Biomedical subjects

Y T Wu

Publications and source records attributed to Y T Wu.

At least 19 recordsLinked to original sources

PubMed2006

High performance liquid chromatographic determination of a new antifungal compound, ADKZ in rat plasma.

A high performance liquid chromatography (HPLC) method was developed and validated for the determination of ADKZ (1-(1H-1,2,4-triazole)-2-(2,4-diflurophenyl) -3-[N-methyl-N-(4-iodo-benzyl)amino]-2-propanol) in rat plasma. The compound was extracted from plasma samples by liquid-liquid extraction, and an isomeric compound of ADKZ (1-(1H-1,2,4-triazole)-2-(2,4-diflurophenyl)-3-[N-methyl-N -(3-iodo-benzyl)amino]-2-propanol) was used as the internal standard (IS), which were analyzed on a reversed-phase C18 column (5 microm, 200 mm x 4.6 mm i.d.). The extracted plasma samples were eluted with acetonitrile-0.018 M triethylamine solution adjusted to pH 3.2 with phosphoric acid (35:65, v/v). The effluent was monitored by a UV detector at 230 nm. The retention time of ADKZ was 7.1 min and IS 8.2 min. The calibration curves were linear in the concentration range of 0.02-2.00 microg/ml with the correlation coefficients greater than 0.999. The quantification limit of ADKZ in rat plasma was 0.02 microg/ml. Intra- and inter-day precision ranged from 2.6 to 7.9% and 3.1 to 9.6%, respectively. The extraction recovery from plasma was no less than 80%. No endogenous interferences were observed with either ADKZ or IS. The method has been successfully used to support the pre-clinical pharmacokinetic studies of ADKZ in rats.

Acetonitriles↗

PubMed2005

Chiral liquid chromatography resolution and stereoselective pharmacokinetic study of tetrahydropalmatine enantiomers in dogs.

A selective chiral high performance liquid chromatographic (HPLC) method coupled with achiral column was developed and validated to separate and quantify tetrahydropalmatine (THP) enantiomers in dog plasma. Chromatography was accomplished by two steps: (1) racemic THP was separated from biological matrix and collected on a Kromasil C18 column (150 mmx4.6 mm, 5 microm) with the mobile phase acetonitrile-0.1% phosphoric acid solution, adjusted with triethylamine to pH 6.15 (47:53); (2) enantiomeric separation was performed on a Chiralcel OJ-H column (250 mmx4.6 mm, 5 microm) with the mobile phase anhydrous ethanol. The detection wavelength was set at 230 nm. (+)-THP and (-)-THP were separated with a resolution factor (Rs) of at least 1.6 and a separation factor (alpha) greater than 1.29. Linear calibration curves were obtained over the range of 0.025-4 microg/ml in plasma for each of (+)-THP and (-)-THP (R2>0.999) with a limit of detection (LOD) of 0.005 microg/ml and the recovery was greater than 88% for each enantiomer. The relative standard deviation (R.S.D.) and relative error values were less than 10% at upper and lower concentrations. The method was used to determine the pharmacokinetics of THP enantiomers after oral administration of racemic THP. The results presented herein showed the stereoselective disposition kinetics of THP in dogs and were a further contribution to the understanding of the kinetic behavior of THP analogues.

Animals↗

PubMed2004

Study of stereoselective pharmacokinetics of anisodamine enantiomers in rabbits by capillary electrophoresis.

The purpose of this study was to determine the pharmacokinetics of anisodamine enantiomers in plasma after oral and intravenous administration of racemic anisodamine in rabbits. A capillary electrophoresis method for the simultaneous separation of two pairs of enantiomers in plasma has been firstly developed and validated. Using a 75 mM phosphate buffer containing 25 mM carboxymethylated-gamma-cyclodextrin at pH 2.5, good resolution was achieved on a 45-cm uncoated fused-silica capillary at the voltage of 20 kV and 25 degrees C. The pharmacokinetics of individual anisodamine enantiomers were characterized using the CE assay, the sole method of enantiomeric separation for anisodamine. Pharmacokinetic analysis of results indicated that anisodamine enantiomers showed non-stereoselective disposition or stereoselective disposition in different rabbits. For the rabbits with non-stereoselective disposition, similar pharmacokinetic characteristics were observed between (6S, 2'S)- and (6R, 2'R)-, or (6S, 2'R)- and (6R, 2'S)-anisodamine. For the rabbits with stereoselective disposition, (6S, 2'S)- and (6R, 2'S)-anisodamine were below the established LOD, while the two remaining enantiomers also had similar pharmacokinetic profiles. Further investigations remain necessary to find out the underlying mechanism about the stereoselective disposition of (6S, 2'S)- and (6R, 2'S)-anisodamine.

