Biomedical subjects

B Feng

Publications and source records attributed to B Feng.

At least 19 recordsLinked to original sources

PubMed2003

Characterization of surface oxide films on titanium and adhesion of osteoblast.

The relationship between surface characteristics of titanium and initial interactions of titanium-osteoblasts was investigated. Titanium plates were heat-treated in different oxidation atmospheres. The third passage rabbit osteoblasts were cultured on the titanium plates for 24h. After the heat-treatment, the crystal structure of the surface oxide films on titanium was identified using X-ray diffractometer and X-ray photoelectron spectroscopy (XPS). The surface roughness of titanium was measured with a profilometer. The surface energy was obtained by measurement of contact angles and calculation with Owens-Wendt-Kaeble's equation. The amount of surface hydroxyl (OH)(s) groups was examined using XPS. The change of binding energy of the some elements on the substrate surface suggested that the interactions between the cells and the titanium involved chemical reactions. The greater surface roughness, higher surface energy and more surface hydroxyl groups resulted in greater numbers of adhered osteoblasts and higher cell activity. Compared to the acidic hydroxyl (OH)(a) groups in (OH)(s) groups and the dispersion component of the total surface energy, the basic hydroxyl (OH)(b) groups and the polar component play more important roles in the osteoblast-titanium interaction.

Alkaline Phosphatase↗

PubMed2002

Effect of water vapor treatment on apatite formation on precalcified titanium and bond strength of coatings to substrates.

In previous investigations, a simple method, precalcification, was developed for bioactivating titanium. After a titanium sample was precalcified in a boiling saturated Ca(OH)(2) solution and then immersed in a calcium phosphate supersaturated solution, an apatite coating rapidly precipitated onto its surface. In the present study, heat-treatment in water vapor was carried out prior to precalcification. Heat-treatment in water vapor stimulated the chemical reaction between titanium, calcium, and phosphate. Coating properties were improved, and the bond strength of the coating to substrate was enhanced.

Apatites↗

PubMed2002

Carbonate apatite coating on titanium induced rapidly by precalcification.

Chemical treatments have been thought to be promised methods for improving bioactivity of titanium. In this work, the effect of precalcification with boiling saturated Ca(OH)2 solution on bioactivation of titanium was investigated. After precalcification and soaking in supersaturated Ca-P solution (SCP), calcium phosphate rapidly precipitated onto the surfaces of titanium, and after only three days an uniform apatite layer was found up to thickness of a few micrometers. The observation using scanning electron microscopy (SEM) showed that the coating was composed of a number of small crystal grains. The investigation by X-ray energy dispersion spectroscopy (EDS), Fourier transform infrared spectroscopy (FTIR) and X-ray diffraction (XRD) indicated that the coating was Ca-deficient carbonate apatite. Based on the analyses for the surfaces and SCP, a mechanism of precipitation of apatite was proposed in thermal dynamics and kinetics.

Apatites↗

PubMed2002

[The hemolytic site of the basic phospholipase A(2) from Agkistrodon halys pallas].

The gene of the basic phospholipase A(2) from Agkistrodon halys Pallas (BPLA(2) )was mutated site-directedly by polymerase chain reaction (PCR) and the residue Arg(34) of the encloding protein was substituted by Glu and Gln respectively. The mutant gene has been cloned into the expression vector pBLMVL2 and has been expressed in E.coli RR1 effectively. The protein was produced as insoluble inclusion bodies. After partial purification, the inclusion bodies were denatured and renatured into active form, and the renatured recombinant protein was purified by gel-filtration. The expression product has the same enzymatic activity as the denatured-refolded BPLA(2) and its hemolytic activity dropped distinctly, which suggest that the basic residue Arg(34) of BPLA(2) is a crucial amino acid residue during the process of hemolysis.

Agkistrodon↗

PubMed2001

An integrated ten-pump, eight-channel parallel LC/MS system for automated high-throughput analysis of proteins.

