Novel strategy for depolymerization of avermectin fermentation residue to value-added amino acid product.

Avermectin fermentation residue (AFR) is rich in proteins, which can be depolymerized to value-added amino acids for in-plant reuse. The hydrochloric acid (HCl) hydrolysis is performed and investigated under different conditions, including HCl concentration, solid-liquid ratio, temperature, and time. The hydrolysis degree (HD) of 67.7% can be achieved. The empirical correlation of HD is established with a good practicability to control the HD and predict the experimental conditions. Solid-liquid reaction is confirmed to be dominant during the hydrolysis process. There are 17 kinds of amino acids in the hydrolysate, benefiting the reuse. Avermectin is not detected in the hydrolysate and AFR, and the mass of AFR is reduced by 53.8 wt%. This work provides a novel strategy for the environmentally friendly treatment and meanwhile the resource recovery of AFR.

Enhanced Dissolution of 7-ADCA in the Presence of PGME for Enzymatic Synthesis of Cephalexin.

Enzymatic catalysis has been recognized as a green alternative to classical chemical route for synthesis of cephalexin (CEX). However, its industrial practice has been severely limited by the low productivity due to the low solubility of 7-amino-3-deacetoxycephalosporanic acid (7-ADCA) and high hydrolysis of D-phenylglycine methyl ester (PGME). In this work, the enhanced dissolution of 7-ADCA in the presence of PGME for efficient enzymatic synthesis of CEX was investigated. Results showed that the solubility of 7-ADCA in water could be improved by PGME. Moreover, supersaturated solution of 7-ADCA could be created in the presence of PGME by a pH shift strategy. The supersaturated solution of 7-ADCA possess good stability, which could be explained in terms of the inhibition of 7-ADCA precipitation due to the presence of PGME. The interaction between 7-ADCA and PGME is explored by spectroscopic determination and DFT analysis and the mechanism of enhanced dissolution of 7-ADCA in the presence of PGME is discussed and proposed. The feasibility of supersaturated solution of 7-ADCA for the enzymatic synthesis of CEX is evaluated. It was demonstrated that high conversion ratio (> 95.0%) and productivity (> 240.0 mmol/L/h) was obtained under a wide range of reaction conditions, indicating that the supersaturated solution system was highly superior to conventional homogeneous solution system. The information obtained in this work will be helpful to industrial production of CEX via enzymatic route.

Efficient enzymatic synthesis of cephalexin in suspension aqueous solution system.

An efficient method for the enzymatic synthesis of cephalexin (CEX) from 7-amino-3-deacetoxycephalosporanic acid (7-ADCA) and d-phenylglycine methyl ester (PGME) using immobilized penicillin G acylase (IPGA) as catalyst in a suspension aqueous solution system was developed, where the reactant 7-ADCA and product CEX are mainly present as solid particles. The effects of key factors on the enzymatic synthesis were investigated. Results showed that continuous feeding of PGME was more efficient for the synthesis of CEX than the batch mode. Under the optimized conditions, the maximum 7-ADCA conversion ratio of 99.3% and productivity of 200 mmol/L/H were achieved, both of which are much superior to the homogeneous aqueous solution system. Besides, IPGA still retained 95.4% of its initial activity after 10 cycles of enzymatic synthesis, indicating the excellent stability of this approach. The developed approach shows great potential for the industrial production of CEX via an enzyme-based route.

Novel tumor-targeting, self-assembling peptide nanofiber as a carrier for effective curcumin delivery.

The poor aqueous solubility and low bioavailability of curcumin restrict its clinical application for cancer treatment. In this study, a novel tumor-targeting nanofiber carrier was developed to improve the solubility and tumor-targeting ability of curcumin using a self-assembled Nap-GFFYG-RGD peptide. The morphologies of the peptide nanofiber and the curcumin-encapsulated nanofiber were visualized by transmission electron microscopy. The tumor-targeting activity of the curcumin-encapsulated Nap-GFFYG-RGD peptide nanofiber (f-RGD-Cur) was studied in vitro and in vivo, using Nap-GFFYG-RGE peptide nanofiber (f-RGE-Cur) as the control. Curcumin was encapsulated into the peptide nanofiber, which had a diameter of approximately 10-20 nm. Curcumin showed sustained-release behavior from the nanofibers in vitro. f-RGD-Cur showed much higher cellular uptake in αvß3 integrin-positive HepG2 liver carcinoma cells than did non-targeted f-RGE-Cur, thereby leading to significantly higher cytotoxicity. Ex vivo studies further demonstrated that curcumin could accumulate markedly in mouse tumors after administration of f-RGD-Cur via the tail vein. These results indicate that Nap-GFFYG-RGD peptide self-assembled nanofibers are a promising hydrophobic drug delivery system for targeted treatment of cancer.