Animals↗

PubMed2004

Alteration of the copy number and deletion of mitochondrial DNA in human hepatocellular carcinoma.

Somatic mutations in mitochondrial DNA (mtDNA) have been detected in hepatocellular carcinoma (HCC). However, it remains unclear whether mtDNA copy number and mitochondrial biogenesis are altered in HCC. In this study, we found that mtDNA copy number and the content of mitochondrial respiratory proteins were reduced in HCCs as compared with the corresponding non-tumorous livers. MtDNA copy number was significantly reduced in female HCC but not in male HCC. Expression of the peroxisome proliferator-activated receptor gamma coactivator-1 was significantly repressed in HCCs (P<0.005), while the expression of the mitochondrial single-strand DNA-binding protein was upregulated, indicating that the regulation of mitochondria biogenesis is disturbed in HCC. Moreover, 22% of HCCs carried a somatic mutation in the mtDNA D-loop region. The non-tumorous liver of the HCC patients with a long-term alcohol-drinking history contained reduced mtDNA copy number (P<0.05) and higher level of the 4977 bp-deleted mtDNA (P<0.05) as compared with non-alcohol patients. Our results suggest that reduced mtDNA copy number, impaired mitochondrial biogenesis and somatic mutations in mtDNA are important events during carcinogenesis of HCC, and the differential alterations in mtDNA of male and female HCC may contribute to the differences in the clinical manifestation between female and male HCC patients.

Alcohol Drinking↗

PubMed2004

Genomic sequence of an isolate of Newcastle disease virus isolated from an outbreak in geese: a novel six nucleotide insertion in the non-coding region of the nucleoprotein gene. Brief Report.

We have completely sequenced the genome of an isolate of Newcastle disease virus (NDV) obtained following an outbreak in geese. The genomic sequence consists of 15192 nt, which is six nt longer than the published full length genome of the NDV strains La Sota, B1, Clone-30, Beaudette C and HB V4. The six nt insertion was located in the non-coding region of the nucleoprotein (NP) gene between nt 1646 and nt 1647 of the NDV genome (numbered according to the genomic sequence of the La Sota strain). An additional 22 NDV strains were searched for the existence of this six nt insertion. NDVs in genotypes VI, VII, VIII and IX had this insertion while NDV's in genotypes I, II, III, IV, and V did not. The significance of this insertion in NDV evolution is discussed.

Animals↗

PubMed2003

Pathotypical and genotypical characterization of strains of Newcastle disease virus isolated from outbreaks in chicken and goose flocks in some regions of China during 1985-2001.

Twenty-nine strains of Newcastle disease virus (NDV) isolated from outbreaks in chicken and goose flocks in several regions of China during 1985-2001 were characterized pathotypically and genotypically. All except one of these strains were velogenic. For genotyping, phylogenetic tree analysis (nt 47-420), restriction site mapping (nt 334-1682) and residue substitution analysis (residues 4-124) of the F gene were performed using sequences of our isolates and sequences of selected NDV strains from GenBank. The results revealed that most of these newly characterized strains belonged to six novel genetic groups that were designated as VIf, VIg, VIIc, VIId, VIIe and IX. The genotype IX viruses, to which the China challenge strain F48E8 used for vaccine evaluation belonged, were found only in China and still induced sporadic infections in certain areas. Isolates belonging to group VIf and VIg were distinct from previously reported members of genotype VI, with genetic distance from 2.5 to 12.1%. Subgenotype VIIc, VIId and VIIe viruses, which were distributed in clusters in the phylogenetic tree distinct from members of subgenotypes VIIa and VIIb, were responsible for disease outbreaks in chicken and goose flocks and circulated predominantly in southern China in recent years. Finally, cross-protective testing showed that specific-pathogen free (SPF) chickens vaccinated with La Sota vaccines can be fully protected against challenge by strains from genetic groups VIb, VIg, VIId and IX, indicating that the antigenic differences between strains of various genotypes are insufficient to change the cross-protection conferred by the commonly used vaccine.