An integrated 10-pump eight-channel LC/MS system has been developed for automated high-throughput analysis of intact proteins in recombinant protein purification processes. The key features of the system include (1) a compact 10-pump HPLC module that uses two pumps to generate a binary gradient and 8 pumps to deliver the mixed gradient to eight independent flow channels; (2) a TOF mass spectrometer with an eight-channel multiplexed ESI interface, which records separate data for all eight channels over each HPLC run cycle; and (3) highly automated data processing software that allows unattended calculation of protein molecular weight (in Da) from original mass spectral data (in m/z). This system was used in the routine screening of fractions from preparative scale chromatography to monitor the purification process with the required mass accuracy and throughput. As an example, the production and purification of an acylated protein with a molecular weight of 9 kDa is described. Using this off-line approach, it is practical to fully characterize protein-containing fractions from column chromatography with an overall analytical throughput of 1 min/protein sample with minimum operator involvement.

Autoanalysis↗

PubMed2001

Ca2+ binding site 2 in calcineurin-B modulates calmodulin-dependent calcineurin phosphatase activity.

Calcineurin is the Ca(2+)- and calmodulin-dependent Ser/Thr phosphatase. Human calcineurin-Aalpha and wild-type or mutated calcineurin-Bs were coexpressed in Escherichia coli and purified by calmodulin-Sepharose affinity chromatography. Four calcineurin-B mutants were studied. Each had a single conserved Glu in the 12th position of one EF-hand Ca(2+) binding site replaced by a Lys, resulting in the loss of Ca(2+) binding to that site. Phosphatase activities of the enzymes toward a (32)P-labeled phosphopeptide substrate were measured. Inactivating Ca(2+) binding sites 1, 2, or 3 in calcineurin-B reduced Ca(2+)-dependent phosphatase activity of the enzymes in the absence of calmodulin with the site 2 mutation being most effective. Inactivating Ca(2+) binding site 4 did not change enzyme activity or sensitivity to Ca(2+) in either the absence or presence of calmodulin. The calmodulin-dependent phosphatase activity of the enzymes containing site 1, 2, or 3 mutations in calcineurin-B was also decreased compared to enzyme with wild-type calcineurin-B. Of these enzymes, the one with the site 2 mutation was most profoundly affected as determined by the magnitude of the shift in Ca(2+) concentration dependence. Binding of a fluorescein-labeled calmodulin to the wild-type and the site 2 mutant enzymes was examined using fluorescence polarization measurements. The decrease in Ca(2+) sensitivity for the enzyme with calcineurin-B site 2 inactivated is apparently due to a decrease in the affinity of that enzyme for calmodulin at low Ca(2+) concentrations. These data support a role for Ca(2+) binding site 3 in the carboxyl half of calcineurin-B in transmitting the Ca(2+) signal to calcineurin-A and indicate that site 2 in the amino half of calcineurin-B is critical for enzyme activation.

Affinity Labels↗

PubMed2001

A novel strategy for organ allografts using sublethal (7 Gy) irradiation followed by injection of donor bone marrow cells via portal vein.

A new strategy for organ allografts that does not require recourse to immunosuppressants is established in mice. The strategy includes sublethal (7 Gy) irradiation followed by the injection of donor bone marrow cells (BMCs) via the portal vein (P.V.) and organ allografts 1 day after irradiation. Irradiation doses (< or =7 Gy) are found to allow the recipients to survive without the need to reconstitute the BMCs, as the recipient hematolymphoid cells can gradually recover. One hundred percent of recipients irradiated with 7 Gy followed by either P.V. or i.v. injection of donor BMCs accept organ allografts (the skin, pancreas, and adrenal glands) for more than 1 year. However, organ allograft survival rates decrease when irradiation doses are reduced; the skin graft survival rate of mice treated with 6.5 Gy and P.V. injection of BMCs is 79%, whereas that of mice treated with 6.5 Gy and i.v. injection is 50%, indicating that the P.V. injection of BMCs induces persistent tolerance more effectively than the i.v. injection. H-2 typing reveals that almost all the hematolymphoid cells (>98%) in the peripheral blood and hematolymphoid organs are donor-derived even 1 year after the treatment (7 Gy and P.V.). The T cells are tolerant to both donor-type and host-type MHC determinants. The major mechanism underlying the persistent tolerance induced by this strategy seems to be because of clonal deletion. This simple and safe strategy would be of great advantage for human organ transplantation.

