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X D Li

Publications and source records attributed to X D Li.

At least 37 records · Page 2Linked to original sources
PubMed2001

High-yield expression and functional analysis of Escherichia coli glycerol-3-phosphate transporter.

The glycerol-3-phosphate (G3P) transporter, GlpT, from Escherichia coli mediates G3P and inorganic phosphate exchange across the bacterial inner membrane. It possesses 12 transmembrane alpha-helices and is a member of the Major Facilitator Superfamily. Here we report overexpression, purification, and characterization of GlpT. Extensive optimization applied to the DNA construct and cell culture has led to a protocol yielding approximately 1.8 mg of the transporter protein per liter of E. coli culture. After purification, this protein binds substrates in detergent solution, as measured by tryptophan fluorescence quenching, and its dissociation constants for G3P, glycerol-2-phosphate, and inorganic phosphate at neutral pH are 3.64, 0.34, and 9.18 microM, respectively. It also shows transport activity upon reconstitution into proteoliposomes. The phosphate efflux rate of the transporter in the presence of G3P is measured to be 29 micromol min(-1) mg(-1) at pH 7.0 and 37 degrees C, corresponding to 24 mol of phosphate s(-1) (mol of protein)(-1). In addition, the glycerol-3-phosphate transporter is monomeric and stable over a wide pH range and in the presence of a variety of detergents. This preparation of GlpT provides ideal material for biochemical, biophysical, and structural studies of the glycerol-3-phosphate transporter.

Biological Transport↗
PubMed2001

Heavy metal speciation and leaching behaviors in cement based solidified/stabilized waste materials.

A circuit board printing factory sludge containing high concentrations of copper, zinc and lead was stabilized and solidified (S/S) with different portions of ordinary Portland cement (OPC) and pulverized fly ash (PFA). The chemical speciation and leaching behavior of heavy metals in these cement-based waste materials were studied by different sequential extraction procedures, standard toxicity characteristic leaching procedure (TCLP) and progressive TCLP tests. The sequential extraction results showed that more than 80% of Cu, Pb and Zn were associated with Fraction 2 (weak acid soluble, extracted with 1M NaOAc at pH 5.0 with a solid to liquid ratio of 1:60). This indicated that the heavy metals could exist in the S/S matrix as metal hydrated phases or metal hydroxides precipitating on the surface of calcium silicate hydrates (C-S-H), PFA and sludge particles. The progressive TCLP test results and MINTEQA2 calculation also showed the importance of Cu and Zn oxides during the leaching process. The leaching behaviors of these metals in the S/S waste materials were mainly controlled by the alkaline nature and acid buffering capacity of the S/S matrix. During the progressive TCLP tests, the alkaline conditions and acid buffering capacity of the matrix decreased with the dissolution of calcium hydroxide and C-S-H, therefore, the leaching of heavy metals in the S/S waste materials increased. The leaching of heavy metals in the S/S materials can be considered as a pH dependent and corresponding metal hydroxide solubility controlled process.

Hydrogen-Ion Concentration↗
PubMed2001

Monomeric state and ligand binding of recombinant GABA transporter from Escherichia coli.

The gamma-aminobutyric acid (GABA) transporter from Escherichia coli was homologously overexpressed and purified to homogeneity with a yield of 1.0 mg per liter culture. The purification procedure consists of a cobalt affinity column, proteolytic cleavage of His- and myc-tags, and size-exclusion chromatography. The purified transporter exists as a monomer in FOS-Choline 12 detergent, with a Stokes radius of 45 A for the protein-detergent complex. In detergent solution the protein binds substrates, as indicated by tryptophan fluorescence quenching. Its dissociation constants (K(d)) for GABA, muscimol and nipecotic acid are 13.8, 13.3 and 27.9 microM, respectively. This protein preparation provides ideal starting materials for future biochemical, biophysical and structural studies of the GABA transporter.