Amino Acid Sequence↗

PubMed2001

A left-lateralized network for reading Chinese words: a 3 T fMRI study.

fMRI was used to investigate brain organization for reading in Chinese. Subjects were shown two-character Chinese words. A control task was used to eliminate the non-linguistic visual and motor confounds. Results show that naming of Chinese logographs is characterized by left-lateralized neuronal networks for the processing of orthographic, phonological, and semantic attributes. The orchestration of the middle frontal cortex, superior temporal cortex, superior parietal cortex, basal temporal area and extrastriate cortices of the left hemisphere may manifest the particularity of the central representation of simple word naming in Chinese.

Adult↗

PubMed2001

Separation of endo-polygalacturonase using aqueous two-phase partitioning.

The partitioning of endo-polygalacturonase (endo-PG) in polyethylene glycol (PEG)-polyvinyl alcohol (PVA10000) and PEG-hydroxypropyl starch (Reppal PES100) aqueous two-phase systems was studied, and revealed the possibility of using aqueous two-phase extraction to purify and concentrate endo-PG from its clarified fermentation broth. For the PEG8000-PVA10000 system, endo-PG presented in the fermentation broth (at concentration that is more than 40% of total protein) mainly dominates in the top phase with a partitioning coefficient of 6, while total protein concentrates in the bottom phase. A separation scheme consisting of two consecutive aqueous two-phase extraction steps was proposed: a first extraction in polyethylene glycol (PEG8000)-polyvinyl alcohol system, followed by a second extraction in PEG8000-(NH4)2SO4 system. This allowed the separation of endo-PG from polymer and the recycling of PEG polymer, since endo-PG was very strongly partitioned into the bottom phase of the PEG8000-(NH4)2SO4 system. Laboratory-scale experiments were performed to test the efficiency of this scheme. It was found that enzyme recovery was up to 91% with a total purification factor of about 1.9 and a concentration factor of more than 5. About 90% of the total PEG added into the systems can be recovered, and no reduction was obtained in the purification factor using recycled PEG.

Culture Media↗

PubMed2001

Proteomic analysis of proteins in PC12 cells before and after treatment with nerve growth factor: increased levels of a 43-kDa chromogranin B-derived fragment during neuronal differentiation.

Proteomic analysis is an important approach to characterizing the proteome and studying protein function in the post-genomic era. It is also a powerful screening method for detecting unexpected alterations in protein expression that may be missed by conventional biochemical techniques. The aim of this study was to perform a preliminary proteomic analysis of PC12 cells in order to investigate the effect of nerve growth factor (NGF) on protein expression in PC12 cells during neurite outgrowth. PC12 cell proteins were separated by two-dimensional electrophoresis (2DE) and visualized by silver staining, then certain proteins were identified by N-terminal amino acid microsequencing and a homology search of a protein sequence database. Over 400 proteins were detected, 10% of which showed a significant (greater than 30%) increase or decrease in expression during NGF-induced neuronal differentiation. Seven proteins in the 2DE map were identified; the levels of five of these were unaffected by NGF treatment, whereas the levels of the other two, beta-tubulin and a novel 43-kDa chromogranin B-derived fragment, were significantly increased by more than 30 and 200%, respectively. Our results suggest that chromogranin B processing is enhanced in PC12 cells during NGF-induced neuronal differentiation. In addition, since this increase in the levels of the chromogranin B-derived fragment was specifically blocked by PD98059, we suggest that the increased processing can be ascribed to activation of the MAP kinase pathway, and that the 43-kDa chromogranin B-derived fragment can serve as a new marker of neuronal differentiation for proteomic studies.

Animals↗

PubMed2001

Activation of the hypothalamus characterizes the acupuncture stimulation at the analgesic point in human: a positron emission tomography study.