Adrenal Glands↗

PubMed2001

Arginine 454 and lysine 370 are essential for the anion specificity of the organic anion transporter, rOAT3.

Organic anion transporters (OATs) and organic cation transporters (OCTs) mediate the flux of xenobiotics across the plasma membranes of epithelia. Substrates of OATs generally carry negative charge(s) whereas substrates of OCTs are cations. The goal of this study was to determine the domains and amino acid residues essential for recognition and transport of organic anions by the rat organic anion transporter, rOAT3. An rOAT3/rOCT1 chimera containing transmembrane domains 1-5 of rOAT3 and 6-12 of rOCT1 retained the specificity of rOCT1, suggesting that residues involved in substrate recognition reside within the carboxyl-terminal half of these transporters. Mutagenesis of a conserved basic amino acid residue, arginine 454 to aspartic acid (R454D), revealed that this amino acid is required for organic anion transport. The uptakes of p-aminohippurate (PAH), estrone sulfate, and ochratoxin A were approximately 10-, approximately 48-, and approximately 32-fold enhanced in oocytes expressing rOAT3 and were only approximately 2-, approximately 6-, and approximately 5-fold enhanced for R454D. Similarly, mutagenesis of the conserved lysine 370 to alanine (K370A) suggested that K370 is important for organic anion transport. Interestingly, the charge specificity of the double mutant, R454DK370A, was reversed in comparison to rOAT3-R454DK370A preferentially transported the organic cation, MPP(+), in comparison to PAH (MPP(+) uptake/PAH uptake = 3.21 for the double mutant vs 0.037 for rOAT3). These data indicate that arginine 454 and lysine 370 are essential for the anion specificity of rOAT3. The studies provide the first insights into the molecular determinants that are critical for recognition and translocation of organic anions by a member of the organic anion transporter family.

1-Methyl-4-phenylpyridinium↗

PubMed2001

Fast characterization of intact proteins using a high-throughput eight-channel parallel liquid chromatography/mass spectrometry system.

The preparation of protein substrates requires that a large number of chromatographic fractions be analyzed for the presence of reactants, products and by-products. Analyses using linear matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) or single column liquid chromatography/mass spectrometry (LC/MS) have been inadequate because of mass resolution or throughput. Therefore, a high-throughput method employing an eight-channel parallel reverse-phase LC/MS system was developed. This system is capable of screening fractions from preparative ion-exchange chromatography with the required mass accuracy and throughput so that the protein purification process can be monitored in a relatively short period of time. As an example, the purification and analysis of an acylated protein with a molecular weight of 8.9 kDa is described and the detection of a contaminating by-product that differs in size by less than 20 Da is demonstrated. Using the current instrumentation and approach, it is practical to analyze 50 protein-containing fractions from column chromatography in less than 1 hour using parallel LC/MS.

Acyl Carrier Protein↗

PubMed2001

Molecular cloning and characterization of WdPKS1, a gene involved in dihydroxynaphthalene melanin biosynthesis and virulence in Wangiella (Exophiala) dermatitidis.

1,8-Dihydroxynaphthalene (1,8-DHN) is a fungal polyketide that contributes to virulence when polymerized to 1,8-DHN melanin in the cell walls of Wangiella dermatitidis, an agent of phaeohyphomycosis in humans. To begin a genetic analysis of the initial synthetic steps leading to 1,8-DHN melanin biosynthesis, a 772-bp PCR product was amplified from genomic DNA using primers based on conserved regions of fungal polyketide synthases (Pks) known to produce the first cyclized 1,8-DHN-melanin pathway intermediate, 1,3,6,8-tetrahydroxynaphthalene. The cloned PCR product was then used as a targeting sequence to disrupt the putative polyketide synthase gene, WdPKS1, in W. dermatitidis. The resulting wdpks1Delta disruptants showed no morphological defects other than an albino phenotype and grew at the same rate as their black wild-type parent. Using a marker rescue approach, the intact WdPKS1 gene was then successfully recovered from two plasmids. The WdPKS1 gene was also isolated independently by complementation of the mel3 mutation in an albino mutant of W. dermatitidis using a cosmid library. Sequence analysis substantiated that WdPKS1 encoded a putative polyketide synthase (WdPks1p) in a single open reading frame consisting of three exons separated by two short introns. This conclusion was supported by the identification of highly conserved Pks domains for a beta-ketoacyl synthase, an acetyl-malonyl transferase, two acyl carrier proteins, and a thioesterase in the deduced amino acid sequence. Studies using a neutrophil killing assay and a mouse acute-infection model confirmed that all wdpks1Delta strains were less resistant to killing and less virulent, respectively, than their wild-type parent. Reconstitution of 1,8-DHN melanin biosynthesis in a wdpks1Delta strain reestablished its resistance to killing by neutrophils and its ability to cause fatal mouse infections.