Bacterial Proteins↗
PubMed2001

Homeostasis of naive and memory T cell subpopulations in peripheral blood and lymphoid tissues in the context of human immunodeficiency virus infection.

To understand the nature of naive and memory T cell depletion in human immunodeficiency virus (HIV) immunopathogenesis, their homeostasis in peripheral blood (PB) and lymph node (LN) compartments of HIV-infected patients was examined. Although the percentage of naive CD4+ cells was higher in LN than in PB mononuclear cells (LNMC and PBMC, respectively), the memory cells were higher in PBMC than in LNMC. The ratio of naive:memory CD4+ cells from PB positively correlated with that in LNs and with the absolute CD4+ cell counts and recall antigen responses, and the ratio inversely correlated with the cellular virus load from the corresponding compartment. These findings indicate that although the pattern of naive and memory cells in the LN and PB compartments appear divergent, their relationship is nonrandom and is significant. The naive&rcolon;memory ratio in PB appears to reflect the lymphoid microenvironment and may potentially be useful as a surrogate marker for treatment efficacy and immune reconstitution.

Adult↗
PubMed2001

Chemokine/CD4 receptor density ratios correlate with HIV replication in lymph node and peripheral blood of HIV-infected individuals.

OBJECTIVES: Lymphoid tissue is a major reservoir for virus replication in HIV-infected subjects. The relationship of CCR5 and CXCR4 coreceptor density and HIV replication in peripheral blood mononuclear cells (PBMC) and lymph node (LN) mononuclear cells (LNMC) of HIV-infected subjects was examined. METHODS: PBMC and cervical LNMC from 12 HIV-infected patients were examined for virological and immunological parameters including chemokine receptor density, HIV plasma and cellular viral load, coreceptor usage and CD38/HLA-DR expression. RESULTS: The number of CCR5 and CXCR4 molecules on CD4 lymphocytes in the LN were significantly higher than in PBMC. In contrast the number of CD4 molecules/CD4 T cell was higher in PBMC than in LNMC. The CXCR4/CD4 and CCR5/CD4 ratios in the LN were significantly higher than in the PBMC. This was associated with a cellular viral load in the LN that was approximately 110-fold higher than in PBMC. The absolute number of coreceptor molecules per cell did not correlate with the viral load. However, the CCR5/CD4 and CXCR4/CD4 ratios in the LN positively correlated with HIV cellular and plasma RNA. Characterization of the viral isolates suggested an association between clinical isolates using a distinct coreceptor and the upregulation of the corresponding chemokine receptor. CONCLUSIONS: The ratios of chemokine receptors to CD4 molecules in CD4 T cells from LN is higher than in PBMC and may account for the relative difference in cellular viral load in these compartments. Additionally, the coreceptor/CD4 ratios, particularly in the lymphoid tissue, were highly related to HIV replication.

Adult↗
PubMed2001

Profiling of differential gene expression in activated, allergen-specific human Th2 cells.

Th2 cells play a pivotal role in the pathogenesis of allergic diseases, including asthma, but the molecular basis of the Th1/Th2 dichotomy and the precise molecular pathways leading to Th2-dominant immune responses are still unclear. To this end, we have combined suppression subtractive hybridization (SSH) and high throughput analysis of cDNA arrays spotted with IMAGE clones to determine the profile of differential gene expression in human allergen-specific Th2 cells. Allergen-stimulated Th2 cells were used as the tester, and either resting Th2 cells or stimulated Th1 cells were used as the driver. SSH was used to equalize different mRNA levels and remove common sequences between the tester and the driver. Comparison of cDNA arrays probed with subtracted tester and non-subtracted driver provided a profile of Th2-selective gene expression. Analysis of 77 sequence-confirmed and differentially expressed genes in Th2 cells showed predominant EST sequences, representing 80% of sequences analyzed. The pattern of gene expression in 19 selected sequences was further analyzed in additional Th1 and Th2 clones. A total of 15 sequences showed predominant expression in Th2 cells, while the remaining four EST sequences showed no detectable amplification signal. The database containing Th2-selective genes will further our understanding of Th2 cell function and the genetic basis of allergic diseases.

