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W J Li

Publications and source records attributed to W J Li.

At least 19 recordsLinked to original sources

Polymer/alginate amalgam for cartilage-tissue engineering.

Marrow stroma-derived cells (MSC) are highly proliferative, multipotential cells that have been considered as ideal candidate cells for autologous tissue engineering applications. In this study, we have characterized the chondrogenic potential of human MSCs in both a PLA/alginate amalgam and pure PLA macrostructure as model three-dimensional constructs to support both chondrogenic differentiation and proliferation following TGF-beta treatment. MSCs were seeded in experimental groups that consisted of PLA-loaded constructs and PLA/alginate amalgams with and without recombinant human TGF-beta1. Chondrogenesis of the PLA and the PLA/alginate amalgam cultures was assessed at weekly intervals by histology, immunohistochemistry, scanning electron microscopy, sulfate incorporation, and RT-PCR. Chondrogenic differentiation occurs within a polymeric macrostructure with TGF-beta1 treatment as indicated by histological, immunohistochemical, sulfate incorporation, and gene expression profiles. This macrostructure can be further encased in an alginate gel/solution to optimize cell shape and to confine growth factors and cells within the polymer construct, while the polymeric scaffold provides appropriate mechanical/tissue support. The stable three-dimensional PLA/alginate amalgam represents a novel candidate system of mesenchymal chondrogenesis, which is amendable to investigation of mechanical and biological factors that normally modulate cartilage development and formation as well as a potential tissue engineering construct for cartilage repair.

Alginates↗

Three-dimensional cartilage formation by bone marrow-derived cells seeded in polylactide/alginate amalgam.

Bone marrow-derived cells are considered as candidate cells for cartilage tissue engineering by virtue of their ability to undergo chondrogenesis in vitro when cultured in high density or when embedded within a three-dimensional matrix in the presence of growth factors. This study evaluated the potential of human bone marrow-derived cells for cartilage tissue engineering by examining their chondrogenic properties within a three-dimensional amalgam scaffold consisting of the biodegradable polymer, poly-L-lactic acid (PLA) alone, and with the polysaccharide gel, alginate. Cells were suspended either in alginate or medium and loaded into porous PLA blocks. Alginate was used to improve cell loading and retention within the construct, whereas the PLA polymeric scaffold provided appropriate mechanical support and stability to the composite culture. Cells seeded in the PLA/alginate amalgams and the plain PLA constructs were treated with different concentrations of recombinant human transforming growth factor-beta1 (TGF-beta 1) either continuously (10 ng/mL) or only for the initial 3 days of culture (50 ng/mL). Chondrogenesis was assessed at weekly intervals with cultures maintained for up to 3 weeks. Histological and immunohistochemical analysis of the TGF-beta 1-treated PLA/alginate amalgam and PLA constructs showed development of a cartilaginous phenotype from day 7 to day 21 as demonstrated by colocalization of Alcian blue staining with collagen type II and cartilage proteoglycan link protein. Expression of cartilage specific genes, including collagen types II and IX, and aggrecan, was detected in TGF-beta 1-treated cultures by reverse transcription-polymerase chain reaction analysis. The initiation and progression of chondrogenic differentiation within the polymeric macrostructure occurred with both continuous and the initial 3-day TGF-beta 1 treatment regimens, suggesting that key regulatory events of chondrogenesis take place during the early period of cell growth and proliferation. Scanning electron microscopy revealed abundant cells with a rounded morphology in the PLA/alginate amalgam. These findings suggest that the three-dimensional PLA/alginate amalgam is a potential candidate bioactive scaffold for cartilage tissue engineering applications.

Alginates↗

Streptimonospora salina gen. nov., sp. nov., a new member of the family Nocardiopsaceae.

