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Biomedical subjects

Lu Wang

Publications and source records attributed to Lu Wang.

At least 127 records · Page 7Linked to original sources

Adaptation to chronic length change in explanted airway smooth muscle.

It has been shown that airway smooth muscle in vitro is able to maintain active force over a large length range by adaptation in the absence of periodic stimulations at 4 degrees C (Wang L, Paré PD, and Seow CY. J Appl Physiol 90: 734-740, 2001). In this study, we show that such adaptation also takes place at body temperature and that long-term adaptation results in irreversible functional change in the muscle that could lead to airway hyperresponsiveness. Rabbit tracheal muscle explants were passively maintained at shortened and in situ length for 3 and 7-8 days in culture media; the length-tension relationship was then examined. The length associated with maximal force generation decreased by 10.5 +/- 3.8% (SE) after 3 days and 37.7 +/- 8.5% after 7 or 8 days of passive shortening. At day 3, the left shift in the length-tension curve due to adaptation at short lengths was reversible by readapting the muscle at a longer length. The shift was, however, not completely reversible after 7 days. The results suggest that long-term adaptation of airway smooth muscle could lead to increased muscle stiffness and force-generating ability at short lengths. Under in vivo condition, this could translate into resistance to stretch-induced relaxation and excessive airway narrowing.

Adaptation, Physiological↗

Kinetics and mechanisms of S(IV) reductions of bromite and chlorite ions.

The reaction of bromite with aqueous S(IV) is first order in both reactants and is general-acid catalyzed. The reaction half-lives vary from 5 ms (p[H+] 5.9) to 210 s (p[H+] 13.1) for 0.7 mM excess S(IV) at 25 degrees C. The proposed mechanism includes a rapid reaction (k(1) = 3.0 x 10(7) M(-1) s(-1)) between BrO(2)(-) and SO(3)(2-) to form a steady-state intermediate, (O(2)BrSO(3))(3-). General acids assist the removal of an oxide ion from (O(2)BrSO(3))(3-) to form OBrSO(3)(-), which hydrolyzes rapidly to give OBr(-) and SO(4)(2-). Subsequent fast reactions between HOBr/OBr(-) and SO(3)(2-) give Br(-) and SO(4)(2-) as final products. In contrast, the chlorite reactions with S(IV) are 5-6 orders of magnitude slower. These reactions are specific-acid, not general-acid, catalyzed. In the proposed mechanism, ClO(2)(-) and SO(3)H(-)/SO(2) react to form (OClOSO(3)H)(2)(-) and (OClOSO(2))(-) intermediates which decompose to form OCl(-) and SO(4)(2-). Subsequent fast reactions between HOCl/OCl(-) and S(IV) give Cl- and SO(4)(2-) as final products. SO(2) is 6 orders of magnitude more reactive than SO(3)H-, where k(5)(SO(2)/ClO(2)(-)) = 6.26 x 10(6) M(-1) s(-1) and k(6)(SO(3)H(-)/ClO(2)(-)) = 5.5 M(-1) s(-1). Direct reaction between ClO(2)(-) and SO(3)(2-) is not observed. The presence or absence of general-acid catalysis leads to the proposal of different connectivities for the initial reactive intermediates, where a Br-S bond forms with BrO(2)(-) and SO(3)(2-), while an O-S bond forms with ClO(2)(-) and SO(3)H-.

Journal Article↗

Identification of eight genes encoding chemokine-like factor superfamily members 1-8 (CKLFSF1-8) by in silico cloning and experimental validation.

TM4SF11 is only 102 kb from the chemokine gene cluster composed of SCYA22, SCYD1, and SCYA17 on chromosome 16q13. CKLF maps on chromosome 16q22. CKLFs have some characteristics associated with the CCL22/MDC, CX3CL1/fractalkine, CCL17/TARC, and TM4SF proteins. Bioinformatics based on CKLF2 cDNA and protein sequences in combination with experimental validation identified eight novel genes designated chemokine-like factor superfamily members 1-8 (CKLFSF1-8). CKLFSF1-8 form gene clusters; the sequence identities between CKLF2 and CKLFSF1-8 are from 12.5 to 39.7%. Most of the CKLFSFs have alternative RNA splicing forms. CKLFSF1 has a CC motif and higher sequence similarity with chemokines than with any of the other CKLFSFs. CKLFSF8 shares 39.3% amino acid identity with TM4SF11. CKLFSF1 links the CKLFSF family with chemokines, and CKLFSF8 links it with TM4SF. The characteristics of CKLFSF2-7 are intermediate between CKLFSF1 and CKLFSF8. This indicates that CKLFSF represents a novel gene family between the SCY and the TM4SF gene families.

