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Biomedical subjects

J Yin

Publications and source records attributed to J Yin.

At least 37 records · Page 2Linked to original sources

Effects of gastric electrical field stimulation with long pulses on gastric emptying in dogs.

The aim was to investigate the effects of electrical field stimulation (EFS) with long and short pulses on gastric emptying, gastric contractility and vagal activity in dogs. Sixteen dogs were equipped with a duodenal cannula, electrodes and strain gauges (10 dogs) in the stomach. Each dog was fed with Ensure and gastric effluent was collected from the cannula. Electrical stimulation was applied via two electrodes (about 12 cm apart, one in the corpus and the other in the antrum) with long pulses (a frequency of 6 cycles min-1, pulse amplitude of 6 mA and width of 100 ms) in 10 dogs and with short pulses (frequency of 30 Hz and pulse width of 300 micros) in six dogs. The electrocardiogram was also recorded and heart rate variability was derived to assess the vagal activity. It was found that: (i). EFS with long pulses did not alter gastric emptying during stimulation but increased gastric emptying during the 45 min immediately after stimulation; (ii). EFS with long pulses increased gastric contractility in both proximal and distal antrum during and after the stimulation; (iii). EFS with long pulses resulted in an increase in vagal tone during the 45 min immediately after stimulation. However, there is no difference during the 45 min period of stimulation; (iv). EFS with short pulses had no effect on gastric emptying. We concluded that long pulse gastric electrical field stimulation with one electrode in the corpus and the other electrode in the antrum has postponed effects on gastric emptying of liquid, gastric contractility and vagal activity.

Animals↗

The effects of obesity on pulmonary function.

AIMS: (1) To determine the predominant pulmonary function abnormality in our population of obese children; and (2) to assess the correlation between the severity of lung function impairment and the degree of obesity as assessed by dual energy x ray absorptiometry (DEXA). METHODS: Sixty four obese patients underwent physical examination, standardised pulmonary function tests (spirometry, lung volumes, and single breath diffusion capacity for carbon monoxide), and DEXA scan measurements. The trunk and subtotal (total - head) body fat mass were used as surrogate index of body adiposity. RESULTS: Sixteen girls and 48 boys with median age and body mass index (BMI) of 12 years (interquartile range (IQR): 10-14) and 30.1 kg/m2 (IQR: 27.2-32.8) respectively were studied. None of the patients had clinical evidence of cardiopulmonary disease. Reduction in functional residual capacity (median FRC 93% predicted, IQR: 68.5-116.5%) and impairment of diffusion capacity (median DLco 83.5% predicted, IQR: 70.0-100.7%) were the most common abnormalities in our cohort, being observed in 30 (46%) and 21 (33%) patients respectively. Obstructive ventilatory impairment was found in three patients. There was significant negative correlation between the degree of reduction of FRC but not DLco with DEXA scan measurements, but such a relation was not found when BMI was used as the indicator of obesity. CONCLUSION: Reduction in FRC and diffusion impairment were the commonest abnormalities found in our cohort of obese patients. Reduction in static lung volume was correlated with the degree of obesity.

Absorptiometry, Photon↗

Sleeping energy expenditure in paediatric patients with obstructive sleep apnoea syndrome.

OBJECTIVE: To investigate sleeping energy expenditure in paediatric patients with obstructive sleep apnoea syndrome. DESIGN: Retrospective study. Setting. University teaching hospital, Hong Kong. PATIENTS AND METHODS: A retrospective analysis comparing sleeping energy expenditure in patients with confirmed obstructive sleep apnoea syndrome and control subjects matched for age, sex, and ideal weight for body height. Subjects were recruited from the Paediatric Chest Clinic and all had undergone overnight polysomnography and sleeping energy expenditure measurement by open-circuit indirect calorimetry using a metabolic cart and canopy system. The measurements were taken during slow-wave sleep. RESULTS: Twenty-four cases with obstructive sleep apnoea syndrome (apnoea hypopnoea index >5) and 23 control subjects were studied. Mean age and ideal weight for body height were 9.4 (standard deviation, 3.9) years and 152.5% (27.2%), respectively. Mean sleeping energy expenditure corrected for body weight for the cases and the control group were 44.83 (standard deviation, 10.49) Kcal/day and 40.71 (10.60) Kcal/day, respectively. Sleeping energy expenditure was not found to be associated with the severity of obstructive sleep apnoea syndrome (r=0.34, P=0.27). CONCLUSION: A trend towards greater sleeping energy expenditure was found in patients with obstructive sleep apnoea syndrome. Further studies on the metabolic aspects of this condition are required.