Allergens↗
PubMed2001

The treatment of osteomyelitis with gentamicin-reconstituted bone xenograft-composite.

We have developed a new drug-delivery system using reconstituted bone xenograft to treat chronic osteomyelitis. This material, which has the capabilities of osteoinduction and osteoconduction, was supplemented with up to 2000 times the minimum inhibitory concentration of gentamicin against Staphylococcus aureus to prepare a gentamicin-reconstituted bone xenograft-composite (G-RBX-C). In a rabbit model, we evaluated the release of gentamicin from this composite in vivo, its capability for induction of ectopic bone and the repair of segmental defects of the radius. There was a high level of concentration of antibiotics, which was sustained for at least ten days. In the study of induction of ectopic bone, there was abundant woven bone in the G-RBX-C group two weeks after operation. At 16 weeks after implantation of G-RBX-C the radial defects had been repaired, with the formation of lamellar bone and recanalisation of the marrow cavity. Our findings suggest that G-RBX-C may be useful in the treatment of chronic osteomyelitis.

Animals↗
PubMed2001

[Construction of human bone morphogenetic protein-2 expressing eukaryotic vector].

OBJECTIVE: To construct human bone morphogenetic protein-2(hBMP-2) expressing eukaryotic vector and observe whether it can be expressed in eukaryotic cells. METHODS: pUC19 was digested with Sal I and Xba I. The resulting Sal I-Xba I fragment (1.24 kb) which contains the full length of human BMP-2 cDNA was separated on agarose gel and ligated into eukaryotic expression vector pcDNA3 digested with XhoI and XbaI. The recombinant pcDNA3-hBMP-2 plasmid was transferred into fibroblasts cell line NIH3T3. The stable expression of hBMP-2 in the positive cells G418 selected was determined by in situ hybridization and immunohistochemical analysis. RESULTS: The two fragments digested from recombinant pcDNA3-hBMP-2 plasmid by EcoR I and Xba I represented 1.3 kb and 5.38 kb respectively by agarose electrophoresis, meanwhile the Xho I site was disappeared in pcDNA3-hBMP-2 indicating the successful construction of recombinant pcDNA3-hBMP-2 plasmid. Stable expression of hBMP-2 in pcDNA3-hBMP-2 transfected cells was confirmed by in situ hybridization and immunohistochemical analysis. CONCLUSION: hBMP-2 expressing eukaryotic vector is successfully constructed and can be expressed in eukaryonic cells.

3T3 Cells↗
PubMed2001

Inhibitory role of GDP on saikosaponin (I) stimulated enzymes secretion and rising of [Ca2+]i in rat pancreatic acini.

AIM: To study the role of guanosine diphosphate (GDP) on enzyme secretion and rising of [Ca2+]i in saikosaponin (I) [SA(I)] stimulated rat pancreatic acini. METHODS: Cell membrane of isolated rat pancreatic acini were permeabilized using streptolysin O (SLO). Enzymes secretions were indicated by detecting total protein secretions. Intracellular Ca2+ ([Ca2+]i) was measured using Fluo-3 in SPEX spectrofluorimeter. RESULTS: The inhibition of GDP on SA(I) stimulated enzymes secretion increased with increasing GDP concentration. There were two peaks in the time course of increase in [Ca2+]i evoked by SA(I) 10 micromol/L. After adding GDP 5 mmol/L, [Ca2+]i rose gradually without the two peaks. In permeabilized acini, the accumulation of enzymes stimulated by SA(I) in 30 min reduced by 57 % compared with intact acini. GDP 5 mmol/L decreased the initial rate of secretion. CONCLUSION: Inhibition of GDP on increase in [Ca2+]i reduces SA(I) stimulated enzymes secretion in pancreatic acini.

