose and various alcohols. Alkyl alpha-D-glucosides were obtained with a yield of 30% (mol/mol) with primary alcohols, but secondary alcohols or tertiary alcohols gave yields below 5%. The optimal yield was 50% using 1-butyl alpha-D-glucoside with 0.9 M 1-butanol. The acceptor products of methanol or ethanol were confirmed as methyl alpha-D-glucopyranoside and ethyl alpha-D: -glucopyranoside via MALDI-TOF MS and NMR analysis. Thus, methyl or ethyl alpha-D-glucoside constituted half the emulsification activities of Triton X-100 as commercially available surfactants.

A series of novel 4-(4-(5-methyl-3-arylisoxazol-4-yl)thiazol-2-yl)piperidyl carboxamides and thiocarboxamides were synthesized as potential lead compounds of inhibitors targeting D1 protease in plants. These compounds were designed on the basis of a D1 protease inhibitor hit structure identified by homology modeling and virtual screening. The syntheses of these compounds were accomplished via a four-step procedure including the isoxazole ring formation, alpha-bromination of acetyl group, thiazole ring formation, and carboxamide/thiocarboxamide attachment. The in vivo herbicidal activity tests show that most compounds possess moderate to good herbicidal activities. The enzyme activity of one compound against the native spinach D1 protease exhibits a competitive inhibition. The results suggest that these compounds are indeed potential inhibitors for targeting D1 protease in plants.

The experimental results for silicon carbide (SiC) wheel with fine grit size grinding of silicon carbide (Si3N4) revealed that the grinding parameters affect not only the ground silicon nitride surface roughness, but also the degree of surface damage. There exists complex non-linear relationship between the grinding parameters and surface quality. Better surface roughness doesn’t surely mean less surface damage. A method of prediction of grinding quality based on support vector regression is then presented according to the condition of small samples. The result shows the prediction accuracy based on this method is obviously higher than neural network, which provides an effective way for optimizing the grinding parameters to ensure the grinding quality as well as grinding efficiency while grinding of silicon carbide using conventional abrasive.

In order to produce N44SH and N40UH high-grade magnet, both strip casting and hydrogen decrepitation processes were adopted, and the subsequent milling, sintering and heat-treatment processes were improved. The density of N40UH magnet is risen up to 7.65 g/cm c . Weight loss rate after the Highly Accelerated Stress Test (HAST) for 48 hours is less than 0.2 mg/cm b . The microstructure of magnet shows that the grain size is between 5 to 8 μm. The rich-Nd phase is fine and uniform, the pinhole is very few, such magnet can be used for wind power generator after phosphoruster treatment directly, and it can also be used in hydrogen atmosphere of hybrid car after surface treatment.

Twenty-seven Porphyra lines, including lines widely used in China, wild lines and lines introduced to China from abroad in recent years, were screened by random amplified polymorphic DNA (RAPD) technique with 120 operon primers. From the generated RAPD products, 11 bands that showed stable and repeatable RAPD patterns amplified by OPC-04, OPJ-18 and OPX-06, respectively were scored and used to develop the DNA fingerprints of the 27 Porphyra lines. Moreover, the DNA fingerprinting patterns were converted into computer language expressed with two digitals, 1 and 0, which represented the presence (numbered as 1) or absence (numbered as 0) of the corresponding band, respectively. Based on the above results, computerized DNA fingerprints were constructed in which each of the 27 Porphyra lines has its unique fingerprinting pattern and can be easily distinguished from others. Software named PGI (Porphyra germplasm identification) was designed for identification of the 27 Porphyra lines. In addition, seven specific RAPD markers from seven Porphyra lines were identified and two of them were successfully converted into SCAR (sequence characterized amplification region) markers. The developed DNA fingerprinting and specific molecular markers provide useful ways for the identification, classification and resource protection of the Porphyra lines.

Abstract Plant-parasitic nematodes are distributed worldwide and affect a broad range of important agronomic plant species. Chitinolytic bacteria were evaluated as potential biological control agents of the root-knot nematode, Meloidogyne incognita, on tomato. After transplantation of seedlings into pots containing soil amended with chitin compost, chitin broth, or respective controls, soil was inoculated with nematode eggs and infective second-stage juveniles (J2). Samples taken at 4, 6 and 8 weeks after inoculation indicated that fresh weights of plants did not vary between treatments or between treatments and controls. The gall index was lower in the plants grown in the chitin-amended soil at each time point. Activities of soil chitinase and β-1,3-glucanase were greater in those soils amended with chitin compost and chitin broth. Gall index of tomato root was negatively correlated with soil chitinase activity. Activities of tomato root chitinase and β-1,3-glucanase were higher in plants growing in non-chitin-amended soil at 6 and 8 weeks after nematode infestation. Chitinase activity in tomato root was positively correlated with the gall index of tomato root. The results indicate the potential of chitinase producing bacteria to alleviate nematode parasitism in important vegetable crops.