Alternative Splicing↗

Molecular cloning and characterization of four isoforms of mCKLF, mouse homologues of human chemokine-like factor.

Chemokine-like factor1 (CKLF1), and its three isoforms (CKLF2, 3 and 4), are recently identified human cytokines. CKLF1 is a potent chemoattractant for human leukocytes and can stimulate inflammation and the regeneration of murine skeletal muscle. CKLF2 can promote proliferation and differentiation of C2C12 muscle cells directly by inducing expression of myogenin and activating transcription factors. In the present study, we cloned CKLF murine homologues, and based on their biological and structural features, named them murine chemokine-like factor 2, 4, 5 and 6 (mCKLF2, 4, 5 and 6). mCKLF2, 4, 5 and 6 encode 152, 120, 122 and 86 amino-acid proteins, respectively. mCKLFs map to mouse chromosome 8 and have high sequence similarity to human CKLFs. Compared to human CKLFs, which have a CC motif in the C-terminal region, mCKLF2 and 4 contain a CX3C motif. Using a PCR-based approach, it appeared mCKLF2 and 5 mRNA were highly expressed in adult testis, while mCKLF4 mRNA was detected only in differentiated C2C12 cells, a pattern different from human CKLFs. Conditioned media from COS-7 cells transfected with mCKLF2 and 4 was chemotactic for mouse neutrophils, macrophages and lymphocytes. Our results show that mCKLF2, 4, 5 and 6 are four splicing variants which are homologues of human CKLFs and murine CKLFs possess distinct features compared to their human counterparts.

Alternative Splicing↗

Plasma fatty acid composition and incidence of diabetes in middle-aged adults: the Atherosclerosis Risk in Communities (ARIC) Study.

BACKGROUND: The results of some epidemiologic studies conducted by using questionnaires suggest that dietary fat composition influences diabetes risk. Confirmation of this finding with use of a biomarker is warranted. OBJECTIVE: We prospectively investigated the relation of plasma cholesterol ester (CE) and phospholipid (PL) fatty acid composition with the incidence of diabetes mellitus. DESIGN: In 2909 adults aged 45-64 y, plasma fatty acid composition was quantified by using gas-liquid chromatography and was expressed as a percentage of total fatty acids. Incident diabetes (n = 252) was identified during 9 y of follow-up. RESULTS: After adjustment for age, sex, baseline body mass index, waist-to-hip ratio, alcohol intake, cigarette smoking, physical activity, education, and parental history of diabetes, diabetes incidence was significantly and positively associated with the proportions of total saturated fatty acids in plasma CE and PL. The rate ratios of incident diabetes across quintiles of saturated fatty acids were 1.00, 1.36, 1.16, 1.60, and 2.08 (P = 0.0013) in CE and 1.00, 1.75, 1.87, 2.40, and 3.37 (P < 0.0001) in PL. In CE, the incidence of diabetes was also positively associated with the proportions of palmitic (16:0), palmitoleic (16:1n-7), and dihomo-gamma-linolenic (20:3n-6) acids and inversely associated with the proportion of linoleic acid (18:2n-6). In PL, incident diabetes was positively associated with the proportions of 16:0 and stearic acid (18:0). CONCLUSIONS: The proportional saturated fatty acid composition of plasma is positively associated with the development of diabetes. Our findings with the use of this biomarker suggest indirectly that the dietary fat profile, particularly that of saturated fat, may contribute to the etiology of diabetes.

Cholesterol Esters↗

[Effect of N-terminal deletion on biological activity of vascular endothelial cell growth inhibitor].