Animals↗
PubMed2000

The interaction between the regulatory light chain domains on two heads is critical for regulation of smooth muscle myosin.

Recent findings have suggested that the interaction between the two heads is critical for phosphorylation-dependent regulation of smooth muscle myosin. We hypothesized that the interaction between the two regulatory light chains on two heads of myosin dictates the regulation of myosin motor function. To evaluate this notion, we engineered and characterized smooth muscle heavy meromyosin (HMM), which is composed of one entire HMM heavy chain and one motor domain truncated heavy chain containing the S2 rod and regulatory light chain (RLC) binding site, as well as the bound RLC (SMDHMM). SMDHMM was inactive for both actin-translocating activity and actin-activated ATPase activity in the dephosphorylated state, demonstrating that the interaction between the two RLC domains on the two heads and/or a motor domain and a RLC domain in a distinct head is sufficient for the inhibition of smooth muscle myosin motor activity. When phosphorylated, SMDHMM was activated for both actin-translocating activity and actin-activated ATPase activity; however, these activities were lower than those of double-headed HMM, implying partial release of inhibition by phosphorylation in SMDHMM and/or cooperativity between the two heads of smooth muscle myosin. The present results indicate that the RLC domain is critical for phosphorylation-dependent regulation of smooth muscle myosin motor activity. On the other hand, similar to double-headed HMM, SMDHMM showed both "folded" and "extended" conformations, and the ratio of those conformations is dependent on ionic strength, suggesting that the RLC domain is sufficient to regulate the conformational transition in myosin.

Actins↗
PubMed2000

Toxicity of cinnamomin--a new type II ribosome-inactivating protein to bollworm and mosquito.

The toxicity of cinnamomin, a new type II ribosome-inactivating protein purified from the seeds of camphor tree (Cinnamomum camphora), to bollworm (Helicoverpa armigera) and mosquito (Culex pipines pallens) during larval stage was tested. The LC50 of cinnamomin to bollworm larvae fed on diet containing cinnamomin was 1839 ppm and the LC50 to larvae of mosquito was 168 ppm. The gut extract of bollworm larvae could apparently hydrolyze cinnamomin. The inhibition of protein synthesis by cinnamomin was tested in in vitro translation system of bollworm larvae, and its LC50 was determined to be approx. 14 nM. Bollworm larvae ribosome treated with cinnamomin produced a specific RNA fragment (R-fragment) characterized on urea-denatured polyacrylamide gel. Evidence was provided that hidden breaks exist in the largest ribosomal RNA of bollworm larvae.

Algal Proteins↗
PubMed2000

Optical coherence tomography: advanced technology for the endoscopic imaging of Barrett's esophagus.

BACKGROUND AND STUDY AIMS: Endoscopic optical coherence tomography (OCT) is an emerging medical technology capable of generating high-resolution cross-sectional imaging of tissue microstructure in situ and in real time. We assess the use and feasibility of OCT for real-time screening and diagnosis of Barrett's esophagus, and also review state-of-the-art OCT technology for endoscopic imaging. MATERIALS AND METHODS: OCT imaging was performed as an adjunct to endoscopic imaging of the human esophagus. Real-time OCT (13-microm resolution) was used to perform image-guided evaluation of normal esophagus and Barrett's esophagus. Beam delivery was accomplished with a 1-mm diameter OCT catheter-probe that can be introduced into the accessory channel of a standard endoscope. Different catheter-probe imaging designs which performed linear and radial scanning were assessed. Novel ultrahigh-resolution (1.1-microm resolution) and spectroscopic OCT techniques were used to image in vitro specimens of Barrett's esophagus. RESULTS: Endoscopic OCT images revealed distinct layers of normal human esophagus extending from the epithelium to the muscularis propria. In contrast, the presence of gland- and crypt-like morphologies and the absence of layered structures were observed in Barrett's esophagus. All OCT images showed strong correlations with architectural morphology in histological findings. Ultrahigh-resolution OCT techniques achieved 1.1-microm image resolution in in vitro specimens and showed enhanced resolution of architectural features. Spectroscopic OCT identified localized regions of wavelength-dependent optical scattering, enhancing the differentiation of Barrett's esophagus. CONCLUSIONS: OCT technology with compact fiberoptic imaging probes can be used as an adjunct to endoscopy for real-time image-guided evaluation of Barrett's esophagus. Linear and radial scan patterns have different advantages and limitations depending upon the application. Ultrahigh-resolution and spectroscopic OCT techniques improve structural tissue recognition and suggest future potential for resolution and contrast enhancements in clinical studies. A new balloon catheter-probe delivery device is proposed for systematic imaging and screening of the esophagus.