Analysis of Mogroside V in Siraitia grosvenorii with micelle-mediated cloud-point extraction.

INTRODUCTION: Mogroside V is the main effective ingredient of Siraitia grosvenorii used as a natural sweet food as well as a traditional Chinese medicine. The sample pre-treatment prior to chromatographic analysis requires large amounts of toxic organic solvents and is time consuming. OBJECTIVE: To develop an effective and simple method for extracting and determining mogroside V of Siraitia grosvenorii. METHODS: Mogroside V was extracted and preconcentrated by micelle-mediated cloud-point extraction with nonionic surfactant isotridecyl poly (ethylene glycol) ether (Genapol® X-080). The obtained solutions containing mogroside V were analysed by high-performance liquid chromatography (HPLC) with ultraviolet (UV) detection. The chromatographic separation was performed on a C18 -column using gradient elution with acetonitrile and water at 203 nm. RESULTS: The cloud-point extraction yield was 80.7% while the pre-concentration factor was about 10.8. The limit of detection was 0.75 µg/mL and the limit of quantification was 2 µg/mL. The relative standard deviations for intra- and interday precisions of mogroside V were less than 8.68% and 5.78%, respectively, and the recoveries were between 85.1% and 103.6%. CONCLUSION: The HPLC-UV method based on micelle-mediated cloud-point extraction for determination mogroside V in Siraitia grosvenorii was environmentally friendly, simple and sensitive.

Effect of bone marrow mesenchymal stem cell transplant on synovial proliferation in rats with type II collagen-induced arthritis.

OBJECTIVES: To investigate the influence and mechanism of bone marrow mesenchymal stem cell transplant in the synovial proliferation of type II collagen-induced arthritis. MATERIALS AND METHODS: From the bone marrow of Sprague-Dawley rats, mesenchymal stem cells were isolated and expanded. Forty rats were randomly divided into 5 groups: normal control, early mesenchymal stem cell treatment, late mesenchymal stem cell treatment, early collagen-induced arthritis control, and late collagen-induced arthritis control. The mesenchymal stem cells and normal saline were injected through the tail vein, and the following parameters were observed: arthritis index, articular pathology changes, serum vascular endothelial growth factor level, tumor necrosis factor-?, and interluekin-17 levels as detected through stable enzyme-linked immunosorbent assay. RESULTS: The arthritis index and articular pathologic scores of the early and late treatment groups were lower compared with those of the control groups (P < .05). The arthritis index and articular pathologic scores of the late treatment group were lower than those of the early treatment group (P < .05). The levels of vascular endothelial growth factor, tumor necrosis factor-α, and interluekin-17 of the early and late treatment groups were significantly decreased compared with the collagen-induced arthritis control groups (P < .05), and these levels were positively correlated with the arthritis index and articular pathologic scores (P < .05). CONCLUSIONS: The transplant of mesenchymal stem cells in rats with collagen-induced arthritis can inhibit the proliferation of synovium, which may be attributed to the reduced expression of vascular endothelial growth factor, tumor necrosis factor-α, and interluekin-17.

Correlations of trace element levels in the diet, blood, urine, and feces in the Chinese male.

