[Temporal-spatial Variation and Source Identification of Hydro-chemical Characteristics in Shima River Catchment, Dongguan City].

Shima River catchment is of strategic importance to urban water supply in Dongjiang portable water source area. To investigate the hydro-chemical characteristics of Shima River, 39 river water samples were collected in February, June and November, 2012 to analyze the major ions (K+, Na+, Ca2+, Mg2+, Cl-, SO4(2-) , HCO3-) and nutritive salts (PO4(3-), NO3- and NH4+) and to discuss the temporal-spatial variation and controlling factors of hydro-chemical composition, relative sources identification of varied ions was performed as well. The results showed that the hydro-chemical composition exhibited significant differences in different periods. The average concentration of total dissolved solid ( TDS) and nutritive salts in different investigated periods followed the decreasing order of November > February > June. The dominant anion of Shima River was HCO3-, and Na+ + K+ were the major cations in February and November which were changed to Ca2+ in June, the hydro-chemical types were determined as HCO(3-)-Na+ and HCO(3-)- Ca2+ in dry (February and November) and rainy (June) seasons, respectively. Spatial variations of concentration of nutritive salts were mainly affected by the discharges of N- and P-containing waste water resulted from human activities. The ratio between N and P of water sample (R7) was 18.4:1 which boosted the "crazy growth" of phytoplankton and led to severe eutrophication. According to Gibbs distribution of water samples, dissolution of hydatogenic rocks was the primary factor to control the major cations of river water in dry season, however, the hydro-chemical composition was significantly affected by the combination of hydatogenic and carbonate rocks in rainy season. The deposition of sea-salts contributed less to chemical substances in river. Correlation analysis revealed that K+, Na+, Mg2+, Cl- and SO4(2-) were partly derived from the application of fertilizer and the discharge of industrial effluent; Waste water of poultry feeding and sanitary wastewater transported large quantities of NH(4+)-N, PO4(3-)-P and NO(3-)-N into the river.

Intranasal immunization with chitosan/pCAGGS-flaA nanoparticles inhibits Campylobacter jejuni in a White Leghorn model.

Campylobacter jejuni is the most common zoonotic bacterium associated with human diarrhea, and chickens are considered to be one of the most important sources for human infection, with no effective prophylactic treatment available. We describe here a prophylactic strategy using chitosan-DNA intranasal immunization to induce specific immune responses. The chitosan used for intranasal administration is a natural mucus absorption enhancer, which results in transgenic DNA expression in chicken nasopharynx. Chickens immunized with chitosan-DNA nanoparticles, which carried a gene for the major structural protein FlaA, produced significantly increased levels of serum anti-Campylobacter jejuni IgG and intestinal mucosal antibody (IgA), compared to those treated with chitosan-DNA (pCAGGS). Chitosan-pCAGGS-flaA intranasal immunization induced reductions of bacterial expellation by 2-3 log(10) and 2 log(10) in large intestine and cecum of chickens, respectively, when administered with the isolated C. jejuni strain. This study demonstrated that intranasal delivery of chitosan-DNA vaccine successfully induced effective immune response and might be a promising vaccine candidate against C. jejuni infection.

Microglial activation and age-related dopaminergic neurodegeneration in MPTP-treated SAMP8 mice.

