Construction of an alternative glycerol-utilization pathway for improved ß-carotene production in Escherichia coli.

Glycerol, which is an inevitable by-product of biodiesel production, is an ideal carbon source for the production of carotenoids due to its low price, good availability and chemically reduced status, which results in a low requirement for additional reducing equivalents. In this study, an alternative carbon-utilization pathway was constructed in Escherichia coli to enable more efficient ß-carotene production from glycerol. An aldehyde reductase gene (alrd) and an aldehyde dehydrogenase gene (aldH) from Ralstonia eutropha H16 were integrated into the E. coli chromosome to form a novel glycerol-utilization pathway. The ß-carotene specific production value was increased by 50% after the introduction of alrd and aldH. It was found that the glycerol kinase gene (garK), alrd and aldH were the bottleneck of the alternative glycerol metabolic pathway, and modulation of garK gene with an mRS library further increased the ß-carotene specific production value by 13%. Finally, co-modulation of genes in the introduced aldH-alrd operon led to 86% more of ß-carotene specific production value than that of the strain without the alternative glycerol-utilization pathway and the glycerol-utilization rate was also increased. In this work, ß-carotene production of E. coli was significantly improved by constructing and optimizing an alternative glycerol-utilization pathway. This strategy can potentially be used to improve the production of other isoprenoids using glycerol as a cheap and abundant substrate, and therefore has industrial relevance.

[Simultaneous determination of 7 benzodiazepines in human plasma by UHPLC-MS/MS].

The study was designed to develop the method for determination of 7 benzodiazepines concentration in human plasma. UHPLC-MS/MS was adopted to analyze plasma with protein precipitated by acetonitrile. Citalopram was used as an internal standard. Plasma samples were separated on CORTECS UHPLC C18 column with the mobile phase of aqueous solution (0.01% formic acid) - methanol (0.01% formic acid) at a flow rate of 0.3 m L·min(-1). Multiple reaction monitoring (MRM) mode was performed in combiation with electrospray ionization source operating in the positive ionization mode. The liner calibration curve of midazolam, nitrazepam, estazolam, clonazepam, lorazepam, triazolam and diazepam were obtained in the concentration range of 1.05-840 (r = 0.999 4), 2.06-824 (r = 0.998 1), 2.02-1 616 (r = 0.994 7), 6.18-2 472 (r = 0.997 9), 6.12-2 448 (r = 0.997 4), 3.02-2 416 (r = 0.990 2), 1.02-816 (r = 0.998 8) ng·m L(-1), respectively. The lowest detection limit were 0.02, 0.52, 0.51, 1.55, 0.77, 0.76, 0.02 ng·m L(-1), respectively. The RSD of inter-day and intra-day were less than 10.81%. The relative recovery was 81.46%-106.53%. The method was successfully applied to clinical analysis of blood samples from patients.

Safety and efficacy of postoperative adjuvant therapy with atezolizumab and bevacizumab after radical resection of hepatocellular carcinoma.

BACKGROUND: The effects of postoperative adjuvant therapy for high-risk recurrent hepatocellular carcinoma (HCC) in immunotherapy are still under investigation. This study evaluated the preventive effects and safety of postoperative adjuvant therapy, including atezolizumab, and bevacizumab, against the early recurrence of HCC with high-risk factors. METHODS: The complete data of HCC patients who underwent radical hepatectomy with or without postoperative adjuvant therapy after two-year follow-up were analyzed retrospectively. The patients were divided into high-risk or low-risk groups based on HCC pathological characteristics. High-risk recurrence patients were divided into postoperative adjuvant treatment and control groups. Due to the difference in approaches in postoperative adjuvant therapies, they were divided into transarterial chemoembolization (TACE), atezolizumab, and bevacizumab (T + A), and combination (TACE+T + A) groups. The two-year recurrence-free survival rate (RFS), overall survival rate (OS), and associated factors were analyzed. RESULTS: The RFS in the high-risk group was significantly lower than that in the low-risk group (P = 0.0029), and the two-year RFS in the postoperative adjuvant treatment group was significantly higher than that in the control group (P = 0.040). No severe complications were observed in those who received atezolizumab and bevacizumab or other therapy. CONCLUSION: Postoperative adjuvant therapy was related to two-year RFS. TACE, T + A, and the combination of these two approaches were comparable in reducing the early recurrence of HCC without severe complications.

Association of tumor necrosis factor-α gene promoter polymorphism at sites -308 and -238 with non-alcoholic fatty liver disease: a meta-analysis.