In order to explore the associations between trace elements in dietary intake and the other three biological media (blood, urine, or feces) and inter-element interactions among the latter, we simultaneously collected 72-h diet duplicates, whole blood, and 72-h urine and feces from 120 free-living healthy males in China. Correlations among the toxic (cadmium [Cd], lead [Pb]), and nutritionally essential (zinc [Zn], copper [Cu], iron [Fe], manganese [Mn], selenium [Se], iodine [I]) elements were evaluated using Spearman rank correlation analysis based on analytical data determined by inductively coupled plasma-mass spectrometry. Dietary Cd intakes were highly correlated with the fecal Cd and blood Cd levels. Inverse correlations were found for Fe-Cd and Fe-Pb in both diet versus blood and diet versus feces. Cd-Zn and Cd-Se were significantly directly correlated in the urine and feces. Cd-Se and Pb-Se were negatively correlated in blood. In addition, there existed an extremely significant association between urinary Se and urinary I. Moreover, the other two highly direct correlations were found for Se-Fe and for I-Fe in urine. Improved knowledge regarding their mutual associations is considered to be of fundamental importance to understand more the complex interrelationships in trace element metabolism.

First-principles investigation of Ag-doped gold nanoclusters.

Gold nanoclusters have the tunable optical absorption property, and are promising for cancer cell imaging, photothermal therapy and radiotherapy. First-principle is a very powerful tool for design of novel materials. In the present work, structural properties, band gap engineering and tunable optical properties of Ag-doped gold clusters have been calculated using density functional theory. The electronic structure of a stable Au(20) cluster can be modulated by incorporating Ag, and the HOMO-LUMO gap of Au(20-) (n)Ag(n) clusters is modulated due to the incorporation of Ag electronic states in the HOMO and LUMO. Furthermore, the results of the imaginary part of the dielectric function indicate that the optical transition of gold clusters is concentration-dependent and the optical transition between HOMO and LUMO shifts to the low energy range as the Ag atom increases. These calculated results are helpful for the design of gold cluster-based biomaterials, and will be of interest in the fields of radiation medicine, biophysics and nanoscience.

Expression and function of miRNA in postoperative radiotherapy sensitive and resistant patients of non-small cell lung cancer.

PURPOSE: To investigate the different miRNA expression profiles of postoperative radiotherapy sensitive and resistant patients of non-small cell lung cancer, explore their potential role and find some radio-sensitivity markers. MATERIALS AND METHODS: Thirty non-small cell lung cancer patients who have been treated by postoperative radiotherapy were selected and were divided into radiotherapy sensitive group and resistant group according to overall survival and local or distant recurrence rate. Expression profile of miRNA in these two groups was detected by a microarray assay and the results were validated by quantitative RT-PCR and Northern blot. At the molecular level, the effect of one differently expressed miRNA (miR-126) on the growth and apoptosis of SK-MES-1 cells induced by irradiation was examined. RESULTS: Comparing with resistant patients, five miRNAs (miRNA-126, miRNA-let-7a, miRNA-495, miRNA-451 and miRNA-128b) were significantly upregulated and seven miRNAs (miRNA-130a, miRNA-106b, miRNA-19b, miRNA-22, miRNA-15b, miRNA-17-5p and miRNA-21) were greatly downregulated in radiotherapy sensitive group. Overexpression of miRNA-126 inhibited the growth of SK-MES-1 cells and promoted its apoptosis induced by irradiation. The expression level of p-Akt decreased in miRNA-126 overexpression group. After treating with phosphoinositidyl-3 kinase (PI3K) constitutively activator (IGF-1) and inhibitor (LY294002), miRNA-126 overexpression had no significant effects on the apoptosis of SK-MES-1 cells. CONCLUSION: We found 12 differently expressed miRNAs in the radiotherapy sensitive and resistant non-small cell lung cancer samples. Moreover, our results showed miRNA-126 promoted non-small cell lung cancer cells apoptosis induced by irradiation through the PI3K-Akt pathway.

Expression of miRNA-130a in nonsmall cell lung cancer.

MicroRNAs are short regulatory RNAs that negatively modulate gene expression at the posttranscriptional level and are deeply involved in the pathogenesis of several types of cancer. The miRNA-130a has been shown to play a role in antagonizing the inhibitory effects of GAX on endothelial cell proliferation, migration and tube formation, and antagonizing the inhibitory effects of HoxA5 on tube formation in vitro. Here the authors show, for the first time, that miRNA-130a expression is increased in nonsmall cell lung cancer (NSCLC) tissues. Statistical analysis showed that overexpression of miRNA-130a was strongly associated with lymph node metastasis, stage of tumor node metastasis classification and poor prognosis. Moreover, there was a significant difference in miRNA-130a expression levels between smoking and nonsmoking patients. Multivariate Cox regression analysis showed that miRNA-130a was an independent prognostic factor for patients with NSCLC. Together, these data suggest that miRNA-130a may comprise a potential novel prognostic marker for this disease.