We report a facile and efficient strategy for the preparation of graphene-Ag2Se nanocomposites. The reduction of graphene oxide (GO) to graphene was accompanied by the formation of Ag2Se particles on the surface of graphene nanosheets. The results of X-ray diffraction (XRD) and Fourier transform infrared (FTIR) confirmed the efficient reduction of GO. Transmission electron microscopy (TEM) images of the sample revealed the morphology of the architecture of graphene-Ag2Se hybrid. Photoluminescence (PL) measurement was further employed to study the optical properties of the nanocomposites. The results show that the graphene-Ag2Se nanocomposites can be prepared by a simple hydrothermal method, and Ag2Se particles are well distributed on the graphene nanosheets.

Optical second harmonic generation (SHG) has been used to analyze the ULSI process, including low energy ion-implantation and rapid thermal annealing (RTA). The projected range of the low energy ion-implantation is approximately 20 nm, which results in a region that is transformed into an amorphous state by high dose ion-implantation, and the surface layer region is recrystallized during RTA process. The symmetrical point group of the Si (1 1 1) surface layer is 3 mm, which differs from that of centrosymmetric bulk Si which has no dipole contribution. The reflected SHG (RSHG) of the Si (1 1 1) is contributed mainly from the surface dipole and surface quadrupole. A plot of RSHG versus azimuthal angle reveals information on the recrystallization of amorphous silicon during RTA. The optimization of RTA step is determined by RSHG, which is shown to be suitable for analyzing the ULSI process.

Dibenz[ a,h ]anthracene (DB[ a,h ]A) and 7-nitrodibenz[ a,h ]anthracene (7-NDB[ a,h ]A) induced liver tumors when administered to neonatal B6C3F 1 mice. For protooncogene analysis, RNA was isolated from each of the liver tumors from treated mice and reverse-transcribed into cDNA. Portions of the K- and H- ras protein coding sequences were then amplified and analyzed for DNA sequence alterations. DB[ a,h ]A-induced liver tumors had a 100% (23/23) frequency of ras -protooncogene mutation, with 83% (19/23) occurring at the first base of K- ras codon 13 and resulting in G GC M C GC transversion; the remaining 17% (4/23) of the mutations were located at the second base of H- ras codon 61. In contrast, only four of nine (44%) of 7-NDB[ a,h ]A-induced liver tumors had ras -protooncogene mutations, with two each at K- ras codon 13 and H- ras codon 61. Combined with previous observations, the results indicate that the nitro substituent perpendicular to the aromatic moiety alters the chemical-induced protooncogene ...

Background and objective Interleukin (IL)-25 has been shown to play an important role in the pathogenesis of chronic rhinosinusitis with nasal polyps. Nasal polyps are associated with chronic inflammation of the mucous membranes in the paranasal sinuses and are involved in extracellular matrix (ECM) accumulation. The aim of this study is to evaluate the effects of IL-25 on myofibroblast differentiation, ECM production and the expression of matrix metalloproteinases in nasal polyp derived fibroblasts (NPDFs) and to determine the molecular mechanism underlying these processes. Materials and methods A total of 40 patients were enrolled in this study for Immunofluorescence studies. Expression of IL17 receptor B was evaluated by real time reverse transcription polymerase chain reaction (PCR) in NPDFs. NPDFs were stimulated with IL-25 for 48 h in the presence or absence of mitogen-activated protein kinase (MAPK) and NF-κB inhibitors or small interfering RNAs (siRNA). The protein levels of fibrosis active mediators were examined using western blotting. Fibroblast migration was evaluated with a scratch assay. The total collagen amount was analyzed with the Sircol collagen assay. Results IL-25 induced α-SMA, fibronectin, and MMP-1 and -13, which were dependent on IL-17RB. IL-25 also induced activation of NF-κB and mitogen-activated protein kinase (MAPKs). By using the specific inhibitor of ERK, p38, JNK and NF-κB (U, SB, SP and Bay), we found that IL-25-induced expressions of α-SMA, fibronectin, and MMPs was regulated by the signaling pathways of MAPKs and NF-κB. IL-25 also induces α-SMA, fibronectin, and MMPs expression through IL-17RB-dependent pathways in NPDFs. The increased migration ability induced by IL-25 was suppressed by the specific inhibitors of MAPKs and NF-κB. Conclusion Our data indicate that IL-25 induced myofibroblast differentiation, fibronectin production, and MMP-1 and -13 expressions through the signaling pathways of MAPKs and NF-κB. in NPDFs and increased expression of IL-25 were also involved in the pathogenesis of nasal polyposis by affecting nasal fibroblasts in chronic rhinosinusitis with nasal polyps.

