[Arsenic removal by coagulation process and the field expanding experiments for Yangzonghai Lake].

Yangzonghai Lake is the third largest plateau lake in Yunnan province. In June 2008, arsenic contamination was detected in Yangzonghai Lake and the water quality worsens dramatically from standard grade II to worse than grade V. Since Yongzonghai Lake is so large with the area of 31 km2 and the storage capacity of 6.04 x 10(8) m3, those pretreatment operations of the traditional arsenic removal methods, such as pre oxidation, adjusting pH value, are not applicable. In this study, a facile remediation strategy for arsenic removal by coagulation process, in which ferric chloride was directly sprayed into the contaminated water without any pretreatment, was reported. The results showed that the arsenic removal percentage was up to 95.1%-96.7% for 50 L raw water with reagent dosage of 1.62-3.20 mg x L(-1). Furthermore, the pH value of the lake kept constant in the coagulation process, which was beneficial for fish survival. Re-dissolved arsenic from precipitation was not detected in 954 days. The strategy of ferric chloride coagulation were applied to field experiments for lake water with volumes of 1 x 10(4) m3 and 25 x 10(4) m3, in which arsenic was also removed effectively. The reported strategy was of great advantage for simple operation, low cost and ecological safety, therefore it provides a representative example for arsenic contamination treatment of large lake.

Cytotoxic activity of spleen lymphocytes in BALB/c mice immunized by HSP110-HER2/neu ICD / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To explore the cytotoxic responses of spleen T lymphocytes (CTL) in BALB/c mice induced by recombinant HSP110-HER2/neu ICD complex.</p><p><b>METHODS</b>Tumor-bearing mouse model was immunized by HSP110-HER2/neu ICD complex. The IFN-γ level secreted by activated spleen T lymphocytes was detected by enzyme linked immunospot assay (ELISPOT). The corresponding CTL activity was measured by granzyme release assay.</p><p><b>RESULTS</b>The BALB/c mouse model of human mammary tumor highly expressing HER2/neu was established. HSP110-HER2/neu ICD complex immunization led to a significantly higher level of INF-γ than that in HSP110-P(789-797) immunized and HER2/neu ICD immunized mice. HSP110-HER2/neu ICD complex immunized animals also show significant CTL activity. The results of immunohistochemical staining showed that the number of blue spots in the PBS group was 4.57 ± 1.33, HSP110 group 6.83 ± 2.08, HER2/neu ICD group 16.17 ± 2.86, HSP110-P(789-797) group 43.67 ± 4.78, and SP110-HER2/neu ICD group 76.51 ± 8.17. The number of IFN-γ-secreting spleen lymphocytes in the HSP110-HER2/neu ICD group was significantly higher than that in the HSP110-P(789-797) group, and that of HSP110-P(789-797) group was significantly higher than that of HER2/neu ICD group (P < 0.01). The target cell-killing rate of the PBS group was (8.15 ± 1.27)%, HSP110 group (9.51 ± 1.51)%, HER2/neu ICD group (14.03 ± 2.45)%, HSP110-P(789-797) group (25.99 ± 3.04)% and HSP110-HER2/neu ICD group (38.15 ± 3.95)% (all P < 0.01).</p><p><b>CONCLUSIONS</b>HSP110-HER2/neu ICD complex can promote the proliferation and maturation of T lymphocytes into CTLs, and might be used as anti-tumor vaccine to induce potent cytotoxic T lymophocyte immunoresponse against specific tumor cells.</p>

Expression and purification of active recombinant human bikunin in Pichia pastoris.

Bikunin is a proteoglycan exhibiting broad-spectrum inhibitory activity against serine proteases and could potentially suppress tumor cell invasion and metastasis. Here, we have successfully expressed recombinant human bikunin (rh-bikunin) in the methylotrophic yeast Pichia pastoris and established the purification procedure. The cDNA encoding human bikunin was cloned by PCR and inserted into the expression vector pPICZalphaC. After expressed in shake flask, rh-bikunin was produced in an 80-L fermenter and purified by cation exchange chromatography and reverse phase chromatography. The rh-bikunin was active by trypsin inhibition test. The final expression levels were 55 mg/L and we got totally 1.44 g (5600 inhibitor units/mg) of purified rh-bikunin (purity is 95%) from 40 L of fermentation broth. The rh-bikunin consists of two forms with molecular masses of 24 and 21 kDa, respectively. Both forms were immunoreactive by Western blotting and N-terminals were correctly processed by amino-terminal sequencing. This study provided a new method for expression and purification of active rh-bikunin.

Reconstruction of Tissue Engineering Skin by Epidermal Cells and Fibroblasts Combined with Modified Polymer of Lactic Acid / 现代生物医学进展

Objective: To investigate the applied feasibility of scaffold with modified PLA (Polymer of lactic acid) in tissue engineering. Methods:First, we adopted salting-in method to prepare porous foam scaffold. Then, we reconstructed tissue engineering skin by epidermal cells and fibroblasts combined with modified PLA. On the 14th day of cell culturing in vitro, we was a control. Results:The arfificial skin is composed of epidermis and dermis and similar to natural skin in appearance. The skin consists of fibroblasts and keratinocytes, which are in various proliferation and differentiation stages. Fibroblasts and keratinocytes distribute on the surface of polymer of lactic acid (PLA) and the number of fibroblast and keratinocyte increase. Conclusion:Dialdehyde starches (DAS) not only improve the function of PLA but also have good effects on cells. Moreover, it does not affect the growth and the metabolism of the cells. So it is feasible to use modified scaffold to construct tissue engineering skin.

