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Biomedical subjects

W Wang

Publications and source records attributed to W Wang.

At least 955 records · Page 53Linked to original sources

In vitro induction of proinflammatory cytokine secretion by juvenile rheumatoid arthritis synovial fluid immune complexes.

OBJECTIVE: To characterize juvenile rheumatoid arthritis synovial fluid (SF) immune complexes and to examine their interaction with leukocytes. METHODS: SF immunoglobulin-containing fractions were prepared by sequential chromatography on protein A and Sephacryl 300. Fractions were subdivided according to molecular weight, characterized for immunoglobulin and complement content, and incubated with either promonocytic U937 cells or normal human peripheral blood mononuclear cells (PBMC). RESULTS: High molecular weight SF immunoglobulin-containing fractions stimulated the release of interleukin-1beta (IL-1beta) from U937 cells. These same complexes stimulated tumor necrosis factor alpha (TNFalpha), IL-1beta, IL-6, IL-8, and granulocyte-macrophage colony-stimulating factor (GM-CSF) from PBMC. Lower molecular weight material was less efficient in inducing any of the cytokines. TNFalpha and IL-1beta were the earliest of the messenger RNAs examined to be induced by the high molecular weight complexes. However, the secretion of IL-6, IL-8, and GM-CSF stimulated by the complexes was not completely dependent upon the secretion of IL-1beta. Addition of IL-1 receptor antagonist to the cell cultures reduced GM-CSF and IL-6 production by 40% and IL-8 production by 25% in PBMC. CONCLUSION: SF immunoglobulin fractions contain immune complexes that vary in size, composition, and phlogistic potential. High molecular weight complexes are capable of inducing a spectrum of proinflammatory cytokines, all of which have been implicated in the pathogenesis of rheumatic disease.

Adolescent↗

Distribution of lutein and zeaxanthin stereoisomers in the human retina.

The distribution of macular pigment stereoisomers in the human retina has been mapped and a pathway to account for the presence of the non-dietary carotenoid, meso-zeaxanthin, is proposed. Adult neural retinas were cut into three concentric areas centered on the fovea, and the extracted carotenoids were analysed and purified by high-performance liquid chromatography. The dicarbamate or dibenzoate derivatives of the collected zeaxanthin fractions for each tissue sample were further analysed by HPLC to determine their stereoisomer composition. Whole retinas from infant eyes were similarly analysed. The results show that, relative to zeaxanthin, the concentration of lutein in the adult neural retina increases with radial distance from the fovea while that of meso-zeaxanthin decreases. Infant retinas were found to have more lutein and less meso-zeaxanthin, relative to zeaxanthin, than adult retinas. Small quantities of (3S, 3'S)-zeaxanthin were also found in the adult retina, particularly in the macula. It is proposed that lutein and zeaxanthin are transported into an individual's retina in the same proportions found in his or her blood serum. Some of the lutein is then converted into meso-zeaxanthin, primarily in the macula, by a mechanism which is less developed in infants than adults.

Age Factors↗

Right ventricular collagen type III and IV gene expression increases during early phases of endurance training in hypobaric hypoxic condition.

The objective of this study was to examine the effects of prolonged exposure to hypobaric hypoxic condition, physical training and their combination on collagen type I, III and IV gene expression in the ventricles and atria of rat heart. Male rats were assigned to four groups: normobaric sedentary (NS) and trained (NT), and hypobaric sedentary (HS) and trained (HT). Exposure to and treadmill running training in hypobaric condition were carried out in a hypobaric chamber (770-740 mbar, 2250-2550 m). Experimental periods were 10, 21 and 56 days; the groups of 91 days served as recovery groups from experimental settings of 56 days. Exposure to hypobaric condition as such and in combination with endurance training for 10 days increased right ventricular weight-to-body weight ratio (RV/BW) by 26% (p < 0.001) and 23% (p < 0.01), respectively, when compared to 10NS. RV/BW was significantly increased also in 21HT and 56HT. Left ventricular weight-to-body weight ratio was 13% (p < 0.01) and 14% (p < 0.01) higher in 21HT and 56HT, respectively, than in the respective NS. Right ventricular collagen type III mRNA level was 33% (p = 0.065) and 38% (p < 0.01) higher in 10HT than in 10NS and 10NT, respectively. Right ventricular collagen type IV mRNA level was 29% (p < 0.001) higher in 10HT than in 10NS. Relatively slight left ventricular hypertrophy was not associated with significant changes in collagen mRNA levels. Decreased left ventricular subepicardial prolyl 4-hydroxylase activity in 10HS and 10HT suggests transient corresponding decrease in the rate of collagen synthesis. This study shows that combination of endurance training and moderate hypobaric hypoxic condition leads to increased right ventricular collagen type III and IV gene expression associated with right ventricular hypertrophy.