Calorimetry, Indirect↗

Stochastic vs. deterministic modeling of intracellular viral kinetics.

Within its host cell, a complex coupling of transcription, translation, genome replication, assembly, and virus release processes determines the growth rate of a virus. Mathematical models that account for these processes can provide insights into the understanding as to how the overall growth cycle depends on its constituent reactions. Deterministic models based on ordinary differential equations can capture essential relationships among virus constituents. However, an infection may be initiated by a single virus particle that delivers its genome, a single molecule of DNA or RNA, to its host cell. Under such conditions, a stochastic model that allows for inherent fluctuations in the levels of viral constituents may yield qualitatively different behavior. To compare modeling approaches, we developed a simple model of the intracellular kinetics of a generic virus, which could be implemented deterministically or stochastically. The model accounted for reactions that synthesized and depleted viral nucleic acids and structural proteins. Linear stability analysis of the deterministic model showed the existence of two nodes, one stable and one unstable. Individual stochastic simulation runs could access and remain at the unstable node. In addition, deterministic and averaged stochastic simulations yielded different transient kinetics and different steady-state levels of viral components, particularly for low multiplicities of infection (MOI), where few virus particles initiate the infection. Furthermore, a bimodal population distribution of viral components was observed for low MOI stochastic simulations. The existence of a low-level infected subpopulation of cells, which could act as a viral reservoir, suggested a potential mechanism of viral persistence.

Animals↗

Global gene expression profiling in Barrett's esophagus and esophageal cancer: a comparative analysis using cDNA microarrays.

In order to identify and contrast global gene expression profiles defining the premalignant syndrome, Barrett's esophagus, as well as frank esophageal cancer, we utilized cDNA microarray technology in conjunction with bioinformatics tools. We hybridized microarrays, each containing 8000 cDNA clones, to RNAs extracted from 13 esophageal surgical or endoscopic biopsy specimens (seven Barrett's metaplasias and six esophageal carcinomas). Hierarchical cluster analysis was performed on these results and displayed using a color-coded graphic representation (Treeview). The esophageal samples clustered naturally into two principal groups, each possessing unique global gene expression profiles. After retrieving histologic reports for these tissues, we found that one main cluster contained all seven Barrett's samples, while the remaining principal cluster comprised the six esophageal cancers. The cancers also clustered according to histopathological subtype. Thus, squamous cell carcinomas (SCCAs) constituted one group, adenocarcinomas (ADCAs) clustered separately, and one signet-ring carcinoma was in its own cluster, distinct from the ADCA cluster. We conclude that cDNA microarrays and bioinformatics show promise in the classification of esophageal malignant and premalignant diseases, and that these methods can be applied to small biopsy samples.

Barrett Esophagus↗

Enantiocontrolled synthesis of 2,3,6-trisubstituted piperidines using (eta(3)-dihydropyridinyl)molybdenum complexes as chiral scaffolds. Total synthesis of (-)-indolizidine 209B.

Enantiopure TpMo(CO)2(pyridinyl) complexes were prepared using an efficient and scalable enzymatic kinetic resolution of the precursor to the molybdenum complex. A single TpMo(CO)2(pyridinyl) complex can function as a chiral scaffold for the enantiocontrolled synthesis of either 2,3,6-cis- or 2,6-cis-3-trans-trisubstituted piperidines. The synthetic potential of this methodology was demonstrated by a total synthesis of (-)-indolizidine 209B.

Alkaloids↗

A comparative analysis of the immunological evolution of antibody 28B4.