Actinomycete strain YIM 90002T (= CCTCC 99003T = CCRC 16284T) was isolated from a soil sample collected from a salt lake in the west of China. The aerial mycelium of this organism is well developed but not fragmented and, at maturity, forms short chains of spores. Spores in short chains are oval- to rod-shaped and have wrinkled surfaces. Substrate mycelium is branched with non-fragmenting hyphae and forms single oval to round spores borne on sporophores or dichotomously branching sporophores. Single spores have wrinkled surfaces. Single spores and spores in short chains are non-motile. Strain YIM 90002T contains meso-diaminopimelic acid, DD-diaminopimelic acid, glycine, lysine and aspartic acid in its cell wall and has glucose, galactose, ribose, xylose, arabinose and mannose as whole-cell sugars (no diagnostic sugars). The phospholipids are phosphatidylglycerol, phosphatidylinositol and phosphatidylethanolamine. The major menaquinones are MK-9(H6), MK-10(H2) and MK-10(H4). Phylogenetic data indicate that this strain belongs to the family Nocardiopsaceae. The morphological and physiological characteristics and chemotaxonomic and phylogenetic data for this strain differ from those of previously described actinomycetes. Therefore, a new genus, Streptimonospora, is proposed for this organism; the type species of the genus is Streptimonospora salina gen. nov., sp. nov., and the type strain of S. salina is strain YIM 90002T.

Actinomycetales↗

Frequent inactivation of the TP53 gene in esophageal squamous cell carcinoma from a high-risk population in China.

Esophageal squamous cell carcinoma (ESCC) is one of the most common fatal cancers worldwide, and north central China has some of the highest rates in the world. Previous studies from tumors in this area of China have shown high frequencies of allelic loss on chromosome 17p13-11, which includes the region where the TP53 gene is found. We examined 56 ESCC patients using single-strand conformation polymorphism and DNA sequencing to assess the frequency and spectrum of TP53 mutation and the association between allelic loss at microsatellite marker TP53 and TP53 mutations. Ninety-six % of cases were found to have at least one genetic alteration, including TP53 mutation (77%), allelic loss within the TP53 gene (73%), and/or loss of heterozygosity at the TP53 microsatellite marker (80%); 75% had two or more such alterations, including 59% with both a point mutation and an intragenic allelic loss ("two hits"). The majority of mutations observed were in exon 5, where the most common type of nucleotide substitution was a G:C-->A:T or C:G-->T:A transition, including half that occurred at CpG sites. Allelic loss was most commonly found in exon 4 but was very common in exon 5 as well. Taken together, the multiple genetic alterations of TP53 in this population at high risk for ESCC indicate that there is a very high degree of genetic instability in these tumors, that TP53 is a primary target for inactivation, and that this tumor suppressor gene plays a critical role in the carcinogenesis process for ESCC.

Adult↗

Estimation of the radionuclide distribution in sediment in coast area.

The study of the radionuclide distribution in sediment is a very important aspect in environmental impact of the low level radioactive liquid waste (LLW) from coastal nuclear facilities or nuclear power plant. Even now we do not know much about it. In this paper, a simple and useful method is put forward and it is used to estimate the nuclide distribution in sediment. The result showed that the LLW from nuclear facility or nuclear power plant will do a little harm to the sediment nearby. But the harm is not very serious. Much works have to be done before full understanding of the situation.

Environmental Monitoring↗

Molecular cloning and expression of a phospholipid hydroperoxide glutathione peroxidase homolog in Oryza sativa.

A cDNA encoding putative phospholipid hydroperoxide glutathione peroxidase (PHGPX) was isolated from rice using rapid amplification of cDNA ends. This cDNA, designated ricPHGPX, includes an open reading frame encoding a protein of 169 amino acids which shares about 60% and 50% amino acid sequence identity with plant and mammalian PHGPXs, respectively. The gene is expressed at a relative high level in flag leaves and the expression can be markedly induced by oxidative stress, suggesting that the product of the gene plays a key role in defense against oxidative damage in rice.

Amino Acid Sequence↗

[Simultaneous analysis of low concentrations of glucose, ethanol and glycerol by high performance liquid chromatographic method].