Reconstruction of Tissue Engineering Skin by Epidermal Cells and Fibroblasts Combined with Modified Polymer of Lactic Acid / 现代生物医学进展

Objective: To investigate the applied feasibility of scaffold with modified PLA (Polymer of lactic acid) in tissue engineering. Methods:First, we adopted salting-in method to prepare porous foam scaffold. Then, we reconstructed tissue engineering skin by epidermal cells and fibroblasts combined with modified PLA. On the 14th day of cell culturing in vitro, we was a control. Results:The arfificial skin is composed of epidermis and dermis and similar to natural skin in appearance. The skin consists of fibroblasts and keratinocytes, which are in various proliferation and differentiation stages. Fibroblasts and keratinocytes distribute on the surface of polymer of lactic acid (PLA) and the number of fibroblast and keratinocyte increase. Conclusion:Dialdehyde starches (DAS) not only improve the function of PLA but also have good effects on cells. Moreover, it does not affect the growth and the metabolism of the cells. So it is feasible to use modified scaffold to construct tissue engineering skin.

Photoelastic analysis of movement tendency of lower teeth using multiloop edgewise archwire with vertical elastics in anterior teeth area / 华西口腔医学杂志

<p><b>OBJECTIVE</b>To investigate the movement tendency of lower teeth using multiloop edgewise archwire (MEAW) with vertical elastics in anterior teeth area by three-dimensions photoelastic analysis.</p><p><b>METHODS</b>The photoelastical model of full teeth as human body on physical parameter and dimension was established and loaded by MEAW with vertical elastics in anterior teeth area similar with clinic. Every freezing model-tooth was bladed by anteroposterior axes and vertical axes, the stress of every point of alveolar bone was calculated by three-dimensions shear-equation method. The stress distribution regularity of per-tooth was analyzed to describe their move tendency.</p><p><b>RESULTS</b>The lower second molar was intrused and rotated to distal and inclined as negative torque. The lower first molar moved and rotated to distal, the mesial rotate to buccally and the distal rotate to lingually. The lower second pre-molar was extruded and inclined as crown to mesial and root to distal, inclined as negative torque. The lower canine was intrused slightly and inclined as positive torque. The lower lateral incisor was extruded and moved and inclined as positive torque.</p><p><b>CONCLUSION</b>Using MEAW with several "L" loop can control the movement of every tooth in three dimension.</p>

Molecular cloning and sequence analysis of ussurin, a new metalloproteinases/disintegrin from Gloydius ussuriensis / 生物工程学报

The metalloproteinases/disintegrins in the snake venom act as platelet aggregation inhibitor by an antagonism against integrin on platelet through its RGD sequence and may play other important role in cell-cell fusion, cell matrix interaction and other cellular function. Ussurin is a new metalloproteinase/disintegrin that was cloned from Gloydius ussuriensis. Poly (A+) RNA was purified from the total RNA preparation from venom gland of a single G. ussuriensis using the poly (A+) tract-mRNA isolation system. A cDNA library was constructed with the SMART PCR cDNA library construction kit. The cDNA library was screened and the positive clones were selected. The full-length cDNA of Ussurin was obtained. The cDNA encoding the Ussurin precursor has a 51bp 5'-UTR, the open reading frame of Ussurin and a 490 bp 3'-UTR, the open reading frame of Ussurin cDNA nucleotide sequence is 1434 bp and codes for 478 amino acids with a predicted molecular mass of 53.2 kD and an isoelectric point of 5.37. There is no potential N-glycosylation site in the deduced sequence region. Its deduced amino acid sequence consists of four region, a signal sequence of 18 amino acid residues, a zymogen pro-peptide of 171 amino acid residues with a cysteine switch motif (PK-MCGVT) in it, a central metalloproteinase domain of 201 amino acid residues containing a conserved zinc-chelating sequence (HEXXHXXGXXH) and a methionine-turn CIM involving zinc banding also, a space sequence between metalloproteinase domain and disintegrin domain of 15 amino acid residues with a conserved T392, T397, S400, which is specific residues of the P-II snake venom metalloproteinases, a disintegrin domain of 73 amino acid residues with a characteristic RGD region and six-disulfide bonds. Ussurin belongs to P-II class. The cDNA sequence and deduced amino acid sequence of Ussurin precursor were compared with homologous sequence in the GenBank database, the result reveals high degree of homology in sequence and organization pattern of domain with metalloproteinase/disintegrin gene family of other snake species. Compared with the alignment of amino acid sequence of metalloproteinase/disintegrin member, hypervariable regions of this member were revealed, besides they share higher homologous in the zymogen domain. It suggests that the hypervariable regions are the counterparts directly suitable for interacting with different domain of receptors, different receptors or substrates.