BACKGROUND AND AIM: Environmental and genetic factors play a role in the pathogenesis and natural history of non-alcoholic fatty liver disease (NAFLD). The objective of this study was to quantitatively evaluate the association between tumor necrosis factor (TNF)-α gene promoter polymorphism at sites -308 and -238 and NAFLD susceptibility. METHODS: We performed an extensive search of relevant studies and made a meta-analysis, including eight studies with 837 NAFLD cases and 990 controls in the association between TNF-α -308 polymorphism and NAFLD; and seven studies with 771 cases and 787 controls in TNF-α -238 polymorphism. RESULTS: The combined results showed that there was a significant difference in TNF-α-238 genotype distribution between NAFLD and control based on all studies (GA/AA vs GG [odds ratio = 2.06, 95% confidence interval = 1.58-2.69, P < 0.000,01]). However, the combined results based on all studies showed there was no evidence of association of TNF-α-308 genotype distribution between NAFLD cases and controls (GA/AA vs GG [odds ratio = 1.08, 95% confidence interval = 0.82-1.42, P = 0.60]). When stratifying for race, the significant results did not change materially compared with whole populations. CONCLUSION: This meta-analysis suggested that TNF-α gene promoter polymorphism at position -238 but not -308 might be a risk factor for NAFLD.

Reforming Magnet Waste to Prussian Blue for Sustainable Sodium-Ion Batteries.

Increasing generation of permanent magnet waste has resulted in an urgent need to preserve finite resources. Reforming these wastes as feedstock to produce renewables is an ideal strategy for addressing waste and energy challenges. Herein, our work reports a smart and sustainable strategy to convert iron in magnet wastes into Prussian blue analogues that can serve as cathode materials for sodium-ion batteries. Moreover, a method to control feed rates is proposed to generate high-quality materials with fewer [Fe(CN)6] vacancies at a feed rate of 3 mL min-1. The recycled Na1.46Fe[Fe(CN)6]0.85·â–¡0.15 shows low vacancies and excellent cycling stability over 300 cycles (89% capacity retention at 50 mA g-1). In operando, evidence indicates that high-quality Prussian blue allows fast sodium-ion mobility and a high degree of reversibility over the course of cycling, although with a three-phase-transition mechanism. This study opens up a future direction for magnet waste created with the expectation of being environmentally reused.

Engineering CrtW and CrtZ for improving biosynthesis of astaxanthin in Escherichia coli.

This study engineered ß-carotene ketolase CrtW and ß-carotene hydroxylase CrtZ to improve biosynthesis of astaxanthin in Escherichia coli. Firstly, crtW was randomly mutated to increase CrtW activities on conversion from ß-carotene to astaxanthin. A crtW* mutant with A6T, T105A and L239M mutations has improved 5.35-fold astaxanthin production compared with the wild-type control. Secondly, the expression levels of crtW* and crtZ on chromosomal were balanced by simultaneous modulation RBS regions of their genes using RBS library. The strain RBS54 selected from RBS library, directed the pathway exclusively towards the desired product astaxanthin as predominant carotenoid (99%). Lastly, the number of chromosomal copies of the balanced crtW-crtZ cassette from RBS54 was increased using a Cre-loxP based technique, and a strain with 30 copies of the crtW*-crtZ cassette was selected. This final strain DL-A008 had a 9.8-fold increase of astaxanthin production compared with the wild-type control. Fed-batch fermentation showed that DL-A008 produced astaxanthin as predominant carotenoid (99%) with a specific titer of 0.88 g·L-1 without addition of inducer. In conclusion, through constructing crtW mutation, balancing the expression levels between crtW* and crtZ, and increasing the copy number of the balanced crtW*-crtZ cassette, the activities of ß-carotene ketolase and ß-carotene hydroxylase were improved for conversion of ß-carotene to astaxanthin with higher efficiency. The series of conventional and novel metabolic engineering strategies were designed and applied to construct the astaxanthin hetero-producer strain of E. coli, possibly offering a general approach for the construction of stable hetero-producer strains for other natural products.

The Efficacy and Safety of High-dose Daptomycin in the Treatment of Complicated Skin and Soft Tissue Infections in Asians.