A method based on high performance liquid chromatography (HPLC) for the analysis of glucose, ethanol and glycerol simultaneously is presented. The baseline separation was achieved at room temperature (25-30 degrees C) on a Beckman mu-Spherogel carbohydrate column (6.5 mm i.d. x 30 cm) with H2SO4-H2O(0.5:1,000, V/V) as mobile phase at a flow rate of 1.000 mL/min. All the substances were detected with the Beckman 186 Refractive Index Detector. Under these conditions, distinct peaks of glucose, ethanol and glycerol were resolved within 15 min. The retention time in the particular condition was (6.04 +/- 0.04) min for glucose, (13.36 +/- 0.08) min for ethanol and (8.72 +/- 0.05) min for glycerol. The detection limit for glucose, ethanol and glycerol were found to be 10(-5) g, 10(-4) g and 10(-5) g respectively. A series of experiments have been performed to investigate the glucose consumption and the ethanol, glycerol production in the random growth of Saccharomyces cerevisiae with the method above. Only 0.5 mL sample was needed each time. In 30 hours after inoculation, the cell density increased from 10(6) mL-1 to 10(7) mL-1. It was found that the glucose concentration decreased approximately linearly after 8 hours, and ethanol and glycerol was detected after the delay period as expected. The results show that the method proposed is rapid and convenient with enough accuracy. The method may be used to investigate the metabolic behavior of yeast in the fermentation process at the early stage or detect the slight changes of concentrations of some objective substances in particular culture experiments.

Chromatography, High Pressure Liquid↗

Alleviation of pre-exposure to low-dose 16O8+ ion on mouse testicular histological damage induced by subsequent high-dose irradiation.

The testes of the B6C3F1 hybrid strain mice were irradiated with 0.05 Gy of 16O8+ ion as the pre-exposure dose (D1), and were then irradiated with 2 Gy of 16O8+ ion as challenging radiation dose (D2) at 4 h after per-exposure. Testicular morphology was observed by light microscope at 35th day after radiation. The results showed that irradiation of mouse testes with 2 Gy of 16O8+ ion significantly impaired, mainly reduction of tubule diameter and decrease or loss of germ cells in various developing stages, especially spermatogenic elements. Pre-exposure to a low-dose (0.05 Gy) of 16O8+ ion significantly alleviated above mentioned damage on testicular morphology induced by subsequent a high-dose (2 Gy) radiation.

Animals↗

Effects of 16O+6 ion irradiation on human sperm spontaneous chemiluminescence, motility, acrosome reaction and viability in vitro.

Effects of 16O+6 ion irradiation with different doses on human sperm spontaneous chemiluminescence (SCL), motility, acrosome reaction (AR) and viability were examined. Spermatozoa were irradiated with 0, 0.25, 0.5, 1, 2, 4, 8, 16, 32, or 64 Gy 16O+6 ion beam at the energy of 3.17 MeV/u. After irradiation, samples were analyzed by SCL measurement at 1, 2 and 3 h of incubation; motility was determined by the transmembrane migration method within 2 h of incubation; the percentage of AR and viability was evaluated by the triple-stain technique at 3.5 h of incubation. The results showed: sperm SCL was significantly increased with irradiation doses and the lowest effective dose was 0.5 Gy; compared with controls, the transmembrane migration ratio of spermatozoa progressively elevated with irradiation doses at 0.5, 1, and 2 Gy; the percentage of sperm AR markedly increased in 0.5-4 Gy irradiation and the optimal dose was 2 Gy, and then significant decreased with further increase of irradiation doses; the viability had no significant change within 0.25-8 Gy, but was progressively decreased at 16, 32 and 64 Gy. These data suggested that heavy ion at low doses increased motility and AR, whereas had deleterious effects at higher doses, which are associated with free radical reactions induced by heavy ion irradiation.

Acrosome Reaction↗

Chromosomal aberrations induced by 12C6+ heavy ion irradiation in spermatogonia and spermatocytes of mice.