In an effort to gain greater insight into the evolution of the redox active, catalytic antibody 28B4, the germline genes used by the mouse to generate this antibody were cloned and expressed, and the X-ray crystal structures of the unliganded and hapten-bound germline Fab of antibody 28B4 were determined. Comparison with the previously determined structures of the unliganded and hapten-bound affinity-matured Fab [Hsieh-Wilson, L. C., Schultz, P. G., and Stevens, R. C. (1996) Proc. Natl. Acad. Sci. U.S.A. 93, 5363] shows that the germline antibody binds the p-nitrophenyl ring of hapten 3 in an orientation significantly different from that seen in the affinity-matured antibody, whereas the phosphonate moiety is bound in a similar mode by both antibodies. The affinity-matured antibody 28B4 has more electrostatic and hydrophobic interactions with hapten 3 than the germline antibody and binds the hapten in a lock-and-key fashion. In contrast, significant conformational changes occur in the loops of CDR H3 and CDR L1 upon hapten binding to the germline antibody, consistent with the notion of structural plasticity in the germline antibody-combining site [Wedemayer, G. J., Patten, P. A., Wang, L. H., Schultz, P. G., and Stevens, R. C. (1997) Science 276, 1665]. The structural differences are reflected in the differential binding affinities of the germline Fab (K(d) = 25 microM) and 28B4 Fab (K(d) = 37 nM) to hapten 3. Nine replacement mutations were found to accumulate in the affinity-matured antibody 28B4 compared to its germline precursor. The effects of each mutation on the binding affinity of the antibody to hapten 3 were characterized in detail in the contexts of both the germline and the affinity-matured antibodies. One of the mutations, Asp95(H)Trp, leads to a change in the orientation of the bound hapten, and its presence is a prerequisite for other somatic mutations to enhance the binding affinity of the germline antibody for hapten 3. Thus, the germline antibody of 28B4 acquired functionally important mutations in a stepwise manner, which fits into a multicycle mutation, affinity selection, and clonal expansion model for germline antibody evolution. Two other antibodies, 20-1 and NZA6, with very different antigen specificities were found to be highly homologous to the germline antibody of 28B4, consistent with the notion that certain germline variable-region gene combinations can give rise to polyspecific hapten binding sites [Romesberg, F. E., Spiller, B., Schultz, P. G., and Stevens, R. C. (1998) Science 279, 1929]. The ultimate specificity of the polyspecific germline antibody appears to be defined by CDR H3 variability and subsequent somatic mutation. Insights into the evolution of antibody-combining sites provided by this and other structural studies are discussed.

Amino Acid Sequence↗

Instabilotyping: comprehensive identification of frameshift mutations caused by coding region microsatellite instability.

Coding region frameshift mutation caused by microsatellite instability (MSI) is one mechanism contributing to tumorigenesis in cancers with MSI in high frequency. Mutation of TGFBR2 is one example of this process. To identify additional examples, a large-scale genomic screen of coding region microsatellites was conducted. 1115 coding homopolymeric loci with six or more nucleotides were identified in an online genetic database. Mutational screening was performed at 152 of these loci in 46 colorectal tumors with MSI in high frequency. Nine loci were mutated in > or =20% of tumors, 10 loci in 10-20%, 24 loci in 5-10%, 43 loci in <5%, and 66 loci were not mutated in any tumors. The most frequently mutated novel loci were the activin type II receptor gene (58.1%), SEC63 (48.8%), AIM 2 (47.6%), a gene encoding a subunit of the NADH-ubiquinone oxidoreductase complex (27.9%), a homologue of mouse cordon-bleu (23.8%), and EBP1/PA2G4 (20.9%). This genome-wide approach identifies coding region MSI in genes or pathways not implicated previously in colorectal tumorigenesis, which may merit functional study or other additional analysis.

3' Untranslated Regions↗

Hypermethylation of the hMLH1 gene promoter is associated with microsatellite instability in early human gastric neoplasia.

A significant portion of gastric cancers exhibit defective DNA mismatch repair, manifested as microsatellite instability (MSI). High-frequency MSI (MSI-H) is associated with hypermethylation of the human mut-L homologue 1 (hMLH1) mismatch repair gene promoter and diminished hMLH1 expression in advanced gastric cancers. However, the relationship between MSI and hMLH1 hypermethylation has not been studied in early gastric neoplasms. We therefore investigated hMLH1 hypermethylation, hMLH1 expression and MSI in a group of early gastric cancers and gastric adenomas. Sixty-four early gastric neoplasms were evaluated, comprising 28 adenomas, 18 mucosal carcinomas, and 18 carcinomas with superficial submucosal invasion but clear margins. MSI was evaluated using multiplex fluorescent PCR to amplify loci D2S123, D5S346, D17S250, BAT 25 and BAT 26. Methylation-specific PCR was performed to determine the methylation status of hMLH1. In two hypermethylated MSI-H cancers, hMLH1 protein expression was also evaluated by immunohistochemistry. Six of sixty-four early gastric lesions were MSI-H, comprising 1 adenoma, 4 mucosal carcinomas, and 1 carcinoma with superficial submucosal invasion. Two lesions (one adenoma and one mucosal carcinoma) demonstrated low-frequency MSI (MSI-L). The remaining 56 neoplasms were MSI-stable (MSI-S). Six of six MSI-H, one of two MSI-L, and none of thirty MSI-S lesions showed hMLH1 hypermethylation (P<0.001). Diminished hMLH1 protein expression was demonstrated by immunohistochemistry in two of two MSI-H hypermethylated lesions. hMLH1 promoter hypermethylation is significantly associated with MSI and diminished hMLH1 expression in early gastric neoplasms. MSI and hypermethylation-associated inactivation of hMLH1 are more prevalent in early gastric cancers than in gastric adenomas. Thus, hypermethylation-associated inactivation of the hMLH1 gene can occur early in gastric carcinogenesis.