Vascular endothelial cell growth inhibitor (VEGI) is a novel cytokine which belongs to the TNF superfamily. It can inhibit the proliferation of endothelial cells and neovascularization. However, little is known about the structure-function relationship of VEGI. In order to study the effect of the N-terminus of VEGI on biological properties, the sequence alignment among VEGI and TNF superfamily members based on structure knowledge was done, and then two truncated forms of VEGI were constructed, in which 43 and 51 amino acids from N-terminus were deleted and named VEGI(131) and VEGI(123). Recombinant proteins were generated from E. coli. The expression rates were 25.2% (VEGI(131)) and 27.8% (VEGI(123)) of total bacterial proteins. After purification the purity reached 92.5% (VEGI(131)) and 91.6% (VEGI(123)). VEGI(131) showed significant inhibitory effect on growth of human umbilical vein endothelial cells (HUVEC), IC(50) of VEGI(131) being 35 mg/L. Under the same conditions, IC(50) of VEGI(151) (the wild type of VEGI) was 27 mg/L, but VEGI(123) showed no inhibitory effect. On chick choriallantic membrane (CAM) assay, VEGI(151) markedly reduced the number of main vessels, and VEGI131 decreased capillary number, while the effect of VEGI(123) was almost the same as control. These results suggest that the first 43 amino acids from N-terminus of VEGI have no significant effect on biological activity, but the amino acids 44-51 at N-terminus are required for full biological activity.

Allantois↗

[Purification and stability studies of prokaryotic PDCD5 protein].

OBJECTIVE: To set up an effective and simple purification method to obtain highly purified prokaryotic protein of PDCD5 and study its stability. METHODS: Recombinant PDCD5 protein expressed in E. coli. was accumulated as an inclusion body. After washing, the inclusion body was denatured, renatured, digested with thrombine and then purified by two steps of chromatography. The purity of the products was analyzed by capillary electrophoresis and the stability was identified by SDS-PAGE. RESULTS: Capillary electrophoresis showed that the purity of protein was 100%, and molecular weight was 15,800 with pI 5.9. Further bioactivity assay indicated that the purified PDCD5 could enhance the apoptosis of HL-60 cells withdrawing cytokine, which was in a dose-dependent manner. Stability analysis showed that the PDCD5 protein was sensitive to temperature and easy to degrade at 4 degrees C and 25 degrees C. However, it was relatively stable at -20 degrees C or lyophilized. CONCLUSION: Highly purified and stable recombinant PDCD5 protein was obtained, which lays a foundation for the functional study and application investigation of PDCD5.

Apoptosis Regulatory Proteins↗

Effect of octreotide on angiogenesis induced by hepatocellular carcinoma in vivo.

OBJECTIVE: To investigate the effect of somatostatin analogue octreotide on angiogenesis induced by hepatocellular carcinoma (HCC) in vivo. METHODS: LCI-D20 corneal micropocket model in nude mouse was used to dynamically observe angiogenesis under a stereoscopic zoom microscope and a digital camera system and to evaluate the effect of octreotide on angiogenesis. Male nude mice were subcutaneously implanted with LCI-D20 tumor tissues for tumor xenograft studies. Microvessel density in CD34-stained tumor sections was analyzed by immunohistochemical SP method. RESULTS: Tumor tissues from LCI-D20 implanted into the corneal micropocket induced angiogenesis. When animals received systemic octreotide treatment, angiogenesis response in the cornea of mice was moderate, the appearance of vascular buds was delayed, and the new capillaries were sparse and grew slowly. Compared with the control group, the neovascularization induced by HCC in the cornea of mice was markedly inhibited on day 7, 9, 12, 15, 18 and 21 after implantation in the octreotide-treated group (P<0.05). Systemic administration of octreotide produced a significant suppression of the growth of LCI-D20. Immunohistochemical studies of tumor tissues revealed decreased microvessel density in the octreotide-treated animals as compared with the controls (21.7+/-4.27 versus 31.8+/-3.87, P<0.01). CONCLUSION: Somatostatin analogue octreotide is able to inhibit angiogenesis induced by HCC in vivo and may provide a new approach to the treatment of HCC.

Animals↗

[A novel cis-acting enhancer element between CKLF and CKLFSF1 genes].