The testes of Kun-Ming strain mice were radiated with different doses of 12C6+ ion or 60Co gamma-ray. Chromosomal aberrations induced in spermatogonia and spermatocytes were analyzed by the air-drying method. The relative biological effectiveness (RBE) of 12C6+ ion was calculated with respect to 60Co gamma-ray for the induction of chromosomal aberrations. The 12C6+ ion and 60Co gamma-ray dose-response relationships for chromosomal aberrations were plotted by linear quadratic models. The results showed that there was an increase in frequency of chromosomal aberrations in all the treated groups compared to controls. The RBE values were 1.67 for aberrations of spermatogonia and 1.66 for aberrations of spermatocytes for a dose of 2.0 Gy. Moreover, a different distribution of the various types of aberrations has been found for 12C6+ ion and 60Co gamma-ray irradiations. The dose-response relationships for 12C6+ ion and 60Co gamma-ray exhibited negative curvature in both spermatogonia and spermatocytes groups: the frequencies of aberrations increased sharply at low doses and exhibited less sharp increases for higher doses, which may be related to an interaction between the chromosomal damage and a block in cell cycle. Our results may provide useful information for the assessment of genetic risks of humans exposed to heavy ions.

Animals↗

Effects of pre-exposure of mouse testis with low-dose (16)O8+ ions or 60Co gamma-rays on sperm shape abnormalities, lipid peroxidation and superoxide dismutase (SOD) activity induced by subsequent high-dose irradiation.

PURPOSE: To investigate the effects of pre-exposure of mouse testis with low-doses of (16)O8+ ions or 60Co gamma-rays on sperm shape abnormalities, lipid peroxidation and superoxide dismutase (SOD) activity induced by subsequent high-dose irradiation. MATERIALS AND METHODS: Testes of the B6C3F1 hybrid strain mice were pre-irradiated with 0.05 Gy of (16)O8+ ions or 60Co gamma-rays and then after 4 h given a test irradiation with 2 Gy of the same radiation type. SOD activity and thiobarbituric acid reactive substances (TBARS) in the testes were determined by spectrophotometric and TBA methods respectively at 4 h after irradiation. Testis weight, sperm count and sperm morphology were analysed at day 35 after irradiation. RESULTS: Compared with controls, there was a significant increase in SOD activity and a significant decrease in TBARS level of pretreated testes. Testis weight loss, sperm count reduction and sperm abnormalities were significantly lower in the pretreated testes. The bioeffects of a 2 Gy dose of (16)O8+ ions relative to 60Co gamma-rays were 1.84 +/- 0.28 for testis weight, 1.22 +/- 0.25 for sperm count and 1.29 +/- 0.10 for sperm abnormalities. CONCLUSIONS: These data suggest that pre-exposure of testes with a low dose of heavy ions or gamma-rays renders the organ more resistant to subsequent high-dose irradiation. The increase of SOD activity and the decrease of lipid peroxidation levels induced by low-dose ionizing irradiation may be involved in this resistance. The effects with heavy ion irradiation were greater than with gamma-rays.

Animals↗

The production and preclinical characterization of a chimeric anti-breast-cancer antibody, cBC2.

A chimeric (mouse-human) BC2 antibody (cBC2) was produced which may be used in the diagnosis and treatment of breast cancer. The BC2 variable region genes were amplified by polymerase chain reaction (PCR), using oligonucleotide primers homologous to the framework sequences of mouse VH and V kappa genes. The PCR products were used to create cBC2 expression vectors containing the mouse BC2 VH and V kappa and human constant region (IgG1 and K) genes. Chimeric antibody was produced following transfection of these constructs into Sp2/0 myeloma cells. Binding assays in vitro demonstrated that cBC2 had the same specificity for human milk fat globule membrane (HMFGM) and MUC1+ cells as mBC2, and bound antigen with a similar affinity (cBC2, Ka 5.53 +/- 2.09 x 10(8); mBC2, Ka 1.44 +/- 0.98 x 10(9)). Functionally, only cBC2 (5-25 micrograms ml-1), was able to mediate antibody-dependent cellular cytotoxicity (ADCC) with human effector cells, with 25% maximal specific lysis of MUC1+ cells at an E/T ratio of 100:1. Human complement-mediated lysis was minimal (10-15% specific lysis) with both mBC2 and cBC2. Neither cBC2 nor mBC2 was able to inhibit tumour growth in vivo in the absence of covalently coupled anticancer drugs. However, biodistribution studies demonstrated that both antibodies preferentially targeted MUC1+ tumour cells, with 17% of the injected dose of cBC2, as compared to 27% of mBC2, localized to the MUC1+ tumour at 24 h (less than 6% detected in any other tissue).