Amino Acid Sequence↗

PubMed2001

Successful allogeneic bone marrow transplantation (BMT) by injection of bone marrow cells via portal vein: stromal cells as BMT-facilitating cells.

We examined the importance of the coadministration of bone marrow (BM) stromal cells with BM cells via the portal vein. A significant increase in the number of day-14 colony-forming unit-spleen (CFU-S) was observed in the recipient mice injected with hemopoietic stem cells (HSCs) along with donor BM stromal cells obtained after three to four weeks of culture. Histological examination revealed that hematopoietic colonies composed of both donor hemopoietic cells and stromal cells coexist in the liver of these mice. However, when donor HSCs plus BM stromal cells were administered i.v., neither the stimulatory effects on CFU-S formation nor the hemopoietic colonies in the recipient liver were observed. These findings suggest that the interaction of HSCs with stromal cells in the liver is the first crucial step for successful engraftment of allogeneic HSCs. It is likely that donor stromal cells and HSCs trapped in the liver migrate into the recipient BM and spleen, where they form CFU-BM and CFU-S, respectively.

Animals↗

PubMed2001

Functional characteristics and steroid hormone-mediated regulation of an organic cation transporter in Madin-Darby canine kidney cells.

Organic cation transporters (OCT1-3) play an important role in renal elimination of many drugs. The goals of this study were to 1) identify a cell culture model which constitutively expressed OCT2 that could be used to study the characteristics and regulation of this transporter, and 2) to study the mechanisms by which xenobiotics and hormones regulate the activity of OCT2. We characterized the endogenous organic cation transporter (OCT) activity in Madin-Darby canine kidney (MDCK) cells. The activity was localized to the basolateral membrane and was pH and membrane potential-dependent. The uptake of the model organic cation, tetraethylammonium, was saturable (Km, 19.5 +/- 4.6 microM; Vmax, 350 +/- 19.4 pmol/mg of protein/10 min) and was inhibited by known OCT inhibitors (e.g., cimetidine and quinidine). A cDNA fragment (711 base pairs) isolated by reverse transcriptase-polymerase chain reaction (RT-PCR) was greater than 83% identical to OCT2 cDNAs from mammalian species; no OCT1 or OCT3 was detected by RT-PCR, suggesting that OCT2 may be the primary basolateral OCT in MDCK. OCT2 mRNA levels were increased significantly following exposure of MDCK to the steroid hormones, dexamethasone (2.0-fold), hydrocortisone (2.4-fold), and testosterone (1.8-fold) with comparable increases in activity. Other compounds tested, including the cytochrome P450 inducers, rifampicin, phenobarbital, and phenytoin, and the OCT substrates, verapamil and metformin, had no inducing effects. Collectively, these data indicate that MDCK can serve as a useful and convenient tool in screening candidate drugs for interaction with OCT2 and for studying the regulation of this transporter. Furthermore, our data demonstrate that steroid hormones induce the transcription of OCT2 in the kidney.

Animals↗

PubMed2001

Murine antibody against E2 can capture hepatitis C virus in vitro.

OBJECTIVE: To find neutralizing antibody candidates against hepatitis C virus (HCV) infection. METHODS: We constructed two eukaryotic expression vectors which contained the E1 and E2 gene of HCV, and detected their expression in mammalian cells with transient expression. BALB/c mice were given subculaneous injections of constructed vectors combined with the IL-2 gene intraepidermally and evaluated for induced humoral immune responses by enzyme-linked immunosorbent assay (ELISA). We used an antibody-virus interaction assay to analyze the interaction of the antisera and HCV viral particles in vitro. RESULTS: Anti E1 and anti-E2 antisera were obtained from immunized mice. The serum of mice immunized with the E2 gene immunoprecipitated the HCV isolate in source serum and reacted with the isolates unrelated to the original one. CONCLUSIONS: Anti-E2 antibody in induced mice can cross-reactively capture HCV particles, highlighting the possibility of generating broadly reactive anti-E2 antibodies.