OBJECTIVE: To compare the efficacy and safety of standard-dose (SD) daptomycin with those of high-dose (HD) daptomycin in complicated skin and soft tissue infections (cSSTIs) in an Asian population. MATERIALS AND METHODS: Patients from three medical centers diagnosed with cSSTIs were screened in the clinical information system. Patients included in the analysis were divided into two groups: those who received daptomycin at doses ≥ 6 mg/kg (HD group) and those receiving 4 mg/kg (SD group). The demographics and clinical treatment information were analyzed. RESULTS: Overall, 155 patients were recruited, including 108 patients in the SD group and 47 patients in the HD group. The rate of healthcare-associated infections was higher in the HD group (61.70% vs. 37.04%), demonstrating a statistically significant difference (P = 0.005). Compared with the SD group, the HD group had statistically significant early clinical stabilization (72.34% vs 52.78%, P = 0.023). The results of the multivariate analysis indicated that HD daptomycin was an independent effector for early clinical stabilization (HR=0.394, P < 0.001). The rate of drug-related adverse events was equally distributed in the HD and SD groups (36.17% vs. 26.85%, P = 0.243). CONCLUSION: Compared with SD daptomycin, HD daptomycin increased the rate of early clinical stabilization in Asian patients with cSSTIs, whereas the incidence of adverse events did not increase.

[Chemical constituents of Momordica charantia L].

Momordica charantia L. is a vegetable widely cultivated around the world. Its fruits have been used in Asian countries as a folk medicine for the treatment of non-insulin-dependent diabetes mellitus. Here we report eight compounds isolated from the fruits of M. charantia. On the basis of NMR and MS spectroscopic analyses, these compounds were identified as momordicolide ((10E)-3-hydroxyl-dodeca-10-en-9-olide, 1), monordicophenoide A (4-hydroxyl-benzoic acid 4-O-beta-D-apiofuranosyl (1 --> 2)-O-beta-D-glucopyranoside, 2), dihydrophaseic acid 3-O-beta-D-glucopyranoside (3), 6,9-dihydroxy-megastigman-4,7-dien-3-one (blumenol, 4), guanosine (5), adenosine (6), uracil (7) and cytosine (8). Among them, 1 and 2 are new compounds. Compounds 3-5 were isolated from this plant for the first time.

[Effect of supplemented Taoren Chengqi decoction on NIT-1, a pancreatic beta-cell from a transgenic NOD/Lt mouse].

OBJECTIVE: To study the effect of Supplemented Taoren Chengqi decoction (STCD) on the secretion of insulin and proliferation of NIT-1. METHOD: The effect of STCD and the serum of rat after orally administrating of STCD on the secretion of insulin and proliferation of NIT-1 were studied. The proliferation of NIT-1 was measured by 3H-TdR incorporation and cell counting methods, while the secretion of insulin was measured from the cultured medium by the ultra sensitive rat insulin ELISIA kit. RESULT: Both the STCD and the serum of rat after orally administrating of STCD significantly could increased the secretion of insulin and proliferation of NIT-1. CONCLUSION: The treatment of the diabetic patients by STCD might be through with its improvement of secretion of insulin and proliferation on pancreatic beta-cell.

[Effect of beryllium on the morphology and chemical elements of cell membrane of Porphyromonas gingivalis].

PURPOSE: To evaluate the effect of beryllium (Be²âº) on the morphology and chemical elements on cell membrane of Porphyromonas gingivalis (P. gingivalis), thus to explore the microbiologic mechanisms of periodontal diseases. METHODS: P. gingivalis was put into the culture with different Be²âº concentrations and anaerobically cultured for 24 hours. The morphologic change of P. gingivalis was observed under microscope and scanning electronic microscope (SEM), and chemical elements of cell membrane were observed by X-ray energy dispersion spectrum (EDS). The data was statistically analyzed with SPSS13.0 software package. RESULTS: The morphology of P.gingivalis altered obviously at the concentration greater than 2.5 mg/L, which was manifested by the sharpness of border and depression on the surface. With the increased concentration of beryllium, the Na and Ca peak descended on the surface of P. gingivalis. CONCLUSIONS: Beryllium can interfere with the morphology of P. gingivalis, and lead to the changes of chemical elements on cell membrane of P. gingivalis, which may result in a disturbance in the microecologic balance of subgingival microbes and eventually contribute to periodontal diseases.

Meat intake and risk of inflammatory bowel disease: A meta-analysis.

BACKGROUND/AIMS: This meta-analysis is designed to determine the association between meat consumption and the risk of inflammatory bowel disease. MATERIALS AND METHODS: Search relevant literature published in PubMed, Cochrane before July 2015 without restrictions. Studies were included if relative ratios and the corresponding 95% confidence intervals of the risk of inflammatory bowel disease were reported with respect to meat consumption. RESULTS: Nine studies were included in this meta-analysis. Relative to those who did not or seldom eat meat, meat consumers had a significantly greater risk of inflammatory bowel disease (pooled relative ratio: 1.50, 95% confidence interval: 1.15-1.95). The funnel plot revealed no evidence for publication bias. CONCLUSION: Meat consumption may increase the risk of inflammatory bowel disease. Additional large prospective studies are warranted to verify this association.