Senescence-accelerated mouse prone 8 (SAMP8) has an early onset of senility and a shorter life span, providing with cognitive impairment. Contrasted with C57BL/6 mouse, which is most commonly used in the study of Parkinson's disease (PD), SAMP8 needs shorter period of breeding and might be good candidate for the investigation of cognitive impairment in PD. Studies had shown the increase of sensibility to 1-methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridine (MPTP) with aging in C57BL/6 mouse. However, the sensitivity of MPTP neurotoxicity depends on the strains of animal and the exact mechanisms of the progression of PD promoted by aging is lack of consensus. Here, we showed after MPTP injection, the spontaneous activity of both young (3-month-old) and old (6-month-old) SAMP8 decreased dramatically, and the old mice required longer recovery time. Immunohistochemical and immunoblot analysis revealed that old mice displayed significant reductions in the dopaminergic neuron numbers and tyrosine hydroxylase (TH) protein. Microglia protein (CD11b) in the striatum of old mice increased more pronouncedly than that in the young mice from 24 h to 3 days. Inducible nitric oxide synthase (iNOS) in the striatum remarkably increased, however, no discernible difference between the two groups was found. These results suggested that the sensibility to MPTP increased with aging in SAMP8. A greater increase of microglial activation in old mice may be a possible mechanism to explain how advancing age predisposes the dopamine system to parkinsonism. The MPTP-SAMP8 model will start a new consideration for the study of PD.

[Preparation and property of platinum microcoil modified by a copolymer-VEGF conjugate].

OBJECTIVE: To prepare a platinum microcoil coated with polymers and vascular endothelial growth factor (VEGF), and evaluate its surface characteristics and property of sustained VEGF release. METHODS: The surface of the platinum microcoils (GDC) were modified by coating P(DLLA-co-TMC) copolymer and immobilizing heparin on the surface of GDC. VEGF was then loaded onto the surface of GDC and the controlled release of VEGF within GDC was achieved. The morphology was observed by scanning electron microscope, and the sustained release of VEGF was evaluated by enzyme-linked immunosorbent assay (ELISA). RESULTS: Platinum coils were prepared by successive deposition of P(DLLA-co-TMC) copolymer and anionic heparin, and VEGF was immobilized through affinity interaction with heparin. The accumulative release of VEGF increased obviously during the entire testing period without burst release. CONCLUSION: The use of P(DLLA-co-TMC) copolymer allows immobilization of VEGF on the platinum coils for controlled VEGF release, and improves the biological property of the coils.

Neuroprotective effects of enriched environment in MPTP-treated SAMP8 mice.

Neuroprotective effects of enriched environment (EE) have been well established. Recent study suggests that exposure to EE can protect dopaminergic neurons against MPTP-induced Parkinsonism. After 64 female SAMP8 mice were reared in EE and standard environment (SE) for 3 months, the effects of EE and SE were compared on behavioural change, tyrosine hydroxylase (TH) immunoreaction positive neuron and dopaminetransporter (DAT) expression in 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine(MPTP)-treated SAMP8. EE mice showed decreased spontaneous activity compared with SE mice. But EE+MPTP mice showed less decreased spontaneous activity compared with SE+MPTP mice. Otherwise, EE mice showed increased percentage of entries into the open arms and percentage of time spent in the open arms. Furthermore, EE mice demonstrated reduced neurotoxicity, with less decreased TH mRNA and protein expression in Substantia Nigra (SN) after MPTP administration compared with SE mice. SE mice showed a 53.77% loss of TH-positive neurons, whereas EE mice only showed a 42.28% loss. Moreover, EE mice showed decreased DAT mRNA and protein expression compared with SE mice. These data demonstrate that EE can protect dopaminergic neurons against MPTP-induced neuronal damage, which suggest that the probability of developing Parkinson's disease (PD) may be related to life environment.

Regulation of vascular smooth muscle cells on poly(ethylene terephthalate) film by O-carboxymethylchitosan surface immobilization.