We performed a positron emission tomography study, using regional cerebral blood flow as the index of brain activity, to address the specificity of brain activation pattern by acupuncture stimulation of short duration at the classical analgesic point. Needling manipulation at 2 Hz was performed at a classical point of prominent analgesic efficacy (Li 4, Heku) and a near-by non-classical/non-analgesic point, respectively, in normal subjects. Regions activated by acupuncture stimulation at Li 4 included the hypothalamus with an extension to midbrain, the insula, the anterior cingulate cortex, and the cerebellum. Of note, it was only the stimulation at Li 4 that activated the hypothalamus under the similar psychophysical ratings of acupuncture sensation (deqi) as elicited by the stimulation at the two points, respectively. The data suggested that the hypothalamus might characterize the central expression of acupuncture stimulation at the classical analgesic point and serve as one key element in mediating analgesic efficacy of acupuncture stimulation.

Acupuncture Analgesia↗

PubMed2001

Structural modeling and characterization of a thermostable lipase from Bacillus stearothermophilus P1.

The moderate thermophilic bacterium Bacillus stearothermophilus P1 expresses a thermostable lipase that was active and stable at the high temperature. Based on secondary structure predictions and secondary structure-driven multiple sequence alignment with the homologous lipases of known three-dimensional (3-D) structure, we constructed the 3-D structure model of this enzyme and the model reveals the topological organization of the fold, corroborating our predictions. We hypothesized for this enzyme the alpha/beta-hydrolase fold typical of several lipases and identified Ser-113, Asp-317, and His-358 as the putative members of the catalytic triad that are located close to each other at hydrogen bond distances. In addition, the strongly inhibited enzyme by 10 mM PMSF and 1-hexadecanesulfonyl chloride was indicated that it contains a serine residue which plays a key role in the catalytic mechanism. It was also confirmed by site-directed mutagenesis that mutated Ser-113, Asp-317, and His-358 to Ala and the activity of the mutant enzyme was drastically reduced.

Amino Acid Sequence↗

PubMed2001

Comparisons of cutaneous blood flow reactivity to norepinephrine and sodium nitroprusside between patients with heart transplantation and healthy subjects.

Heart transplant patients are reported to have impaired regulation of the microvasculature. The purpose of this study was to investigate the cutaneous blood flow and its reactivity to sodium nitroprusside (vasodilator, Nipride 0.1%) and norepinephrine (vasoconstrictor, Levophed 0.1%) in patients after heart transplantation in comparison to normal healthy individuals. Eighteen patients after heart transplantation and 16 healthy, nonsmoking individuals served as subjects of the study. Sodium nitroprusside and norepinephrine were introduced by iontophoresis to the skin of the right and left forearms, respectively. After measuring cutaneous blood flow reactivity in the pre-exercise state by laser Doppler flowmetry, subjects were then asked to close and open their fists for 2 min. The same measurements were repeated after exercise. Comparisons between the groups were carried out by the Wilcoxan signed rank test. The Mann Whitney U-test was used for comparison between pre-exercise and post-exercise states. The results demonstrated that sodium nitroprusside significantly increased forearm cutaneous perfusion at rest but produced only a mild increase after exercise. This reactivity was significantly lower after exercise with no significant differences between groups. Norepinephrine decreased cutaneous blood flow at rest. The transplant patients were significantly less sensitive to norepinephrine before but not after exercise. The changes in norepinephrine reactivity with exercise were significantly different between groups (p < 0.05).

Exercise↗

PubMed2001

The relationship between exercise performance and peripheral circulation in patients with peripheral arterial occlusive disease.

The exercise performance of patients with peripheral arterial occlusive disease was often impaired. The relationship of exercise performance, daily physical activity, and peripheral circulation was studied. Twenty-five patients with peripheral arterial occlusive disease were recruited; their exercise performance was evaluated with a maximal graded exercise test. A habitual physical activity questionnaire was used to quantify the physical activity level. Measurement of peripheral circulation included the assessment of macrocirculation (ankle-brachial pressure index, calf blood flow by impedance plethysmograph) and microcirculation (cutaneous blood flow, cutaneous temperature, transcutaneous oxygen tension, and cutaneous blood flow responses to iontophoresis). Pearson's product correlation coefficient and multiple linear regression were used in data analysis. Results showed that age, sports index in habitual physical activity questionnaire, and ankle-brachial pressure index were significantly correlated with maximal walking time. Age and sports index were the two major determinants for their exercise performance and accounted for a 61.5% variability.