Effect of siRNA on Wisp-1 gene expression, proliferation, migration and adhesion of mouse hepatocellular carcinoma cells.

OBJECTIVE: To study the inhibition effect of siRNA on the expression of Wisp-1 gene in Hca-F of mouse hepatocellular carcinoma cells strain and also its effect on the proliferation, migration and adhesion of hepatocellular carcinoma cells. METHODS: Three expression vectors of siRNA were constructed. Lipo2000 was employed to transfect Hca-F cells and Western blot was used to detect the inhibition effect of siRNA on the expression of Wisp-1 gene. Afterward, CCK8 was adopted to detect the effect of Wisp-1 siRNA on the proliferation of Hca-F cells; Annexin V-FITC/PI double staining flow cytometry was used to detect the effect of Wisp-1 siRNA on the apoptosis of Hca-F cells; Transwell was used to detect the effect of Wisp-1 siRNA on the migration of Hca-F cells. The in vitro cell adhesion kit was used to detect of Wisp-1 siRNA on the change in the components of extracellular matrix to which Hca-F cells adhered. Western blot was used to detect the activation of protein kinase B (AKT)/glycogen synthase kinase-3ß pathway and the expression of downstream target protein p53 and matrix metalloproteinases-2. RESULTS: The siRNA showed interference effect on the expression of Wisp-1 gene. Compared with the control group, after being transfected to cells, Wisp-1 siRNA could significantly inhibit the proliferation, migration and adhesion of Hca-F cells and also promote the cell apoptosis, which was related to the down-regulated phosphorylation of AKT and glycogen synthase kinase-3ß and the expression of p53 and matrix metalloproteinases-2 (P < 0.05). CONCLUSIONS: The inhibition of Wisp-1 expression can reduce the proliferation, migration and adhesion of mouse hepatocellular carcinoma cells, which is related to the AKT/glycogen synthase kinase-3ß pathway. Wisp-1 gene may be the potential target to cure the hepatocellular carcinoma.

[Factors related to HIV-associated lipodystrophy among patients on antiretroviral therapy].

UNLABELLED: To describe the distribution and related risk factors of lipodystrophy (LD)among AIDS patients treated with antiretroviral drugs. METHODS: A cross-sectional study was performed on 261 AIDS patients treated with antiretroviral drugs. All the subjects were followed in the Center for Disease Control and Prevention of two counties in northern Anhui province from May 25 to 30, 2012. Data related to demography, physical examination, history of antiretroviral treatment, HIV plasma viral load, and CD4 + T cell count were collected. Clinical examination was based on an assessment of changes in face, legs, arms, buttocks(peripheral sites), back, chest, neck or abdomen or change in waist size (central sites)as quoted by the clinicians. RESULTS: LD was observed in 147 (56.3%) patients. The differences of age , gender, quality of sleep, weight and time of treatment between LD and non-lipodystrophy (NLD)groups were statistically significant (P < 0.05). Results from the Multivariate logistic regression analysis showed that the risk of women suffering from LD was 1.894 times of thd males (95%CI:1.075-3.338). The risk of those with LD showed an 1.448-fold increase regarding the time of treatment for each additional year (95%CI:1.267-1.654). Patients with poor quality of sleep were prone to LD with 11.901 times more than those with good quality of sleep (95%CI:2.701-52.441). CONCLUSION: LD was commonly observed in AIDS patients who were under antiretroviral therapy. Gender, tine of treatment and the quality of sleep appeared the main factors related to the results of observation.

Cucurbitane triterpenoids from Momordica charantia.

Three new cucurbitane-type triterpenoid saponins, 23-O-beta-D-allopyranosyl-5beta,19-epoxycucurbita-6,24-diene-3beta,22(S),23(S)-triol-3-O-beta-D-glucopyranoside (1), 23-O-beta-D-allopyranosyl-5beta,19-epoxycucurbita-6,24-diene-3beta,22(S),23(S)-triol-3-O-beta-D-allopyranoside (2), and 23-O-beta-D-allopyranosyl-5beta,19-epoxycucurbita-6,24-diene-3beta,19(R), 22(S),23(S)-tetraol-3-O-beta-D-allopyranoside (3), named momordicoside M, N, and O, respectively, along with one known saponin momordicoside L (4), were isolated from the fresh fruits of Momordica charantia. The structures of these saponins were elucidated on the basis of chemical properties and spectral data.