Animals↗

Prospective study of early detection for primary liver cancer.

PURPOSE: To determine whether repeated screening can lead to early detection of primary liver cancer (PLC) and in turn to an improved clinical result. METHODS: In this randomized controlled study, Shanghai urban residents aged 35-55 years and with serum evidence of HBV infection or chronic liver disease were eligible for recruitment. Using cluster sampling, these subjects were allocated into two groups-the screening group and the control group: there were 8109 subjects in the screening group and 9711 in the control group. Subjects in the screening group were tested with serum AFP and real-time ultrasound every 6 months. One to four rounds of screening were completed. Liver cancer was treated according to stage at diagnosis. RESULTS: All subjects enrolled were followed up and classed at the end-point as alive without liver cancer, alive with liver cancer, dead from liver cancer, or dead from another cause. The mean follow-up was 1.2 years; total follow-up was 12,038 person-years in the screening group and 9,573 person-years in the control group. We detected 38 patients with PLC in the screening group and 18 patients with PLC in the control group. In the patients in the screening group 76.8% of patients were at a subclinical stage, and 70.6% of them underwent resection, the 1- and 2-year survival rates being 88.1% and 77.5%, respectively. However, in the control group, none of the patients was at a subclinical stage when diagnosed, none of them underwent resection, and none of them survived over 1 year. The lead time was estimated at 0.45 years. The cost of detecting PLC at an early stage was RMB 12,600 (US$1,500). CONCLUSION: The study proved that screening the high-risk population for PLC with a serum AFP test and real-time ultrasound examination can detect patients in the early stages, increase the resection rate and prolong the survival time. It is therefore recommended that screening for PLC be advocated in any high-risk area.

Adult↗

Effects of avitriptan, a new 5-HT 1B/1D receptor agonist, in experimental models predictive of antimigraine activity and coronary side-effect potential.

Several acutely acting antimigraine drugs, including ergotamine and sumatriptan, have the ability to constrict porcine arteriovenous anastomoses as well as the human isolated coronary artery. These two experimental models seem to serve as indicators, respectively, for the therapeutic and coronary side-effect potential of the compounds. Using these two models, we have now investigated the effects of avitriptan (BMS-180048; 3-[3-[4-(5-methoxy-4-pyrimidinyl)-l-piperazinyl[propyl]-N-methyl-l H-indole-5-methane-sulfonamide monofumarate), a new 5-HT 1B/1D receptor agonist. In anaesthetized pigs, avitriptan (10, 30, 100 and 300 micrograms.kg-1) decreased the total carotid blood flow by exclusively decreasing arteriovenous anastomotic blood flow; capillary blood flow was increased. The mean +/- SEM i.v. dose of avitriptan eliciting a 50% decrease (ED50) in the porcine carotid arteriovenous anastomotic blood flow was calculated to be 76 +/- 23 micrograms.kg-1 (132 +/- 40 nmol.kg-1) and the highest dose (300 micrograms.kg-1) produced a 72 +/- 4% reduction. In recent comparative experiments (DeVries et al. 1996), the mean +/- SEM ED50 (i.v.) of sumatriptan in decreasing carotid arteriovenous anastomotic blood flow was 63 +/- 17 micrograms.kg-1 (158 +/- 43 nmol.kg-1), with a reduction of 76 +/- 4% by 300 micrograms.kg-1, i.v. Both avitriptan (pD2; 7.39 +/- 0.09; Emax: 13.0 +/- 4.5% of the contraction to 100 mM K+) and sumatriptan (pD2: 6.33 +/- 0.09; Emax: 15.5 +/- 2.3% of the contraction to 100 mM K+) contracted the human isolated coronary artery. The above results suggest that avitriptan should be able to abort migraine headaches in patients, but may exhibit sumatriptan-like effects on coronary arteries. Initial clinical studies have demonstrated the therapeutic action of the drug in acute migraine.