AIM: To explore the roles of the CKLF gene and CKLFSF1 gene sequence (CCS) in transcriptional regulation. METHODS: The target gene fragment was amplified by PCR and then inserted into pGL3-basic and pGL3-SV40 containing luciferase reporter vector gene to construct pGL3-basic-CCS and pGL3-SV40-CCS. Using liposome-mediated method, four recombinant plasmids were respectively transfected into Hela cells. Transient expression was analyzed. RESULTS: The luciferase assay indicated that the no luciferase activity was detected in Hela cells transtected with pGL3-basic and pGL3-basic-CCS. However, the luciferase activity was doubled when pGL3-SV40-CCS was transfected into Hela cells. CONCLUTION: The CCS has no promoter activity, whereas some important cis-acting enhancer elements which modulate its downstream gene expression may exist within this sequence.

Base Sequence↗

[Gene expression in rice inflorescence development was monitored by using oligonucleotide microarray].

In this paper we chose 50 rice inflorescence genes from Internet,references. Rice oligonucleotide microarray was prepared by printing the target rice inflorescence genes oligonucleotide. Expression patterns of 50 genes from rice inflorescence in three different development phase were obtained by scanning using ScanArray3000 after array hybridization. The scatter plots and scale maps of the images were acquired after the acquired gene expression patterns were analyzed by ImaGene4.0 software. The scatter plots and scale maps show that there existed a significant difference in the expression of these candidate genes in rice inflorescences with different development phase. Further analysis of those candidate gene expression patterns will be helpful to understand the developmental mechanism of rice inflorescence.

English Abstract↗

Hypohalite ion catalysis of the disproportionation of chlorine dioxide.

The disproportionation of chlorine dioxide in basic solution to give ClO2- and ClO3- is catalyzed by OBr- and OCl-. The reactions have a first-order dependence in both [ClO2] and [OX-] (X = Br, Cl) when the ClO2- concentrations are low. However, the reactions become second-order in [ClO2] with the addition of excess ClO2-, and the observed rates become inversely proportional to [ClO2-]. In the proposed mechanisms, electron transfer from OX- to ClO2(k1OBr- = 2.05 +/- 0.03 M(-1) x s(-1) for OBr(-)/ClO2 and k1OCl-= 0.91 +/- 0.04 M(-1) x s(-1) for OCl-/ClO2) occurs in the first step to give OX and ClO2-. This reversible step (k1OBr-/k(-1)OBr = 1.3 x 10(-7) for OBr-/ClO2, / = 5.1 x 10(-10) for OCl-/ClO2) accounts for the observed suppression by ClO2-. The second step is the reaction between two free radicals (XO and ClO2) to form XOClO2. These rate constants are = 1.0 x 10(8) M(-1) x s(-1) for OBr/ClO2 and = 7 x 10(9) M(-1) x s(-1) for OCl/ClO2. The XOClO2 adduct hydrolyzes rapidly in the basic solution to give ClO3- and to regenerate OX-. The activation parameters for the first step are DeltaH1(++) = 55 +/- 1 kJ x mol(-1), DeltaS1(++) = - 49 +/- 2 J x mol(-1) x K(-1) for the OBr-/ClO2 reaction and DeltaH1(++) = 61 +/- 3 kJ x mol(-1), DeltaS1(++) = - 43 +/- 2 J x mol(-1) x K(-1) for the OCl-/ClO2 reaction.

Journal Article↗

Fibrin fragment D-dimer and the risk of future venous thrombosis.

Plasma D-dimer concentration rises more than 100-fold during acute deep vein thrombosis, but there are no prospective data concerning D-dimer as a risk factor for incident venous thrombosis in a general population. Incident venous thrombosis was ascertained in 2 prospective observational studies, the Atherosclerosis Risk in Communities Study and the Cardiovascular Health Study. Of 21 690 participants enrolled between 1987 and 1993, after 8 years of follow-up, D-dimer was measured using baseline stored plasma of 307 participants who developed venous thrombosis and 616 who did not. Relative to the first quintile of the distribution of D-dimer, the age-adjusted odds ratios for future venous thrombosis for the second to fifth quintiles of D-dimer were 1.6, 2.3, 2.3, and 4.2, respectively (P for trend <.0001). Following added adjustment for sex, race, body mass index, factor V Leiden, prothrombin 20210A, and elevated factor VIII coagulant activity (factor VIII:c), these odds ratios were 1.5, 2.1, 1.9, and 3.0, respectively (P for trend <.0001). Among those with idiopathic thrombosis or secondary thrombosis unrelated to cancer, the adjusted fifth quintile odds ratios were 3.5 and 4.8, respectively. By contrast, D-dimer in the fifth versus first quintile was not related to occurrence of cancer-associated thrombosis (odds ratio, 1.1). Odds ratios for elevated D-dimer were consistently elevated in subgroups defined by age, sex, race, duration of follow-up, and thrombosis type (deep vein thrombosis or pulmonary embolus). D-dimer is strongly and positively related to the occurrence of future venous thrombosis.

