Adult ; Aged ; Female ; Humans ; Male ; Middle Aged ; Nursing Staff, Hospital ; psychology ; Surveys and Questionnaires

Lysostaphin is highly effective on eliminating methicillin resistant Staphylococcus aureus (MRSA). In order to achieve controlled release of lysostaphin, a biocompatible drug carrier is needed. Hydroxyapatite/chitosan (HA/CS) composites were chosen to carry lysostaphin and sample composites with different weight ratios of HA to CS, including 80/20, 70/30, 60/40, and 40/60, were prepared. Multiple analyses were performed to determine the structural and physicochemical properties of the composites, including scanning electron microscopy, X-ray diffraction and Fourier transform infrared spectroscopy. We immersed HA/CS composites loaded with 1 wt% lysostaphin to test in vitro release activity and cultured MC3T3-E1 cells to carry out biocompatibility test. The result of the release behavior of the composites revealed that the controlled release of lysostaphin from 60/40 HA/CS composites was the highest release rate of (87.4 ± 2.8)%, which lasted for 120 hours. In biocompatibility testing, MC3T3-E1 cells were able to proliferate on the surface of these composites, and the extract liquid from the composites could increase the growth of the cells. These results demonstrate the controlled release of lysostaphin from HA/CS composites and their biocompatibility, suggesting the potential application of these composites to bone injury and infection applications.

Lysostaphin is highly effective on eliminating methicillin resistant Staphylococcus aureus (MRSA). In order to achieve controlled release of lysostaphin, a biocompatible drug carrier is needed. Hydroxyapatite/chitosan (HA/CS) composites were chosen to carry lysostaphin and sample composites with different weight ratios of HA to CS, including 80/20, 70/30, 60/40, and 40/60, were prepared. Multiple analyses were performed to determine the structural and physicochemical properties of the composites, including scanning electron microscopy, X-ray diffraction and Fourier transform infrared spectroscopy. We immersed HA/CS composites loaded with 1 wt% lysostaphin to test in vitro release activity and cultured MC3T3-E1 cells to carry out biocompatibility test. The result of the release behavior of the composites revealed that the controlled release of lysostaphin from 60/40 HA/CS composites was the highest release rate of (87.4 ± 2.8)%, which lasted for 120 hours. In biocompatibility testing, MC3T3-E1 cells were able to proliferate on the surface of these composites, and the extract liquid from the composites could increase the growth of the cells. These results demonstrate the controlled release of lysostaphin from HA/CS composites and their biocompatibility, suggesting the potential application of these composites to bone injury and infection applications.
3T3 Cells ; Animals ; Biocompatible Materials ; Chitosan ; chemistry ; Delayed-Action Preparations ; Drug Carriers ; chemistry ; Durapatite ; chemistry ; Lysostaphin ; pharmacology ; Materials Testing ; Methicillin-Resistant Staphylococcus aureus ; Mice ; Microscopy, Electron, Scanning ; X-Ray Diffraction

Yeast strains with improved ethanol yield are important for efficient bioconversion of lignocellulosic biomass for fuel ethanol.Candida shehatae CICC1766 was adapted to 4%(v/v)ethanol,and then subjected to UV mutagenesis.One respiration deficient mutant Rd-5 with improved xylose fermentation capability was selected.Protoplasts of Rd-5 were inactivated by UV treatment,followed by the PEG-mediated protoplast fusion with a Saccharomyces cerevisiae strain with good ethanol-fermenting capability.The xylose fermenting capability of the fusants was investigated,and the fusant F6 demonstrated good ethanol fermentation performance,producing 18.75g/L ethanol from 50g/L xylose with an ethanol yield of 0.375 or 73.4% of its theoretical value of 0.511.Comparing with its parent Candida shehatae strain,the ethanol yield of F6 was increased by 28%.

Ethanol tolerance of self-flocculating yeast SPSC01 was studied in a 3-L bioreactor under fed-batch culture. Yeast floc populations with the average sizes around 100, 200, 300, and 400 microm were obtained by adjusting the mechanical stirring rates of the fermentation system. When subjected to 20% (V/V) ethanol shock for 6 h at 30 degrees C, the remained cell viability was 3.5%, 26.7%, 48.8% and 37.6% for the aforementioned four floc populations, respectively. The highest ethanol yield 85.5% was achieved for the 300 microm flocs, 7.2% higher than that of the 100 microm flocs. The amounts of trehalose and ergosterol (including free ergosterol and total ergosterol) were positively correlated with the average size distributions from 100 to 300 microm. However, in the 400 microm flocs, the content of trehalose and ergosterol decreased, which coincided with its reduced ethanol tolerance compared to that of the 300 microm flocs. Furthermore, when subjected to 15% (V/V) ethanol shock at 30 degrees C, the equilibrium nucleotide concentration and plasma membrane permeability coefficient(P') of the 300 microm flocs accounted for only 43% and 52% respectively of those of the 100 microm and 200 microm populations. The effect of floc size distribution on the ethanol tolerance of the self-flocculating yeast strain SPSC01 was closely related to plasma membrane permeability. An optimal floc size distribution with the highest ethanol tolerance and ethanol production level could be obtained by controlling mechanical stirring speed of the bioreactor, which provides basis for the process optimization of fuel ethanol production using this self-flocculating strain.
Bioreactors ; microbiology ; Drug Tolerance ; Ergosterol ; biosynthesis ; Ethanol ; metabolism ; pharmacology ; Fermentation ; Flocculation ; Industrial Microbiology ; methods ; Particle Size ; Trehalose ; biosynthesis ; Yeasts ; drug effects ; growth & development ; metabolism