Animals↗

The effect of different sampling intervals on the measurement of intrapartum fetal heart rate variability.

OBJECTIVE: To test the hypothesis that increasing the sampling interval affects the intrapartum fetal heart rate (FHR) variability measurement. METHODS: Fetal electrocardiograms were obtained from women in labor. Using the peak of the fetal R wave, the R-R interval and FHR were calculated on a beat-to-beat basis. Retrospectively, the original data were repartitioned using different intervals (2-900 seconds) to generate a window of measurement (epoch). The mean value for each epoch and the last FHR in that epoch (epochal value) were compared with published animal and human data. Errors were quantified by comparing the epochal and mean values for each epoch. Fetal heart rate variability between epochs and within each epoch was compared. RESULTS: Fetal heart rate and R-R interval were measured in 146 cases. The FHR had a normal distribution (mean 140.1 beats per minute, +/- standard deviation [SD] 15.6, skew -0.07), but its inverse, the R-R interval, was not normally distributed (mean 432 milliseconds, +/- SD 52.4, skew 1.78). Using a single value for an epoch duration of 2 seconds resulted in an error that was similar to the within-epoch variability (+/- SD of 2.2 beats per minute difference between mean and epochal value compared to +/- SD of 2 beats per minute within epoch) but which increased with epoch duration. CONCLUSION: An epoch duration of 2 seconds and a single sampled value within this period may be appropriate for measurement of both medium and long-term variability in any computerized intrapartum FHR interpretation system. Fetal heart rate (not R-R interval, because of its normal distribution) should be used to design such a computerized system.

Adult↗

Evaluation of the biocompatibility of a nonceramic hydroxyapatite.

In this study, the biocompatibility of calcium phosphate cement (CPC) was systematically investigated, which included systemic injection acute toxicity assay, cell culture cytotoxicity assay, gene mutation assay (Ames test), chromosome aberration assay (micronucleus test), DNA damage assay (unscheduled DNA synthesis test) and implant histological evaluation. The results showed that the CPC had no toxicity and all tests for mutagenicity and potential carcinogenicity of CPC extracts are negative. CPC was implanted into the femurs of rabbits under no-load condition and the histology of specimens revealed that the implant tightly joined with the surrounding bone, only a very slight histological inflammatory reaction occurred. Thus, CPC is a highly biocompatible material and seems to be safe for application in humans.

Animals↗

A simple and efficient method for purification of prawn baculovirus DNA.

A new method for isolation of prawn baculovirus and subsequent extraction of viral DNA was developed. No density gradient centrifugation, ultracentrifugation or phenol-chloroform extraction steps were involved. Phenylmethylsulfonyl fluoride (PMSF) was used to prevent proteinase degradation, DNase and RNase were used to degrade prawn DNA and RNA respectively. The nucleocapsid was a bacilliform virion, about 58 62 nm in width and 300-350 nm in length as observed by transmission electron microscopy. Intact viral DNA was obtained by lysing nucleocapsids with guanidine hydrochloride and degrading protein with proteinase K. As the viral DNA was digested with restriction endonuclease and separated by electrophoresis, restriction fragments were clearly shown on the agarose gel. The size of the DNA was estimated approximately to be 290 kb. The virus which appeared to be a prawn baculovirus was named prawn white spot baculovirus (PWSBV) due to the white spots which appeared on the inside surface of the crust of infected prawns.