Human papillomavirus type 16 (HPV-16) infection is positively associated with cervical cancer, whereas adeno-associated virus (AAV) infection is negatively associated with this same cancer. In earlier studies these two virus types have been shown to directly interact, with AAV inhibiting or enhancing papillomavirus functions depending upon the specific circumstances. One defined interaction between these two viruses is the ability of the AAV Rep78 major regulatory protein to inhibit gene expression of the E6 promoter of BPV-1 (bovine papillomavirus type 1) and HPV types 16 and 18. As Rep78 is a DNA binding transcription factor, we considered whether Rep78 might bind HPV-16 DNA. Here, Rep78 is demonstrated to bind a 44-base pair region (nucleotides 14-56) within the HPV-16 p97 promoter using the electrophoretic mobility shift assay. This region is important for HPV-16 because it includes functional Sp1 and E2 protein binding motifs as well as part of the origin of replication. Furthermore, two Rep78 amino acid substitution mutants, at positions 77 or 64-65, were identified that did not recognize p97 DNA. Both of these Rep78 mutants were found to be defective for inhibition of p97 promoter activity in HeLa and T-47D nuclear extracts in vitro, in a transient chloramphenicol acetyltransferase assay, as well as defective for full inhibition of HPV-16-directed focus formation. These data, taken together, strongly suggest that the Rep78-p97 promoter interaction is at least partially responsible for Rep78-mediated inhibition of HPV-16. Finally, the finding that Rep78 specifically recognizes p97 DNA is surprising because the p97 promoter region contains no GAGC motifs, the core motif for Rep78 recognition. These data suggest that the p97 promoter may represent a new prototypical DNA target type for Rep78.

1-, 3-, and 6-nitrobenzo[a]pyrene (nitro-BaP) are environmental contaminants that can be metabolized to genotoxic derivatives by either nitroreduction or ring-oxidation. In this study, we examined the types of mutations produced by the primary nitroreduced metabolites, 1-, 3-, and 6-nitroso-BaP (NO-BaP) in the hprt gene of Chinese hamster ovary cells. RNA from 6-thioguanine-resistant mutants was reverse-transcribed to cDNA and the hprt coding sequence was amplified and sequenced. The mutational patterns produced by the three compounds exhibited extensive similarities: 1) base pair substitutions accounted for 67% (28/42) of 1-NO-BaP, 51% (26/51) of 3-NO-BaP, and 50% (11/22) of 6-NO-BaP mutations; 19-36% of the mutations were exon deletions and 14-18% were frameshifts; 2) most (64-84%) of the simple base pair substitutions occurred at G:C, mainly G:C-->T:A and G:C-->C:G transversions; 3) 98% (46/47) of the simple base pair substitutions at G:C had the mutated dG on the non-transcribed strand and 81% (38/47) were located with the mutated dG flanked 3' by at least one purine; and 4) most simple base pair substitutions (48/62, 77%) occurred in exons 2, 3, and 8 of the hprt gene. Although there were no significant differences among the mutation profiles of the NO-BaPs, a significant difference did exist between the mutation pattern produced by 3-NO-BaP and the mutation pattern previously determined for the ring-oxidized product of 3-nitro-BaP metabolism, trans-7,8-dihydroxy-anti-9,10-epoxy-7,8,9, 10-tetrahydro-3-nitrobenzo[a]pyrene. This observation indicates that differences in the structures of closely related adducts can be important enough to have an effect on mutation profiles.

The chemoenzymatic process involving biocatalytic resolution of rac-2-carboxyethyl-3-cyano-5-methylhexanoic acid ethyl ester (CNDE, 1) has been the most competitive and attractive route for pregabalin. A new esterase-producing strain ZJB-09203, which exhibited high hydrolytic activity, excellent enantioselectivity, and diastereoselectivity towards CNDE, has been successfully isolated from soil samples with a pH indicator agar plate method. The isolate was identified as Morgarella morganii by the ATB system (ID 32 GN) and the 16S rDNA sequence. In order to suppress product inhibition during enzymatic hydrolysis of CNDE, an adsorptive biocatalytic process was developed by utilizing anion-exchange resin D201 as adsorbent for selective removal of (3S)-2-carboxyethyl-3-cyano-5-methylhexanoic acid (2) from the reaction medium. This approach allowed the substrate loading to be increased up to 1.5 M and the chiral intermediate 2 was produced in 682 mM, 45.3 % conversion, and 95 % ee. These results imply that M. morganii ZJB-09203 esterase is a promising biocatalyst in the development of chemenzymatic manufacturing process for pregabalin.