Animals↗

PubMed2001

[Study on the transformation from myelodysplastic syndromes into acute leukemias].

OBJECTIVE: To study the patterns of transformation from myelodysplastic syndromes (MDS) into acute leukemias (AL). METHODS: Leukemic transformation of MDS patients was dynamically followed up and the clinical manifestations, peripheral blood and bone marrow pictures, karyotypes, immunophenotypes, response to treatment and prognosis of post MDS acute leukemia (postMDS-AL) were observed. RESULTS: During the past eight year and seven months, 21 (13.91%) of 151 MDS patients progressed to overt leukemia with a median interval of 5 (1 - 21) months. There were no significant differences among the rates of leukemia from RA, RAEB and RAEB-t groups. The transformation was developed either gradually or rapidly. There were five parameters related to the leukemic transformation: under 40 years of age, pancytopenia, more than 0.15 blasts in bone marrow, at least two types of abnormal karyotype and combined chemotherapy. All of the 21 post MDS-AL were acute myeloid leukemia (AML); and most of them were M(2), M(4) and M(5). Two (9.52%) post MDS-AML developed extramedullary infiltration. Leukopenia was found in 47.62% of patients. Two third of the patients, whose bone marrows were generally hypercellular, showed neutropenias. After evolving into AML, 8 (47.06%) patients developed abnormal karyotypes. High expression of immature myeloid antigens, including CD(33) (49.83 +/- 24.50)%, CD(13) (36.38 +/- 33.84)%, monocytic antigen CD(14) (38.50 +/- 24.60)%, and stem cell marker CD(34) (34.67 +/- 30.59)% were found on bone marrow mononuclear cells of post MDS-AML cases. In some cases, lymphoid antigens, such as CD(5), CD(7), CD(9) and CD(19) were coexisted with myeloid antigens. A low complete remission rate (31.25%) and short survival duration with median survival of 6 (1 - 28) months were found in patients with post MDS-AML treated by induction therapy. CONCLUSION: MDS was at high risk of evolving into AML, either gradually or rapidly. Patients with post MDS-AML had specific biologic features and worse prognoses.

Acute Disease↗

PubMed2001

[Study on the working noise in BYPC and the effects caused by working noise on the workers' vestibular and auditory function].

OBJECTIVE: In order to observe the kinds and intensity of the working noise of Yansan Petrochemical Co. and the effects caused by the working noise on the workers' vestibular and auditory function. METHOD: The intensity and frequency of the working noise were recorded by exactolnoisemeter in the workshop. One hundred and seventeen workers were tested in routine pure-tone tested method. The SPVN and ABR were tested within fifty-one workers of all. RESULT: The working noise of Yansan Petrochemical Co. belongs to the broad band and steady noise. The intensity of the working noise were during 85.7-104.0 dB(A) and the main frequency were during 1-8 kHz. About 59 percent workers who exposed to the working noise had hearing loss. The most hearing-loss were in the high frequency. The hearing-loss of speech frequency were slight. Workers who have more six years standing have obviously increased hearing-loss than the workers who have less five years standing. There were significant differences the ABR thresholds and wave-interval between the tested and controlled groups. The SPVN and CP were abnormal in more than 17.4 percent workers with hearing loss. CONCLUSION: The working noise of Yansan Petrochemical Co. belongs to the broad band and steady noise. Working noise can lead to workers' hearing loss of certain degrees who exposed in the noise for a long time. Obvious correlation was not defined between the hearing-loss and the abnormal vestibular response group.

Adult↗

PubMed2001

The preliminary clinical observation of array multifocal intraocular lens implantation.