Element contents in organs and tissues of Chinese adult men.

This study was undertaken to provide reference values for relevant parameters of Chinese Reference Man. Eighteen kinds of major organ or tissue samples, including muscle, rib, liver, and so on, were obtained from autopsies of 68 healthy adult men living in four areas of China with different dietary patterns (Hebei, Shanxi, Sichuan, Jiangxi or Jiangsu provinces, including Shanghai City) who had just encountered sudden deaths. At the same time, whole blood samples were collected from 10 volunteers living in each of these areas. The concentrations of 60 elements in these samples were detected by using inductively coupled plasma mass spectrometry (ICP-MS), inductively coupled plasma-atomic emission spectrometry (ICP-AES), neutron activation analysis (NAA), fluorometry (FL), graphite furnace atomic absorption spectrometry (GF-AAS) techniques and necessary quality control (QC) measures. Based on obtained concentrations and reference values of these organ or tissue weights for Chinese Reference Man, the elemental burdens in these organs or tissues were estimated. As a summary report of a series of research studies for Chinese Reference Man, which included three steps (from 1996 to 2006), the concentrations of 60 elements in 18 main organs or tissues were determined and their elemental burdens in the organs or tissues and whole body were estimated. Furthermore, the organ or tissue distributions of some important elements for radiation protection were discussed. These results may provide more reliable and better representative bases than before for establishing related reference values of Chinese Reference Man and revising current reference values of International Commission on Radiological Protection (ICRP) Reference Man. These basic data will also be very valuable for many other applications in radiation protection and other scientific fields.

pH neutralization while succinic acid adsorption onto anion-exchange resins.

Succinic acid is a useful chemical and its purification from fermentation broth by ion-exchange resins has widely drawn attention. In this study, pH neutralization in the process of adsorption of succinic acid from model solutions and fermentation broth by anion-exchange resin NERCB 04 has been tested. Adsorption capacity of NERCB 04 was about 0.41 g succinic acid/g resin at concentrations of succinic acid in the range of 10-50 g/L in packed column. In the process of succinic acid removal, pH of the system could also be neutralized. The neutralizing ability of the resin as a neutralizing agent has also been studied in the model cycle system and in the real fermentation cycle process. It was found that NERCB 04 showed stable adsorption capacity and pH neutralization ability after each regeneration. A certain amount of anion-exchange resin could neutralize the low pH values (pH 2-5) and maintain the system around pH 7.0. This means the anion-exchange resins have the function of neutralizing reagent in the process of adsorbing succinic acid.

Elemental transfer from Chinese soil via the diet to the whole human body.

Based on results from recent studies of elemental dietary intake and organ or tissue content for adult Chinese men, quoted nationwide elemental concentrations in Chinese soil and newly published national average consumption of dietary foods, values of both transfer coefficients and discrimination factor (DF) for transfer from soil via the diet to both critical organs and the whole body have been calculated for important elements in radiation protection, including alkaline earths, alkali metals, rare earths and other related elements. These calculations have used both the United Nations Scientific Committee on the Effects of Atomic Radiation (UNSCEAR) model and the DF method. In the UNSCEAR model, the basic parameters used to describe the transport of radionuclides are the transfer coefficients P(ij), which describe the relationship of concentrations or other amounts between compartment i and the following compartment j, whereas the DF is the ratio between the transfer coefficients for one element and a chemically similar element. From a comparison of the transfer coefficients of different elements for a particular transport pathway, those for alkaline earths are generally speaking higher than those for halogen elements and alkali metals, whereas those for rare earth elements, U and Th are lower. Relative to Ca, the DFs of transfer from soil to diet and from diet to critical organs or the whole body for the other alkaline earth elements and Pb are all less than 1, the DFs for the other elements decrease with increasing or decreasing atomic number. For alkali metals, the DFs of transfer from diet to critical organs and the whole body seem to increase with increasing atomic number, but those from soil to diet decrease with increasing atomic number.

Elemental contents in organs and tissues of Chinese adult men.