Specifying the chemical environment of cells is a well-established method of controlling cellular behaviors. In this study, poly(ethylene terephthalate) (PET) film was selected as a typical biomaterial to detect the effects of chemical modifications on material surface in controlling cell behaviors. Natural biopolymer chitosan and its biocompatible derivative, O-carboxymethylchitosan (OCMCS) were surface immobilized on PET, respectively, via argon plasma followed by graft copolymerization with acrylic acid (AAc), which was exploited to covalently couple PET with chitosan (CS) and OCMCS molecules. Smooth muscle cells (SMCs) displayed a surface-dependent cell spreading and cytoskeletal organization. The cells spread with a more pronounced elongated spindle shape, smaller cell area, and lower cell shape index (CSI) on OCMCS-modified PET surface than on PET, or the PAA and chitosan-immobilized PET surfaces after 24 h of culture. Cell-culture viability after 5 days showed that all the modified materials possessed good cell proliferation. Our results suggest that cell adhesion, morphology, and growth can be mediated not only by varying the functional groups, electric charge, and wettability of PET surface but also by the specific biological recognition elicited from the biomaterials. These findings strongly support the concept that the microenvironment significantly influences cell behavior, highlighting the importance of environmental material biochemistry in cell-based tissue engineering schemes.

Trace measurement of phenothiazine drugs in tablets by micellar-enhanced fluorophotometric method.

It was found that in buffer solution of pH 7.0, the addition of sodium dodecyl sulfate (SDS) to the solution of phenothiazine drugs, such as chlorpromazine, promethazine and trifluoperazine, showed a remarkable enhancement of their fluorescence intensity. A further study proved that the phenothiazine drugs can be determined by fluorophotometric method in micellar system. Under optimal conditions, there was a good linear relationship between fluorescence intensity and phenothiazine compounds concentration, and the detection limit of 3.0 x 10(-8) M chlorpromazine, 3.0 x 10(-8) M promethazine and 1.5 x 10(-8) M trifluoperazine (S/N=3) were also obtained. This method has been used to determine phenothiazine drugs in tablets with satisfactory results.

Three-dimensional microchannels in biodegradable polymeric films for control orientation and phenotype of vascular smooth muscle cells.

The poor mechanical strength and vasoactivity of current small-diameter tissue engineered blood vessels (TEBVs) remain unsolved problems. Given the plasticity of smooth muscle cells (SMCs), 1 of the main limitations of current scaffolding techniques is the difficulty in controlling SMC phenotype shifts in vitro. A synthetic phenotype allows the cells to rapidly proliferate and produce extracellular matrix (ECM), whereas a shift to contractile phenotype with organized ECM ultimately provides a functional blood vessel. In this study, 3D deep (65 microm) and wide microchannels separated by high-aspect ratio (8) microwalls were successfully ultraviolet (UV) microembossed using a liquid UV polymerizable biodegradable macromer (poly(epsilon-caprolactone-r-L-lactide-r-glycolide) diacrylate) and the in vitro guidance effects of varying channel width (40-160 microm) on SMCs were verified. The results show that SMCs cultured in the wider microchannels (80-160 microm wide) switch from fibroblast morphology and random orientation to spindle-shaped morphology, and align along the direction of the microchannel nearing confluence achieved with similar cell density to unpatterned film. Further, an enhanced expression of smooth muscle alpha-actin of SMCs grown on micropatterns was found nearing confluence, which demonstrates a phenotype shift to a more contractile phenotype. These films are flexible and can be folded into tubular and lamellar structures for tissue engineering of small-diameter TEBVs as well as other organs such as esophagus or intestine. These results suggest that these micropatterned synthetic biodegradable scaffolds may be useful for guiding SMCs to grow into functional, small-diameter vascular grafts.

Foldable micropatterned hydrogel film made from biocompatible PCL-b-PEG-b-PCL diacrylate by UV embossing.

Foldable hydrogel films with micropatterns measuring 480 microm by 45 microm by 54 microm by 2 cm (width of microchannel by width of microwall by height of wall by length of pattern) were made by UV embossing of a block copolymer of polycaprolactone (PCL) and poly(ethylene glycol) (PEG), specifically PCL-b-PEG-b-PCL-diacrylate (DA), with a polydimethylsiloxane mold. The mold was treated with Ar/CF(4) plasma to simultaneously promote microchannel filling and demolding, and the glass substrate was modified with 3-(trimethoxysilyl) propyl acrylate to promote hydrogel adhesion to avoid delamination of the gel during demolding. The micropatterned hydrogel film was detached from the glass substrate by freeze-drying. As the films were demolded, the microstructured pattern was well replicated in the hydrogel. The gel pattern dimensions shrank with freeze-drying and increased with water swelling, but under both conditions, the gel micropattern morphology was perfectly preserved. PCL-b-PEG-b-PCL-DA hydrogel was found to have good biocompatibility compared with PEGDA hydrogel. A micropattern with a smaller microchannel width of 50 microm was also made. Micropatterned foldable and biocompatible hydrogel films have potential applications in the construction of tissue-engineering scaffolds.