We have characterized the DNA adducts of a series of nitro-polycyclic aromatic hydrocarbons (nitro-PAHs) formed under anaerobic conditions in vitro. Although the DNA adducts of nitro-PAHs formed through enzymatic nitroreduction are generally the C8-deoxyguanosyl adducts, nitroreduction of 1-nitrobenzo[a]pyrene (1-nitro-BaP), 3-nitro-BaP, and two derivatives in the presence of calf thymus DNA resulted in the N 2-deoxyguanosyl adducts with the deoxyguanosyl moiety remote from the reaction site. Two DNA adducts of this type were also formed from 1-nitropyrene as minor products. These DNA adducts were formed from anaerobic incubation with rat and mouse liver microsomes and cytosols in the presence of calf thymus DNA. Our results suggest that biological formation of this type of DNA adduct is independent of the enzymatic system, but dependent on the geometric structure and/or electronic features of the nitro-PAH molecules.

The tumorigenicity of 7-chlorobenz[α]anthracene (7-C1-BA), an environmental contaminant, and 7-bromobenz[α]anthracene (7-Br-BA) was determined in the male B6C3F 1 newborn mouse. Mice receiving 7-C1-BA and 7-Br-BA by i.p. injections at a dose of 1600 nmol per mouse on 1, 8, and 15 days after birth developed 92 and 96% hepatocellular adenomas, and 100 and 83% hepatocellular carcinoma, respectively. Metabolism by liver microsomes of 15-day-old mice each produced the corresponding trans -3,4-dihydrodiol. Analysis by 32 P-postlabeling/HPLC indicated the presence of DNA adducts derived from 7-C1-BA trans -3,4-dihydrodiol and 7-Br-BA trans -3,4-dihydrodiol. Our results indicate that both 7-C1-BA and 7-Br-BA are potent carcinogens and that bay-region diol epoxides are the ultimate metabolites that lead to DNA adduct formation and tumor initiation.

Both 1- and 3-nitrobenzo[a]pyrene (nitro-8aP) are environmental contaminants, potent mutagens in Salmonella, and moderate mutagens in Chinese hamster ovary (CHO) cells. The mutagenicity of their oxidized metabolites, trans-7,8-dihydroxy-anti-9,10-epoxy-7,8,9,10-tetrahydro-1-nitrobenzo[a]pyrene (1-nitro-Bap-DE) and trans-7,8-dihydroxy-anti-9,10-epoxy-7,8,9,10-tetrahydro-3-nitrobenzo[a]pyrene (3-nitro-Bap-DE), together with trans-7,8-dihydroxy-anti-9,10-epoxy-7,8,9,10-tetrahydrobenzo[a]pyrene (BaP-DE), was determined in CHO-K1 cells, and the resulting mutations at the hprt locus were characterized by polymerase chain reaction (PCR) amplification of reverse-transcribed hprt mRNA, followed by DNA sequence analysis. The mutant frequencies, in mutants/106 clonable cells, at 30 and 100 ng/ml, were BaP-DE, 248 and 456; 1-nitro-BaP-DE, 68 and 260; 3-nitro-BaP-DE, 81 and 232, respectively. In general, the three diolepoxides exhibited similar mutational spectra: 1) 64% (23/36 sequenced mutants) of BaP-DE, 53% (19/36) of 1-nitro-BaP-DE, and 64% (23/36) of 3-nitro-BaP-DE mutants resulted from simple base pair substitution, with the predominant mutation being GT transversion; 2) 90%, 100%, and 100% of mutations at G:C had the mutated dG on the nontranscribed DNA strand; and 3) about one quarter of the mutants produced by each mutagen had one or more PCR products with partial or complete exon deletions. The mutagens induced few frameshifts or complex mutations. Among the differences in mutational specificity for the three diolepoxides, the proportion of substituted dGs with 3′ purines was significant (P 0.05). Also, high proportions of BaP-DE and 3-nitro-BaP-DE base pair substitutions at G:C occurred in DNA sequence contexts of 5′-GG-3′, 5′-GGA-3′, and 5′-TGGA-3′, while the proportions of 1-nitro-BaP-DE mutants in these contexts were often lower. The results indicate that nitro substitution at C1 or C3 of BaP-DE reduces mutational potency in CHO cells and appears to have only subtle effects upon the mutational pattern in the hprt gene. © 1996 Wiley-Liss, Inc.