Adaptor Proteins, Signal Transducing↗

Oxygen consumption by lungs with acute and chronic injury in a rabbit model.

OBJECTIVE: To study the oxygen consumption by lungs with acquired acute and chronic injury in a rabbit model. DESIGN: A non-randomized controlled animal study. SUBJECTS AND METHOD: Three groups of White New Zealand rabbits were studied to determine the oxygen consumption of the lungs. Group 1 (n=21, controls) consisted of healthy rabbits with normal lungs. The rabbits in group 2 (n=14) had sustained acute lung damage induced by subcutaneous N-nitroso-N-methylurethane and those in group 3 (n=9) had sustained chronic lung damage inflicted by a single dose of intra-tracheal bleomycin. Pulmonary oxygen consumption was estimated from the difference between the whole body oxygen consumption, measured by indirect calorimetry, and systemic oxygen consumption estimated by the Fick method. MEASUREMENTS AND RESULTS: Both acute and chronic lung-damaged groups had significantly greater pulmonary oxygen consumption than the control group, both as absolute values [control vs acute vs chronic: 0.25 (0.04) vs 0.76 (0.10) vs 1.77 (0.21) ml/kg per min, p<0.001], and as a percentage of total body oxygen consumption [2.98 (0.47) vs 9.34 (1.39) vs 20.20 (2.11)%, p<0.0001]. Histological evaluation of the severity of lung damage using a lung-injury score revealed significantly higher scores in both lung-injury groups than in the controls. CONCLUSIONS: These findings suggest that the lung is an important site of energy loss in subjects with acute and chronic lung injury.

Acute Disease↗

A comparison of cytokine responses in respiratory syncytial virus and influenza A infections in infants.

UNLABELLED: Respiratory syncytial virus (RSV) infection is a major cause of bronchiolitis in infants while influenza A infection usually manifests as upper respiratory tract infection. We hypothesised that the immunological responses of infants to RSV infection and influenza A infection are different. This prospective study was undertaken to compare the cytokine responses during RSV and influenza A infection. Sera and nasopharyngeal aspirates (NPA) were collected from infants with a coryzal illness with or without wheeze who were admitted to the paediatric wards during 1998. Cytokines, adhesion molecules, RANTES, IgE and eosinophil cationic protein (ECP) were measured by enzyme linked immunosorbent assay or fluorescence enzyme immunoassay. The diagnosis of RSV and influenza infections was based on direct immunofluorescence and viral culture. Of the 39 infants studied, RSV infection was confirmed in 11 patients and Influenza A in 10 patients. All RSV patients and one influenza A patient had wheeze during the infection. The serum concentrations of interleukin (IL)-4 and IL-5, regulated upon activation normal T cell expressed and secreted (RANTES) and soluble intercellular adhesion molecule 1 (sICAM-1) in infants with RSV infection were significantly higher than those with influenza A infection (all P < 0.02). The concentration of tumour necrosis factor-alpha (TNF-alpha) in NPA was significantly lower in infants with RSV infection (P < 0.01). CONCLUSION: A predominant T helper cell type 2 cytokine and related immunological response was observed in infants with respiratory syncytial virus infection whereas a predominant pro-inflammatory cytokine response was observed in infants with influenza A infection. This may explain the different clinical manifestations of the two viral infections in infants.

Cell Adhesion Molecules↗

A study of a new TSM bio-mimetic sensor using a molecularly imprinted polymer coating and its application for the determination of nicotine in human serum and urine.

A new bio-mimetic quartz crystal thickness-shear-mode (TSM) sensor, using an imprinted polymer coating as the sensitive material, has been fabricated and applied to the determination of nicotine (NIC) in human serum and urine. The molecularly imprinted polymer (MIP) was synthesized using NIC as the template molecule and methacrylic acid (MAA) as the functional monomer. The sensor showed high selectivity and a sensitive response to NIC in aqueous system. The linear response range of the sensor was between 5.0 x 10(-8) and 1.0 x 10(-4) M with a detection limit of 2.5 x 10(-8) M. The viscoelasticity of the coating in the air and in liquid has been studied by the impedance spectrum. The MIP sensor was stable and exhibited effective reproducibility. Satisfactory results were achieved in the detection of the real samples.

Biosensing Techniques↗

Quantitative and qualitative assessment of articular cartilage in the goat knee with magnetization transfer imaging.