AIM:To evaluate specific metabolomics profiles in the serum of patients with chronic mountain sickness (CMS) and to explore the potential metabolic biomarkers in the native Tibetans living on the Qinghai-Tibet Plateau.METHODS:A gas chromatography time-of-flight mass spectrometry (GC-TOF-MS) approach as a metabolomics technique was used to evaluate metabolic differences.The native Tibetan CMS patients (n =10) and healthy Tibetan controls (n =10) were enrolled from YuShu in Qinghai province in this study.The serum samples were collected and analyzed by GC-TOF-MS coupled with a series of multivariate statistical analyses such as principal component analysis (PCA),partial least squares discriminant analysis (PLS-DA) and orthogonal partial least squares discriminant analysis (OPLS-DA).RESULTS:The intergroup differences between CMS patients and control subjects have been observed.A list of differential metabolites and several top altered rnetabolic pathways have been identified.The levels of fumaric acid,an intermediate in the tricarboxylic acid (TCA) cycle,and inosine were highly upregulated in the CMS patients,suggesting a greater effort to hypoxic adaptation in high elevation area.Other differential metabolites,such as methyl phosphate,2-ketoadipate,lyxose and phytanic acid were also identified.Importantly,the differential metabolites possessed higher area under the ROC curve (AUC) values,indicating an excellent clinical ability for the prediction of CMS.Increased levels of amino acids (isoleucine,glycine,serine,L-cysteine,citrulline and trimethyllysine) were detected in CMS group,yet significantly decreased levels of sulfuric acid,oxamic acid,lyxose and glutamine were also detected in CMS group than those in control group.At the same time,the levels of ribose and glucose-1-phosphate were markedly elevated in CMS group (P ＜ 0.05).CONCLUSION:The metabolic activities are significantly altered in the serum of CMS patients.High altitude hypoxia may act on the disturbed glucose metabolism and amino acid metabolism in part of the Tibetan triggered by CMS.

OBJECTIVETo evaluate the effectiveness, safety as well as the immunological change (peripheral T cell subpopulation) in patients with idiopathic thrombocytopenic purpura (ITP) treated with lower dose rituximab.

METHODSTwenty-six patients with refractory ITP which were unresponsive to or relapse after steriod and IVIG treatment were treated with rituximab (100 mg per week for four weeks) and intravenous immunoglobulin (IVIG) treatment. Whole blood cell count, serum concentrations of IgG, IgM and IgA, platelet associated (PA)-IgG, PAIgA and PAIgM, peripheral T cell subpopulations, and B cells of CD19(+)/CD20(+) were detected before and after rituximab therapy.

RESULTSComplete response (CR) was achieved in 6 patients (23.1%), response (R) in 10 (38.5%), and non-response (NR) in 10 (38.5%). One patient relapsed after R. The median follow-up time was 5.5 (0.8 - 8) months. The median response and CR time were 27 (1 - 104) and 41 (4 - 109) days, respectively. After the therapy, the serum concentrations of IgG, IgA, IgM, T cells of CD3(+), CD3(+)CD4(+), CD3(+)CD8(+), CD3(-)CD56(+), CD4(+)CD25(+) and CD4(+)CD25(+)FOXP3(+) were not changed, the number of CD4(+)CD25(+)FOXP3(-) T cells decreased (P < 0.05) and CD19(+)CD20(+) B cells significantly decreased (P < 0.01). PAIgG was lower after treatment compared with that before treatment (P < 0.05). There were no severe adverse effects during rituximab therapy.

CONCLUSIONLower dose rituximab may be an effective and safe modality for patients with ITP.

Antibodies, Monoclonal, Murine-Derived ; therapeutic use ; B-Lymphocytes ; Humans ; Immunoglobulin G ; Purpura, Thrombocytopenic, Idiopathic ; immunology ; Rituximab

OBJECTIVESTo construct and screen a primarily phage display library of HCV C and E1 genes evolved with an artificial pattern.