Adhesion contact dynamics of 3T3 fibroblasts on poly (lactide-co-glycolide acid) surface modified by photochemical immobilization of biomacromolecules.

A simple and effective method of biomacromolecule immobilization on biomaterial surface for direct tuning of biophysical parameters such as the initial cell deformation rate, degree of cell spreading and adhesion kinetics is important for tissue engineering. The photochemical immobilization of azide-chitosan (Az-CS) on poly (lactide-co-glycolide) acid (PLGA) is applied here. Chitosan immobilization on PLGA through the photoactive azide group further facilitates subsequent grafting of other biocompatible biomacromolecules like gelatin (Gel) through the active amine groups on CS. This study quantitatively compares the 3T3 fibroblast adhesion dynamics on three PLGA surfaces (Gel-CS-PLGA, CS-PLGA and unmodified PLGA surfaces) using Confocal-Reflectance Interference Contrast Microscopy (C-RICM) together with phase contrast imaging. CS-PLGA and Gel-CS-PLGA surfaces developed were confirmed by X-ray photoelectron spectroscopy, atomic force microscopy and water contact angle and cell adhesion contact dynamics measurements. The cell adhesion was strongest on the Gel-CS-PLGA surface and lowest on unmodified PLGA. The steady state adhesion energy attained by the cells on gelatin modified PLGA surface is determined as 4.0 x 10(-8) J/m(2), which is about 400 times higher than that on PLGA surface (1.1 x 10(-10) J/m(2)). Significantly increased cell adhesion with Gel-CS-PLGA is postulated to result in increased cell spreading. Our integrated biophysical method can quantify the transient contact dynamics and is sufficiently accurate to discriminate even between Gel and CS modified surfaces.

Adhesion dynamics, morphology, and organization of 3T3 fibroblast on chitosan and its derivative: the effect of O-carboxymethylation.

Chitosan and O-carboxymethylchitosan (OCMCS) have been proved to have biocompatibility and have been extensively researched in the field of biomaterials. In this study, Confocal-reflectance interference contrast microscopy (C-RICM) in conjunction with phase contrast imaging was used to investigate the adhesion contact dynamics of 3T3 fibroblasts on chitoan and OCMCS surface-modified silica coverslips. The C-RICM results demonstrate that the weak cell contact forms on OCMCS surface while a much stronger contact area forms on the chitosan surface. 3T3 fibroblasts are found to spread randomly with spindlelike morphology on the chitosan surface, while they exhibit elongated morphology and align on the OCMCS surface. It is believed that fibroblast behaviors such as migration, spreading with an elongated morphology, and alignment on the OCMCS surface are correlated with the weak cell contact. The mechanisms to form cell adhesion contact on chitosan and OCMCS were discussed.

Interaction between O-carboxymethylchitosan and dipalmitoyl-sn-glycero-3-phosphocholine bilayer.