32 P-Postlabeling-HPLC is a highly sensitive analytical method for identification of chemical-modified DNA adducts isolated from samples obtained from experimental animals or humans exposed to carcinogenic chemicals. To determine optimal 32 P-postlabeling-HPLC conditions for efficient separation, we report here the use of ten diol-epoxide-modified 3′,5′-bisphosphate deoxynucleotides derived from benzo[ a ]pyrene (BaP), nitrated BaP, and related compounds. After testing ODS-modified, C 4 -modified, phenyl-modified, diphenyl-modified, and cyclodextrin-bonded reversed-phase HPLC columns, we found that the Vydac diphenyl-modified column can efficiently separate these 3′,5′-bisphosphate deoxynucleotides. The results suggest that 32 P-postlabeling-HPLC is a potentially useful methodology for detecting environmental carcinogens that can be metabolized to diol-epoxides. The relationships between the structures of anti -diol-epoxides and HPLC retention order are also discussed.

Effect of caloric restriction (CR) on metabolism of 7-fluorobenz-[a]anthracene (7-F-BA), 7-chlorobenz[a]anthracene (7-Cl-BA), 7-bromo-benz[a]anthracene (7-Br-BA), and 7-nitrobenz[a]anthracene (7-nitro-BA) by mouse liver microsomes was studied. The results indicate that CR can affect metabolism rate, modulate the regioselectivity and stereoselectivity of the cytochrome P-450 metabolizing enzymes, and lower the yield of the proximate mutagenic metabolites. These findings indicate that CR may in part modulate chemical carcinogenesis by altering the metabolic activation.

AIM: To investigate the effect of keratinocyte growth factor (KGF) gene therapy in acetic acid-induced ulcerative colitis in rat model. METHODS: The colitis of Sprague-Dawley rats was induced by intrarectal infusion of 1 mL 5% (v/v) acetic acid. Twenty-four hours after exposed to acetic acid, rats were divided into three experimental groups: control group, attenuated Salmonella typhimurium Ty21a strain (SP) group and SP strain carrying human KGF gene (SPK) group, and they were separately administered orally with 10% NaHCO3, SP or SPK. Animals were sacrificed and colonic tissues were harvested respectively on day 3, 5, 7 and 10 after administration. Weights of rats, colonic weight/length ratio and stool score were evaluated. Histological changes of colonic tissues were examined by hematoxylin and eosin (HE) staining method. The expression of KGF, KGF receptor (KGFR) and TNF-α were measured either by enzyme-linked immunosorbent assay or Western blotting. Immunohistochemistry was used to detect the cellular localization of KGFR and Ki67. In addition, superoxide dismutase (SOD) activity and malondialdehyde (MDA) contents in the homogenate were measured. RESULTS: Body weight and colonic weight/length ratio were declined in SPK group compared with SP and control groups (body weight: 272.78 ± 17.92 g vs 243.72 ± 14.02 g and 240.68 ± 12.63 g, P < 0.01; colonic weight/length ratio: 115.76 ± 7.47 vs 150.32 ± 5.99 and 153.67 ± 5.50 mg/cm, P < 0.01). Moreover, pathological changes of damaged colon were improved in SPK group as well. After administration of SPK strain, KGF expression increased markedly from the 3rd d, and remained at a high level till the 10th d. Furthermore, KGFR expression and Ki67 expression elevated, whereas TNF-α expression was inhibited in SPK group. In the group administered with SPK, SOD activity increased significantly (d 5: 26.18 ± 5.84 vs 18.12 ± 3.30 and 18.79 ± 4.74 U/mg, P < 0.01; d 7: 35.48 ± 3.35 vs 22.57 ± 3.44 and 21.69 ± 3.94 U/mg, P < 0.01; d 10: 46.10 ± 6.23 vs 25.35 ± 4.76 and 27.82 ± 6.42 U/mg, P < 0.01) and MDA contents decreased accordingly (d 7: 7.40 ± 0.88 vs 9.81 ± 1.21 and 10.45 ± 1.40 nmol/mg, P < 0.01; d 10: 4.36 ± 0.62 vs 8.41 ± 0.92 and 8.71 ± 1.27 nmol/mg, P < 0.01), compared with SP and control groups. CONCLUSION: KGF gene therapy mediated by attenuated Salmonella ameliorates ulcerative colitis induced by acetic acids, and it may be a safe and effective treatment for ulcerative colitis.