3T3 Cells↗

[Medullipin].

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Animals↗

[Target therapy by monoclonal antibody against ovarian carcinoma conjugated with liposomes and adriamycin].

Conjugates of monoclonal antibody COC166-9 and adriamycin entrapped in liposome (MLA) were prepared in our laboratory. In vitro growth inhibition of SKOV3 ovarian carcinoma cell line was carried out by MLA and controls. Target therapies by MLA, adriamycin, normal mouse IgG instead of MAb, and control were given for 24 nude mice models with subcutaneous human ovarian carcinoma xenografts and 16 ascitic ovarian carcinoma respectively. MLA group showed the best therapeutic effect than all the other groups which gave a helpful clue to clinical use.

Animals↗

[Target therapy by monoclonal antibody against ovarian carcinoma conjugated with liposome and adriamycin].

Conjugates of monoclonal antibody (MAb) and adriamycin entrapped in liposome (MLA) were prepared by COC166-9. The MAb against ovarian serous adenocarcinoma was generated in our laboratory. Target therapies were done in nude mice model with subcutaneous tumor xenografts in 24 and ascitic carcinoma in 16 by MLA. The results demonstrated that MLA group presented the best therapeutic effect than all the other groups, which gave a very helpful clue to clinical target therapy of ovarian carcinoma in the future.

Animals↗

[The induction of specific immunity to human ovarian carcinoma cells by anti-idiotypic antibodies].

Monoclonal anti-idiotypic antibodies, 6B11 and H12, were employed instead of the tumor antigen to induce a specific cellular immunity to ovarian carcinoma cell line SKOV3. 6B11 and 1H12 carring the internal image of ovarian carcinoma antigen. BALB/c mice were immunized with 6B11 or 1H12 and then the mouse foot pad was attacked by the injection of target cell SKOV3 to stimulate a specific delayed-type hypersensitivity (DTH). The results showed that 6B11 induced a severe foot pad swelling (Mean thickness 0.93mm) as seen in the positive control (P > 0.05) induced by ovarian carcinoma antigen; the 1H12 induced swelling was significantly lower than the positive control (P < 0.05). Both 6B11 and 1H12 expressed the effective responses only to SKOV3, but not to the unrelated target cells. Pathological examinations found that there was marked infiltration of inflammatory cells in foot pad of mice primed by 6B11, while by 1H12, only induced a narrow pathological changes. These observations indicated that there would be some essential differences between 6B11 and 1H12 in the behavior of inducing the specific DTH to SKOK3. And as an idiotypic vaccine, 6B11 would be more powerful against ovarian carcinoma.

Animals↗

[Preparation of monoclonal antibodies against ovarian carcinoma bearing chemical drugs entrapped in liposomes and the cytotoxic tests to carcinoma cells].

Monoclonal antibodies COC166-9 were prepared bearing adriamycin and cis-Platinum entrapped in liposomes respectively as chemoimmunoliposomes MLA and MLP. In vitro growth inhibition of SKOV3 ovarian carcinoma cell lines were tested by MLA and MLP. The results showed that MLA had the same cytotoxic effects as single adriamycin (ADM) in high concentration. However MLA was much stronger than ADM if in low concentration. There was no difference between MLP and single cis-Platinum, of which neither were sensitive to SKOV3 cells.

Antibodies, Monoclonal↗