Animals↗

Effects of neuronal proteoglycans on activity-dependent growth responses of fetal hippocampal neurons.

Excitatory amino-acid (EAA) neurotransmitters act as molecular signals influencing the structure of neurons during development. However, the signal transduction and effector mechanisms responsible for these effects have yet to be fully elucidated. We have previously provided evidence that EAA agonists induce the synthesis and release of proteoglycans (PGs) with neurite-promoting activity from fetal hippocampal neurons. In the present studies exposure of fetal hippocampal neurons to glutamate (100 microM) for 5 min resulted in increases in the neuron-specific growth-associated genes T alpha 1 alpha-tubulin (T alpha 1), microtubule-associated protein-2 (MAP-2) and growth-associated protein-43 (GAP-43). mRNA levels peaked at between 8 and 12 h following exposure as determined by competitive reverse transcription polymerase chain reaction (RT-PCR). Increases in neurite growth as measured by axonal length, the total length of dendrites, the number of branches per axon, the total length of branches per axon and the total neurite length were also observed 48 h after glutamate exposure. The increase in T alpha 1, MAP-2 and GAP-43 mRNA levels following glutamate exposure was mediated via both N-methyl-D-aspartate and metabotropic receptor activation. Heparin, which inhibits the neurite growth-promoting effects of PGs in vitro, and heparitinase, which catalyzes the cleavage of heparan sulphate, also inhibited the glutamate-dependent induction of T alpha 1, MAP-2 and GAP-43 mRNA expression and neurite growth when added to culture medium following glutamate exposure. Chondroitin sulphate and chondroitinase AC had no effects on the mRNA levels tested or on neurite growth. Therefore, these studies suggest that neuronal PGs regulated by activation of EAA receptors mediate neuronal growth responses.

Animals↗

Expression of rat pro cholecystokinin (CCK) in bacteria and in insect cells infected with recombinant baculovirus.

Neuropeptide prohormones are generally not abundant in nature as they exits to be processed and contain sites which could be cleaved by a number of cellular proteases. In order to study the processing of prohormones in vitro it is necessary to produce them in quantity. Finding an expression system which produces intact prohormone has been a matter of trial and error. We report that intact rat pro CCK was produced with an amino-terminal His-Tag in e. coli, and it was secreted from sf9 and other insect cells infected with a recombinant baculovirus vector. The bacteria contained about 0.1 micrograms pro CCK/ml of cells. The High 5 insect cells produced 4.3 micrograms/ml medium (as determined by RIA), 10 times as much as sf9 or sf21. Using a combination of ion exchange, gel filtration and HPLC, the insect cell protein was purified about 150 fold with a recovery of about 16%. The secreted insect cell pro CCK is tyrosine sulfated like its mammalian equivalent. Using these expression systems it is possible to produce significant (microgram to mg) quantities of pro CCK for immunologic, enzymatic and structural studies.

Animals↗

Quantitative acoustic characterization of a new surfactant-based ultrasound contrast agent.

The acoustic properties of a new ultrasound contrast agent, ST68, have been investigated. ST68 is a sonicated mixture of nonionic surfactants (Span-type and Tween-type) consisting of stabilized microbubbles with a mean diameter of 3.8 microns and a concentrations of 7.1 x 10(8) bubbles/mL. A pulsatile flow system was used to acquire data in vitro. The acoustic properties of ST68, as a function of time, frequency and dose, were calculated. Enhancement changed nonlinearly with contrast agent dose; maximum was 13.1 dB +/- 1.0 dB for a dose of 0.30 microL/mL of suspending medium. Attenuation reached approximately 11 dB/cm for dosages above 0.27 microL/mL and for frequencies between 2.5 and 6.0 MHz. In vivo, i.v. injections of ST68 were given to 4 rabbits (doses from 0.01 to 0.23 mL/kg). A clear increase in flow signal intensity was observed for 1 to 2 min. An in vivo dose-response curve was calculated from audio Doppler signals obtained with a 10-MHz cuff transducer placed around the distal aorta. Maximum enhancement was 18.3 dB +/- 3.13 dB for a 0.13 mL/kg dose. Moreover, ST68 appears to follow a simple relationship between in vivo enhancement and dose. In conclusion, ST68 is capable of producing marked vascular enhancement. Its acoustic properties have been characterized in vitro and in vivo.