Barrett Esophagus↗
PubMed1999

The order of calcium and phosphate ion deposition on chemically treated titanium surfaces soaked in aqueous solution.

The mechanism of apatite deposition on chemically treated Ti surfaces still is being studied. In this study, simulated body fluid, calcium aqueous solution, phosphate aqueous solution, and accelerated calcification solution are used as media to investigate the order of calcium and phosphate ion deposition on chemically treated Ti surfaces. The results of inductively coupled plasma spectra, scanning electron microscopy, and energy dispersive X-ray analysis show that calcium deposition is the prerequisite for phosphate ion deposition.

Biocompatible Materials↗
PubMed1999

Indoor air quality investigation on commercial aircraft.

Sixteen flights had been investigated for indoor air quality (IAQ) on Cathay Pacific aircraft from June 1996 to August 1997. In general, the air quality on Cathay Pacific aircraft was within relevant air quality standards because the average age of aircraft was less than 2 years. Carbon dioxide (CO2) levels on all flights measured were below the Federal Aviation Administration (FAA) standard (30,000 ppm). The CO2 level was substantially higher during boarding and de-boarding than cruise due to low fresh air supply. Humidity on the aircraft was low, especially for long-haul flights. Minimum humidity during cruise was below the American Society of Heating, Refrigerating, and Air-Conditioning Engineers (ASHRAE) minimum humidity standard (20%). The average temperature was within a comfortable temperature range of 23 +/- 2 degrees C. The vertical temperature profile on aircraft was uniform and below the International Standard Organization (ISO) standard. Carbon monoxide levels were below the FAA standard (50 ppm). Trace amount of ozone detected ranged from undetectable to 90 ppb, which was below the FAA standard. Particulate level was low for most non-smoking flights, but peaks were observed during boarding and de-boarding. The average particulate level in smoking flights (138 micrograms/m3) was higher than non-smoking flights (7.6 micrograms/m3). The impact on IAQ by switching from low-mode to high-mode ventilation showed a reduction in CO2 levels, temperature, and relative humidity.

Air Pollution, Indoor↗
PubMed1999

Preliminary study on HA coating percutaneously implanted in bone.

A comparative investigation on the possibility of hydroxyapatite (HA) coating and pure Ti column to form biological sealing with skin tissue was completed in this study. HA coating and pure Ti column were percutaneously implanted in the tibia of rabbits. Compared with titanium (Ti) implant, HA coating forms epithelial sealing with skin tissue at 6 weeks postoperatively, while the Ti implant may loosen from the implanted site and be lost. The Ti column loosing rate at this time was 50%. However, once the Ti implant becomes fixed with the bone tissue, it can form epithelial sealing with skin tissue just like the HA coating, at 8 weeks postoperatively. At 8 weeks postoperatively, the epithelial sealing is not destroyed in spite of the fact that the HA coating is biodegraded. Our results show that the HA coating can become fixed with the bone faster than the Ti, which is beneficial for epithelial sealing formation. The main role of HA coating for epithelial sealing is beneficial for sealing at the initial period after it is implanted.

Animals↗