Arterial Occlusive Diseases↗

PubMed2001

Retrograde transcatheter occlusion of patent ductus arteriosus: preliminary experience in Gianturco coil technique without heparinization.

BACKGROUND: The aim of this study was to report the initial experience of using the Gianturco coil (Cook Cardiology, Bloomington, Indiana) without heparinization to close patent ductus arteriosus (PDA). PATIENTS AND METHODS: Forty consecutive patients (30 females, 10 males) underwent transcatheter closure of a PDA via the right femoral artery approach without heparinization. Patients ages ranged from 7 months to 55 years (median, 6.8 years); weights ranged from 7.8 65 kg (median, 18.3 kg). Twenty-one patients had cardiomegaly (n = 21), congestive heart failure (n = 10), or both (n = 10). The PDAs measured 0.8 4.5 mm (median, 2.6 mm) at the narrowest diameter; the mean Qp/Qs ranged from 1.0 2.6 (1.4 +/- 0.4). The helical diameter of the coil that we chose was 1.7 times the narrowest PDA diameter. The length of the coil was sufficient to produce 4 or 5 loops. RESULTS: Successful coil placement was accomplished in all 40 patients. Thirty-five patients (87.5%) underwent single coil implantation, 2 patients (5%) had 2 coils, and 3 patients (7.5%) had 3 coils. Complete ductus occlusion was achieved in 80% of cases at the end of the procedure, while 8 patients had minimal shunt detected by aortography or echocardiography. The occlusion rate increased to 87.5% by the next day, 90% by 1 month, 92.5% by 3 months, 95% by 6 months, and 97.5% by 9 and 12 months. Only one asymptomatic patient had minimal residual shunt detected by color Doppler at 12 months follow-up. At a median follow-up of 17 months (range, 5 25 months), no patient had thromboembolism, endocarditis, coil migration, diminished femoral pulse or hemolysis. CONCLUSION: Our preliminary results suggest that retrograde transcatheter closure of PDA with the Gianturco coil without heparinization is feasible, safe and efficacious. A single coil allowed complete occlusion of PDA 3 mm in diameter.

Adolescent↗

PubMed2001

[Genetic diversity detected by DNA markers and phenotypes in Upland cotton].

Genetic diversity of Upland cotton (Gossypium hirsutum L.) cultivars was evaluated by SSR, ISSR, and RAPD. The genomic DNAs of 36 domestic and exotic cultivars were amplified with 81 SSR primer pairs; 7 ISSR primers and 53 RAPD primers, which yielded 282 polymorphic bands. The range of pair similarity coefficient (Nei, M. And W. Li 1979) between cultivars is from 0.5745 to 0.9291. The result indicated that although the present cultivars have genetic variation to a certain extent, the genetic basis of them is relatively narrow, and it is essential to find some new germplasm for the development of new cultivars. As expected, the classification of the cotton cultivars using the Unweighted Pair-group Method with Arithmetic Average based on the pair similarity coefficients is mostly consistent with known pedigree information and breeding experience. The 36 cotton cultivars have been mainly classed into exotic cultivar group, Chinese Xinjiang ecological region cultivar group, early-mature domestic cutivar group and mediate-mature domestic cultivar group. Carmen and Colorado cultivars from Spain have the lowest similarity with other cultivars, while TM-1 (genetic standard line), MD5lne and PD9364 cultivars from American have higher similarity with domestic cultivars. The reason may be that the parents of the domestic cultivars were mainly introduced from America. However, clusters of domestic cultivars are not all in accordance with their ecological regions, which may result from the pedigree complexity of the cultivars. Meanwhile the genetic diversity of 36 cultivars was investigated at phenotypic level with agronomic and fiber quality traits in the field experiments in 1999 and 2000, and was measured by Euclidean distance on the genotypes. Although grouping result of cluster analysis based on the Euclidean distance is similar than that of molecular data with a significant negative correlation of -0.3350 between the pair similarity coefficients and the Euclidean distance, the subgroup classification of the two methods gives different result. The molecular markers can be effectively applied in classifying cultivars within the group. The study provides useful information for the selection of parents in the development of cotton cultivars and hybrids as well.