Aged↗

Evaluation of deep inspiration breath-hold lung treatment plans with Monte Carlo dose calculation.

PURPOSE: To evaluate dosimetry of deep inspiration breath-hold (DIBH) relative to free breathing (FB) for three-dimensional conformal radiation therapy of lung cancer with 6-MV photons and Monte Carlo (MC) dose calculations. METHODS AND MATERIALS: Static three-dimensional conformal radiation therapy, 6-MV plans, based on DIBH and FB CT images for five non-small-cell lung cancer patients, were generated on a clinical treatment planning system with equivalent path length tissue inhomogeneity correction. Margins of gross to planning target volume were not reduced for DIBH plans. Cord and lung toxicity determined the maximum treatment dose for each plan. Dose distributions were recalculated for the same beams with an MC dose calculation algorithm and electron density distributions derived from the CT images. RESULTS: MC calculations showed decreased target coverage relative to treatment-planning system predictions. Lateral disequilibrium caused more degradation of target coverage for DIBH than for FB (approximately 4% worse than expected for FB vs. 8% for DIBH). However, with DIBH higher treatment doses could be delivered without violating normal tissue constraints, resulting in higher total doses to gross target volume and to >99% of planning target volume. CONCLUSIONS: If DIBH enables prescription dose increases exceeding 10%, MC calculations indicate that, despite lateral disequilibrium, higher doses will be delivered to medium-to-large, partly mediastinal gross target volumes, providing that 6-MV photons are used and margins are not reduced.

Adult↗

High-throughput genotyping of single nucleotide polymorphisms using new biplex invader technology.

The feasibility of large-scale genome-wide association studies of complex human disorders depends on the availability of accurate and efficient genotyping methods for single nucleotide polymorphisms (SNPs). We describe a new platform of the invader assay, a biplex assay, where both alleles are interrogated in a single reaction tube. The assay was evaluated on over 50 different SNPs, with over 20 SNPs genotyped in study cohorts of over 1500 individuals. We assessed the usefulness of the new platform in high-throughput genotyping and compared its accuracy to genotyping results obtained by the traditional monoplex invader assay, TaqMan genotyping and sequencing data. We present representative data for two SNPs in different genes (CD36 and protein tyrosine phosphatase 1beta) from a study cohort comprising over 1500 individuals with high or low-normal blood pressure. In this high-throughput application, the biplex invader assay is very accurate, with an error rate of <0.3% and a failure rate of 1.64%. The set-up of the assay is highly automated, facilitating the processing of large numbers of samples simultaneously. We present new analysis tools for the assignment of genotypes that further improve genotyping success. The biplex invader assay with its automated set-up and analysis offers a new efficient high-throughput genotyping platform that is suitable for association studies in large study cohorts.

Biotechnology↗

Kinetics and mechanisms of the ozone/bromite and ozone/chlorite reactions.