METHODSTwo genes of about 1 kb with different genotypes were evolved by DNA shuffling. The re-assembled HCV C and E1 genes were cloned into a phage vector. After being rescued with helper phage M13KO7, a phage display library was constructed. Then the library was screened with anti-C and E1 McAb. Double-antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA) was carried out on twenty individual phage clones selected randomly to detect their binding and reactive activity with high-titer HCV-positive sera. Normal sera were used as controls.

RESULTSThe phage display library of HCV C and E1 genes which evolved with an artificial pattern was constructed. Their capacity amounted to 1.64 x 10(6), and 86 percent of the clones contained C and E1 genes. After four rounds of panning, the phage library was specifically enriched. Twelve positive clones were successfully screened.

CONCLUSIONThe capacity and diversity of the constructed library are enough for screening. The results demonstrate the superiority of the specific binding and reactive activity and affinity of the 12 phage clones from the HCV positive sera.

DNA, Viral ; genetics ; Gene Library ; Hepacivirus ; genetics ; Peptide Library ; Viral Core Proteins ; genetics ; Viral Envelope Proteins ; genetics

BACKGROUNDAhmed glaucoma valves (AGV) has been used for decades, but there is no detailed report about the efficacy of AGV in Chinese glaucoma patients. This study aimed to compare the intraocular pressure (IOP) lowering efficacy and side effects of S-2 polypropylene and PF-7 silicone AGV implantation in Chinese refractory glaucoma patients.

METHODSPatients were divided into S-2 model AGV group and FP-7 model AGV group. The complete and qualified surgical success rate, change of IOP, number of anti-glaucoma medications used and postoperative complications were recorded and analyzed.

RESULTSAverage follow-up time was comparable between two groups. IOP was reduced from (37.9 ± 12.7) mmHg preoperatively to (17.3 ± 5.3) mmHg at the last follow-up in S-2 group and reduced from (39.9 ± 14.4) mmHg to (17.7 ± 4.9) mmHg in FP-7 group. Anti-glaucoma medications were reduced from 3.8 ± 0.2 to 1.5 ± 0.2 in S-2 group, and 3.5 ± 0.2 to 0.7 ± 0.2 in FP-7 groups. The cumulative success rates were comparable in two groups, which were 61.2% and 72.1% in S-2 group and FP-7 group respectively. When IOP reduction criteria was used, complete success rates were 30.6% and 51.2% for S-2 and FP-7 groups, and qualified success rates were 86.1% and 92.7% separately. In both groups, the major complication was hypotony, and the previous trabeculectomy of patients was the major risk factor for surgery failure.

CONCLUSIONSIn this short-term retrospective study, S-2 AGV is showed at least as effective as FP-7 AGV in IOP reduction, but associated with higher rate of complications. Previous trabeculectomy is a principle risk factor for AGV implantation failure. These clinical outcomes are important for converting use of the FP-7 silicon AGV in Chinese refractory glaucoma patients.

Adult ; Female ; Glaucoma ; physiopathology ; therapy ; Glaucoma Drainage Implants ; Humans ; Intraocular Pressure ; physiology ; Male ; Retrospective Studies

AIMTo develop a method for assessing sperm function by measuring released acrosin activity during the acrosome reaction (AR).

METHODSHuman semen samples were obtained from 24 healthy donors with proven fertility after 3-7 days of sexual abstinence. After collection, samples were liquefied for 30 min at room temperature. Standard semen parameters were evaluated according to World Health Organization (WHO) criteria. Calcium ionophore A23187 and progesterone (P4) were used to stimulate the sperm to undergo AR. After treatment, sperm were incubated with the supravital dye Hoechst33258, fixed in a glutaraldehyde-phosphate-buffered saline solution, and the acrosomal status was determined by fluorescence microscopy with fluorescein isothiocyanate-labeled Pisum sativum agglutinin (FITC-PSA). The percentage of sperm undergoing AR (AR%) was compared to sperm acrosin activities as assessed by spectrocolorimetry. The correlation between AR% and acrosin activity was determined by statistical analysis.

RESULTSThe AR% and released acrosin activity were both markedly increased with A23187 and P4 stimulation. Sperm motility and viability were significantly higher after stimulation with P4 versus stimulation with A23187 (P < 0.001). There was a significant positive correlation between released acrosin activity and AR% determined by FITC-PSA staining (r=0.916, P < 0.001).

CONCLUSIONSpectrocolorimetric measurement of released acrosin activity might serve as a reasonable alternative method to evaluate AR.

Acrosin ; physiology ; Acrosome Reaction ; Adult ; China ; Humans ; Male ; Progesterone ; pharmacology ; Semen ; drug effects ; physiology ; Sperm Motility ; drug effects ; physiology