O-carboxylmethylchitosan (OCMCS), a chitosan derivative, has emerged as a strong polymeric biomembrane perturbant. In this study, the interaction between OCMCS and dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) was examined with cross-polarization microscopy, differential scanning calorimetry (DSC) and the surface pressure-area isotherms techniques. Cross-polarized light images showed that OCMCS induced the fusion of small DPPC multilamellar vesicles (MLV) to form large lamellar structures. From DSC measurement, the highest degree of fusion was found at the optimum OCMCS concentrations between 0.0625 and 0.2 mg/ml which are orders of magnitude lower than those required for similar reductions with unmodified chitosan as perturbant. At these concentrations, the association of DPPC and OCMCS enhances the fusion of DPPC vesicles. Surface pressure-area isotherms of DPPC monolayer in the presence of OCMCS imply interactions between OCMCS and DPPC in neutral condition. In comparison with chitosan, OCMCS proved to be a more effective membrane perturbant not only in neutral but also in acidic and basic conditions. The physical driving forces for OCMCS-induced perturbation of DPPC bilayer in neutral conditions are mainly hydrogen bonding and hydrophobic interactions. In acidic or basic conditions, the physical driving forces are dominated by the electrostatic interactions. The strong OCMCS-DPPC interaction will potentially increase the effectiveness of OCMCS for gene or drug delivery.

Cell viability of chitosan-containing semi-interpenetrated hydrogels based on PCL-PEG-PCL diacrylate macromer.

Chitosan-modified biodegradable hydrogels were prepared by UV irradiation of solutions in mild aqueous acidic media of poly(caprolactone)-co-poly(ethylene glycol)-co-poly(caprolactone) diacrylate (PCL-PEG-PCL-DA) and chitosan. Hydrogels obtained were characterized using FT-IR, DSC, TGA and XPS. FT-IR, TGA and DSC revealed the semi-interpenetrating polymer network structure formed in the hydrogel. Though the water swelling degree of these chitosan-modified hydrogels was substantial in the range of 322-539%, it was found that fibroblasts could still attach, spread and grow on them; this is in contrast to the commonly investigated PEG-diacrylate hydrogel. The MTT assay demonstrated that cells could grow better on 3 or 6% chitosan-modified hydrogel than unmodified PCL-PEG-PCL-DA hydrogels or low-content (1%) chitosan-modified PCL-PEG-PCL-DA hydrogel. Increased chitosan content resulted in increased cell interaction and also decreased water swelling, both of which results in increased cell attachment and spread.

[Genetic polymorphism of six short tandem repeat loci in the Han population in Hebei province of China].

OBJECTIVE: This article reports a population genetic study on six short tandem repeat(STR) loci, D7S820, D19S253, D12S391, D5S818, D16S539 and D8S1179, in a sample of unrelated Chinese Han individuals(n=122-173) living in Hebei province. METHODS: DNA extraction from blood samples (200 in number) and multiplex amplification of the above six loci were carried out. Using denaturing polyacrylamide gel electrophoresis and silver stain, the authors investigated the distribution of allele frequencies of the six loci in Han population in Hebei province. RESULTS: The STR polymorphisms at all of the six loci were observed in Chinese Han population in Hebei province. The observed heterozygosities of D7S820, D19S253, D12S391, D5S818, D16S539 and D8S1179 were 0.828, 0.757, 0.769, 0.837, 0.785 and 0.852, respectively. The measured values of the power of discrimination (PD) were 0.914, 0.919, 0.940, 0.909, 0.917, 0.944; of the mean exclusion chance(MEC) 0.618, 0.740, 0.801, 0.557, 0.655, 0.696 and of the polymorphism information content (PIC) in Chinese 0.771, 0.760, 0.762, 0.708, 0.776 and 0.794, respectively. CONCLUSION: The genotype distributions of the six STR were in accordance with Hardy-Weinberg equilibrium. The numerical values of the PD and MEC are relatively high in Hebei province, and thus can be of significant application in population genetics and forensic medicine.

[Attachment and growth of cultured fibroblast cells on chitosan/PHEA-blended hydrogels].

The chitosan/PHEA-blended hydrogels were prepared from PHEA and chitosan in various blend ratios. The water contents of the hydrogels were in the range of 50%-80% (wt). The attachment and growth of fibroblast cells(L929) on the hydrogels were studied. The results indicated the PHEA content in hydrogels has great effect on cell attachment but has little effect on the growth of L929 cells.