PURPOSE: To evaluate the clinical effects of implantation of Array multifocal intraocular lenses. METHODS: Thirty-one cases (37 eyes) of cataract patients, including 15 males (19 eyes) and 16 females (18 eyes), were involved in this study. All patients underwent standard phacoemulsification with Array multifocal intraocular lens implantation. The complications during operation, postoperative distant visual acuity, near visual acuity, corneal curvature and visual symptoms were observed. RESULTS: The mean value of best postoperative visual acuity was recorded as follows: uncorrected distant visual acuity was 0.8, the best-corrected distant visual acuity was 0.9, uncorrected near visual acuity was 0.5, near visual acuity with distant-corrected was 0.6, the best-corrected near visual acuity was 0.9. The astigmatism of cornea was less than 1.5 D pre-operatively and post-operatively. One patient complained of glare. CONCLUSION: Array multifocal intraocular lens can provide good distant and near visual acuity. With observation of more cases and follow-up of longer time, we can draw a further conclusion.

Aged↗

PubMed2001

Comparison of trans-scleral fixation of PMMA and foldable intraocular lens in children.

PURPOSE: To observe the difference of the effects of PMMA and foldable intraocular lenses (IOLs) trans-sclerally fixed in pediatric eyes. METHODS: Thirty-two children (43 eyes) who had undergone trans-scleral fixation of IOL were retrospected, of whom 5 children were implanted PMMA IOL in both eyes, 6 children were implanted PMMA IOL in one eye and foldable IOL in the other eye, 12 children were implanted foldable IOL in one eye and 9 children were implanted PMMA IOL in one eye. Mean age was 5.3 years (range 2.5-12 years). Twelve children had traumatic cataract and the others congenital cataract before lens extraction. RESULTS: Foldable group (18 eyes): Mean follow-up was 12.1 months. Visual acuity (VA): compared with the best corrected VA before IOL fixation, postoperative best corrected VA improved in 16 eyes, remained unchanged in 2 eyes. In 14 eyes, one or two stitches were needed to seal the incision. COMPLICATIONS: Severe anterior chamber reaction was seen in 3 eyes. Intraocular bleeding was found in 3 eyes. IOL decentration was detected in 1 eye. Iris capture of IOL was seen in one eye. PMMA group (25 eyes): Mean follow-up was 20.3 months. Visual acuity (VA): compared with the best corrected VA before IOL fixation, postoperative best corrected VA improved in 19 eyes, remained unchanged in 5 eyes and got worse in one eye. In 24 eyes, one to three stitches were needed to seal the incision. COMPLICATIONS: Severe anterior chamber reaction was seen in 5 eyes. Intraocular bleeding was found in 4 eyes. IOL decentration was seen in one eye. Iris capture of IOL was seen in 3 eyes. Intraocular pressure elevated in one eye. CONCLUSION: Our study shows that trans-scleral fixation of IOL is a safe procedure in pediatric eyes. Foldable IOL showed similar effect compared with PMMA IOL in pediatric trans-scleral fixation.

Anterior Chamber↗

PubMed2001

[Effects of acute infrasound exposure on vestibular and auditory functions and the ultrastructural changes of inner ear in the guinea pig].

OBJECTIVE: To define the effects of acute infrasound exposure on vestibular and auditory functions and the ultrastructural changes of inner ear in guinea pigs. METHODS: The animals involved in the study were exposed to 8 Hz infrasound at 135dB SPL for 90 minutes in a reverberant chamber. The sinusoidal pendular test (SPT), auditory brainstem response (ABR) and distortion product otoacoustic emissions (DPOAE) were respectively detected pre-exposure and at 0(within 2 hrs), 2 and 5 day after exposure. The ultrastructures of the inner ear were observed by scanning electron microscopy. RESULTS: The slow-phase velocity and the frequency of the vestibular nystagmus elicited by sinusoidal pendular test (SPT) declined slightly following infrasound exposure, but the changes were not significant (P > 0.05). No differences in the ABR thresholds, the latencies and the interval peak latencies of I, III, V waves were found between the normal and the experimental groups, and among experimental groups. The amplitudes of DPOAE at any frequency declined remarkably in all experimental groups. The ultrastructures of the inner ear were damaged to different extent. CONCLUSION: Infrasound could transiently depress the excitability of the vestibular end-organs, decrease the function of OHC in the organ of Corti and cause damage to the inner ear of guinea pigs.

Animals↗