OBJECTIVE: To provide basis of reference values for relevant parameters of Chinese Reference Man. METHODS: Eighteen kinds of major organ or tissue samples, including muscle, rib, liver, and so on, were obtained from 4 areas (Hebei, Shanxi, Jiangsu, and Sichuan provinces) with different dietary patterns in China in autopsy of 16 healthy adult men, who had just encountered sudden deaths. At the same time, whole blood samples were collected from 10 volunteers living in each of these areas. The concentrations of 56 elements in these samples were detected by using Inductively Coupled Plasma Mass Spectrometry (ICP-MS), Inductively Coupled Plasma Atomic Emission Spectrometry (ICP-AES), and Graphite Furnace Atomic Absorption Spectrometry (GF-AAS) techniques. Based on obtained concentrations and reference values of these organ or tissue weights for Chinese Reference Man, the relative elemental burdens in these organs or tissues as well whole body were also estimated. RESULTS: The concentrations of 56 elements in 18 main organs or tissues were determined all together and their elemental organ or tissue and whole body burdens were estimated. Furthermore, the distributions of important elements for radiation protection in these organs or tissues were emphatically discussed. CONCLUSION: By summing with past related results, the total results obtained from the series of research may provide more reliable and better representative basis of these reference values for Chinese Reference Man than before.

Effects of zinc, copper, and selenium on placental cadmium transport.

The objective of the present study was to evaluate the potential effects of zinc, copper, and selenium on placental cadmium transport. From November 2002 through January 2003, a total of 47 healthy pregnant women from Da-Ye City, Hubei Province in Central China participated in the study. Their age, parity, gestational age, pregnancy history, and lifestyle data were obtained by questionnaire interview. The placental, whole-blood, and cord blood levels of cadmium were determined by inductively coupled plasma mass spectrometer (ICP-MS), whole-blood zinc was measured by flame atomic absorption spectrometry (F-AAS), whole-blood copper by ICP-MS, and selenium was by atomic fluorescence spectrophotometry (AFS). The cord blood cadmium concentration (0.020-1.48 microg/L) was significantly lower than in maternal blood (0.80-25.20 microg/L, p<0.01). The placental cadmium concentration was from 0.082 to 3.97 microg/g dry weight. Multiple linear regression analysis indicated that lower levels of maternal blood copper were significantly associated with higher cadmium concentrations in cord blood. Placental cadmium in women with lower levels of maternal blood zinc was significantly higher than in those with normal zinc levels. The placental cadmium level in women with lower whole-blood selenium was significantly lower than in subjects with normal selenium levels. It was concluded that the essential elements copper, selenium, and zinc might significantly affect placental cadmium transport.

Effect of environmental exposure to cadmium on pregnancy outcome and fetal growth: a study on healthy pregnant women in China.

The objective of the present study was to evaluate the potential effect of environmental exposure to cadmium on pregnancy outcome and fetal growth. Normal pregnant women were selected from Da-ye city of Hubei province, a cadmium-polluted area, from November 2002 through January 2003. Whole blood of pregnant women, cord blood, and placenta were collected and cadmium levels were determined by inductively coupled plasma emission mass spectroscopy. Incidence rate of preterm labor (gestational age < or = 37 weeks) and neonatal asphyxia, neonatal birth height, and birth weight were compared between lower and higher cadmium exposure level groups. Whole blood cadmium of 44 mothers ranged from 0.80 to 25.20 microg/L. Cadmium concentration in maternal blood was significantly higher than that in cord blood (t = 11.44, P < 0.01). Placenta cadmium ranged from 0.084 to 3.97 microg/g dry weight. After adjustment for maternal age, history of gestation, abortion and lactation, Logistic regression analysis showed that there was no significant association between cadmium exposure levels and pregnancy outcome (premature labor or neonatal asphyxia). Multiple linear regression analysis showed that, cord blood cadmium level, but not maternal blood cadmium and placenta cadmium, was significantly negatively associated with neonatal birth height (t= -2.33, P < 0.05). Compared with lower cord blood cadmium level (< or = 0.40 microg/L), higher level of cord blood cadmium (>0.40 microg/L) was associated with 2.24cm decrease in neonatal birth height. There was no significant association between cadmium exposure and birth weight. It was concluded that environmental exposure to cadmium significantly lower neonatal birth height.