We investigated the role of collagen in the magnetization transfer (MT) effect in contrast to other macromolecules. By means of phantoms made of collagen, chondroitin sulfate (CS) and albumin, MR parameters have been optimized in order to reduce the acquisition time and improve the sensitivity, as well as to minimize the contributions from CS and albumin to the MT induced signal attenuation. The same method was used to study cartilage ex vivo (bovine articular and nasal cartilage plugs) and in vivo (goat knee femoral chondyle). In phantom samples, the MT signal attenuation depended on the collagen concentration while contributions from the other macromolecules were found to be minimal. In average, analysis of MT images revealed a approximately 25%, approximately 35% and approximately 30% signal attenuation in 10% w/v type I collagen gels, cartilage plugs, and cartilage from the weight-bearing areas of the goat knee, respectively. Biochemical data revealed that treatment of cartilage plugs with bacterial collagenase led to collagen depletion and correspondingly to a decrease of the MT response. In contrast, trypsin-induced proteoglycan loss in cartilage plugs did not alter the MT effect. A significant correlation was observed between the collagen content in these plugs and their respective MT ratios and the rate constant k for the exchange process bound versus free water. Finally, data obtained from in vivo MT measurement of the goat knee demonstrated that intra-articular injection of papain might not only cause degradation of proteoglycans but also a change in collagen integrity in a dose-dependent manner. We conclude that in vivo measurement of MT ratios gives quantitative and qualitative information on the collagen status and may be applied for the routine evaluation of normal and abnormal articular cartilage.

Animals↗

Simulating the growth of viruses.

To explore how the genome of an organism defines its growth, we have developed a computer simulation for the intracellular growth of phage T7 on its E. coli host. Our simulation, which incorporates 30 years of genetic, biochemical, physiological, and biophysical data, is used here to study how the intracellular resources of the host, determined by the specific growth rate of the host, contribute toward phage development. It is also used to probe how changes in the linear organization of genetic elements on the T7 genome can affect T7 development. Further, we show how time-series trajectories of T7 mRNA and protein levels generated by the simulation may be used as raw data to test data-mining strategies, specifically, to identify partners in protein-protein interactions. Finally, we suggest how generalization of this work can lead to a knowledge-driven simulation for the growth of any virus.

Algorithms↗

[Purification and biological activity of rh-leptin expressed in Escherichia coli].

The human leptin was successfully expressed with high level in E. coli under the control of PL promotor. The yield of recombinant protein was over 40% of total cellular protein and expressed as inclusion bodies. The recombinant human leptin (rh-leptin) was purified with gel filtration, anion-exchange and reverse chromatography. Refolding was achieved by gradually reducing denaturant using a diafiltration method. The refolded rh-leptin was characterized by SDS-PAGE, Western-blotting and its first 15 amino acid residues sequence of the N-terminal. The purified product was found to be biologically active, reducing the food intake and body weight gain upon testing in BALB/c mice.

Animals↗

[Retention of selenium volatility using lime in coal combustion].

For understanding the volatility of selenium, the effect of the contents of exchangeable cations of coal on it, and the retention of selenium using CaO in coal combustion, the sequential chemistry extraction, the fixed bed and circulating fluidized bed (CFB) combustion, X-ray diffraction (XRD) and atomic fluorescence spectrometry (AFS) were undertaken. The results showed that the volatility of selenium was more than 97% in coal combustion at 815 degrees C, and the volatility of selenium was affected by the content of exchangeable cations of coal in low-middle temperature. It was identified that lime can restrain the volatility of selenium. In fixed bed combustion of coal, the retention rates of selenium volatility were between 11.6% and 50.7% using lime. In circulating fluidized bed combustion of coal, partitioning of selenium changed very much in ash of different size fraction between without lime and with lime. Comparing with combustion without lime, the content of selenium in ash from chimney was less than fourth times and that in leaching water from chimney decreased by two orders of magnitude using lime. Retention of selenium volatility using lime is so effective in coal combustion, especially in CFB combustion of coal.

Calcium Compounds↗

[Clone and expression of human soluble CD14 and study of its function].

Human soluble CD14(sCD14) cDNA fragment was amplified using total RNA extracted from U937 cells by RT-PCR of sCD14 gene, and the recombinant expression plasmid pEF1/HisC/sCD14 348aa was constructed. Then the expression in eukaryotic cell was carry out by liposome transfection method. It demonstrated that the expression level was relatively high by scanning map identification. The expressed product was purified by immunoaffinity chromatography and the purity was above 90%. The changes of CD14 brought by LPS stimulating U937 cell proved the product had the function of combine with LPS.

Cloning, Molecular↗