Cluster Analysis↗

PubMed2001

[SFE-CGC determination of tanshinone in Salvia miltiorrhiza Bunge].

AIM: To analyze tanshinone IIA in Salvia miltiorrhiza Bunge by supercritical fluid extraction (SFE) coupled with capillary gas chromatography (CGC). METHODS: The solution pressure was predicted by solubility parameter theory, the extraction conditions were confirmed by orthogonal design in which temperature, volume of modifier and dynamic extracting volume had been studied in three levels. RESULTS: This method is quick, convenient, precise and accurate, added sample recovery is 95.3%, RSD = 4.27%. CONCLUSION: Compared with the method of ultrasonic extraction, significantly better qualitative and quantitative result could be obtained by SFE-CGC and expend less organic solvent then make less pollution.

Abietanes↗

PubMed2000

Efficacy of a 24-week aerobic exercise program for osteopenic postmenopausal women.

Osteoporosis is one of the most common skeletal disorders affecting postmenopausal women. The purpose of this study was to investigate whether a 24-week program of aerobic high-impact loading exercise was beneficial for enhancing physical fitness and bone mineral density (BMD) in osteopenic postmenopausal women. Forty-three postmenopausal women aged 48-65 years participated in this study. The BMD of the spine (L2-L4) and right femoral neck of each woman was below 1 SD of the mean of premenopausal women, as examined by dual X-ray absorptiometry. The assignment of subjects into exercise or control group was not randomized but based on each subject's anticipated compliance to the 6-month long exercise program. Twenty-two subjects joined the exercise group and attended the training programs and 21 served as the control group. Exercise programs included treadmill walking at an intensity above 70% of maximal oxygen consumption (VO2max) for 30 minutes, followed by 10 minutes of stepping exercise using a 20-cm-high bench. The program was conducted three times per week for 24 weeks. Physical fitness measurements included testing of flexibility, muscular strength and endurance, body composition, and cardiopulmonary fitness. The results showed that the quadriceps strength, muscular endurance, and VO2max in the exercise group had significant improvements, whereas no improvement was found in any of the physical fitness parameters in the control group. The BMD of the L2-L4 and the femoral neck in the exercise group increased 2.0% (P > 0.05) and 6.8% (P < 0.05) and those in the control group decreased 2.3% (P < 0.05) and 1.5% (P > 0.05), respectively. In conclusion, aerobics combined with high-impact exercise at a moderate intensity was effective in offsetting the decline in BMD in osteopenic postmenopausal women.

Absorptiometry, Photon↗

PubMed2000

Targeting delivery of paclitaxel into tumor cells via somatostatin receptor endocytosis.

BACKGROUND: The binding of somatostatin (SST) to endogenous G-protein-coupled receptors (SST receptors or SSTRs) is followed by internalization of SST, and, several reports have shown that a high density of SSTRs is present on most hormone-secreting tissue tumors. Facile synthesis of the long-acting SST analog, octreotide, has previously been described. Octreotide might be of practical value in developing tumor tracers and in serving as a carrier of cytotoxic antitumor drugs. RESULTS: Fluorescein-labeled octreotide was internalized into the cytosol of human breast MCF-7 carcinoma cells via binding to SSTRs. Octreotide-conjugated paclitaxel (taxol) was created by coupling taxol-succinate to the amino-terminal end of octreotide. This conjugate retains the biological activity of taxol in inducing formation of tubulin bundles, eventually causing apoptosis of MCF-7 cells. Cytotoxicity of octreotide-conjugated taxol is mainly mediated by SSTR, as shown by the observation that octreotide pretreatment can rescue the induced cell death. In comparison with free taxol, this conjugate shows much less toxicity in Chinese hamster ovary cells. CONCLUSIONS: Octreotide-conjugated taxol exerts the same antitumor effect of free taxol on stabilizing microtubule formation and inducing cell death. This conjugate triggers tumor cell apoptosis mediated by SSTRs and is exclusively toxic to SSTR-expressing cells. Octreotide-conjugated taxol is less toxic to low-SSTR-expressing cells compared with free taxol. Our results strongly indicated that octreotide-conjugated taxol demonstrates cell selectivity and may be used as a targeting agent for cancer therapy.

Animals↗