Ozone reactions with XO(2)(-) (X = Cl or Br) are studied by stopped-flow spectroscopy under pseudo-first-order conditions with excess XO(2)(-). The O(3)/XO(2)(-) reactions are first-order in [O(3)] and [XO(2)(-)], with rate constants k(1)(Cl) = 8.2(4) x 10(6) M(-1) s(-1) and k(1)(Br) = 8.9(3) x 10(4) M(-1) s(-1) at 25.0 degrees C and mu = 1.0 M. The proposed rate-determining step is an electron transfer from XO(2)(-) to O(3) to form XO(2) and O(3)(-). Subsequent rapid reactions of O(3)(-) with general acids produce O(2) and OH. The OH radical reacts rapidly with XO(2)(-) to form a second XO(2) and OH(-). In the O(3)/ClO(2)(-) reaction, ClO(2) and ClO(3)(-) are the final products due to competition between the OH/ClO(2)(-) reaction to form ClO(2) and the OH/ClO(2) reaction to form ClO(3)(-). Unlike ClO(2), BrO(2) is not a stable product due to its rapid disproportionation to form BrO(2)(-) and BrO(3)(-). However, kinetic spectra show that small but observable concentrations of BrO(2) form within the dead time of the stopped-flow instrument. Bromine dioxide is a transitory intermediate, and its observed rate of decay is equal to half the rate of the O(3)/BrO(2)(-) reaction. Ion chromatographic analysis shows that O(3) and BrO(2)(-) react in a 1/1 ratio to form BrO(3)(-) as the final product. Variation of k(1)(X) values with temperature gives Delta H(++)(Cl) = 29(2) kJ mol(-1), DeltaS(++)(Cl) = -14.6(7) J mol(-1) K(-1), Delta H(++)(Br) = 54.9(8) kJ mol(-1), and Delta S(++)(Br) = 34(3) J mol(-1) K(-1). The positive Delta S(++)(Br) value is attributed to the loss of coordinated H(2)O from BrO(2)(-) upon formation of an [O(3)BrO(2)(-)](++) activated complex.

Journal Article↗

Protein C, antithrombin, and venous thromboembolism incidence: a prospective population-based study.

Although deficiencies of protein C and antithrombin, 2 natural plasma anticoagulants, are known risk factors for venous thrombosis, population-based prospective incidence data on these associations are lacking. Venous thromboembolic events have been identified in adults in 2 longitudinal cohort studies, the Atherosclerosis Risk in Communities (ARIC) Study and the Cardiovascular Health Study (CHS). Incidence was examined in relation to prediagnostic plasma levels of protein C (ARIC Study only) and antithrombin. Over a mean of 8.1 years of follow-up, there were 130 incident venous thromboembolic events that were not due to cancer in the ARIC Study. The age-adjusted incidence was elevated 3.36-fold (95% CI 1.24 to 9.11) in the 1.1% of subjects with protein C values <2.0 mg/L compared with subjects with higher values. In contrast, in the ARIC Study and the CHS, there was no association between low plasma antithrombin and venous thromboembolism. In conclusion, in this population-based study, a low protein C, but not antithrombin, level has been determined to be associated with an increased incidence of venous thromboembolism. Attributable risk estimates suggest that low protein C levels account for approximately 2.5% of venous thromboembolic events in the ARIC population.

Adult↗

Side-chain lactam-bridge conformational constraints differentiate the activities of salmon and human calcitonins and reveal a new design concept for potent calcitonin analogues.

We have recently reported the potent hypocalcemic effects of side-chain lactam-bridged analogues of human calcitonin (hCT) (Kapurniotu, A.; et al. Eur. J. Biochem. 1999, 265, 606-618). To extend these studies, we have now synthesized a new series of (Asp(17), Lys(21)) and (Asp(17), Orn(21)) side-chain bridged salmon calcitonin (sCT) and hCT analogues. The affinities of these analogues for the human calcitonin receptor, hCTR(I1)(-), and for rat-brain membrane receptors were assayed in competitive binding assays, and agonist potencies at the hCTR(I1)(-) receptors were assessed, using a cAMP-responsive gene-reporter assay. The bridged sCT analogues had activities similar to sCT itself. In contrast, an (Asp(17), Orn(21)) side-chain bridged hCT analogue, cyclo(17-21)-[Nle(8), Phe(12), Asp(17), Orn,(21) Tyr(22))-hCT, was 80 and 450 times more active than hCT in the hCTR(I1)(-) and rat-brain receptor binding assays, respectively, and was 90 times more potent than hCT and 16 times more potent than sCT in initiating receptor signaling. An uncyclized, isosteric analogue of this peptide was also more potent than hCT, demonstrating that the cyclization constraint and these single-residue substitutions enhance the activities of hCT in an additive fashion. This study demonstrates that the potency-enhancing effects of lactam-bridge constraints at hCT residues 17-21 are not transferable to sCT. We also show that, in comparison to the hCT analogues, sCT and its analogues are less potent agonists than expected from their hCTR(I1)(-) affinities. This suggests that it may be possible to preserve the efficient signal transduction of hCT while introducing additional receptor affinity-enhancing elements from sCT into our potent lactam-bridged hCT analogue, thereby creating new super-potent, hCT-based agonists.

Amino Acid Sequence↗