Acoustics↗

Influence of an antiprogestin (onapristone) on in vivo and in vitro fertilization.

The effects of a progesterone antagonist (onapristone) on heat synchronization, luteinizing hormone (LH) surge, ovulation, oocyte maturation and fertilization of superovulated ewes were studied. Its effects on in vitro bovine oocyte maturation and fertilization were also studied. Estrus synchronization and superovulation treatments were applied to 39 adult ewes using an intravaginal sponge with fluorgestone acetate for 9 days with injections of prostaglandin F2 alpha and pregnant mare's serum gonadotrophin given 24 h before sponge withdrawal. The animals were randomly assigned to four different groups; T1 receiving only the synchrony treatment (n = 11); T2 ewes received two injections of onapristone (1 mg kg-1, i.v.) 12 h apart from 3 h after sponge withdrawal (n = 10); T3 ewes received two injections of progesterone 12 h apart from sponge withdrawal (n = 10); and, T4 ewes received both onapristone and progesterone as described (n = 8). Ewes were mated by a fertile male during estrus. Progesterone and LH were measured during the superovulation period in plasma samples taken every 4 h. Uterine flushings for ova recovery were performed at 5 days (n = 25), 48 h (n = 5) and 24 h (n = 5). Non-fertilized oocytes collected at 24 and 48 h were checked for meiosis resumption. The effects of two doses of onapristone (D1 and D2) on in vitro bovine oocyte maturation (control = 100, D1 = 100 and D2 = 100) and fertilization (control = 107, D1 = 40 and D2 = 75) were also studied. The percentage of animals showing heat signs was significantly lower in group T3 (50% vs. 100%). The onset of oestrus (27.6, 24.8, 68.8 and 25.5 h, respectively for T1, T2, T3 and T4) and an LH surge (32.3, 28.8, 76.5 and 30.5 h, respectively for T1, T2, T3 and T4) after sponge withdrawal were significantly delayed in group T3. There were no significant differences in the intervals between estrus and LH surge among groups (4.61 +/- 0.75 h). The response and ovulation rates until 40 h after sponge withdrawal (group T3 excluded) were similar among groups, but the fertilization rates were significantly lower in groups T2 and T4 when compared with T1 (2% and 3% vs. 41%, respectively; P < 0.001) due to sperm arrest in the cervix. Ova recovery rate decreased significantly from 24-48 h to 5 days and was not affected by treatments (76.9% vs. 37.1% respectively). Onapristone did not affect the resumption of meiosis. Fertilization of bovine oocytes in vitro decreased significantly only in group D2 when compared to control (48% vs. 62.6%, respectively). In conclusion, onapristone treatment during the preovulatory period did not interfere with normal synchronization of estrus, ovulation and oocyte maturation but severely compromised fertilization by arresting spermatozoa in the cervix.

Animals↗

The HLA-A*0201-restricted H-Y antigen contains a posttranslationally modified cysteine that significantly affects T cell recognition.

A peptide recognized by two cytotoxic T cell clones specific for the human minor histocompatibility antigen H-Y and restricted by HLA-A*0201 was identified. This peptide originates from SMCY, as do two other H-Y epitopes, supporting the importance of this protein as a major source of H-Y determinants in mice and humans. In naturally processed peptides, T cells only recognize posttranslationally altered forms of this peptide that have undergone modification of a cysteine residue in the seventh position. One of these modifications involves attachment of a second cysteine residue via a disulfide bond. This modification has profound effects on T cell recognition and also occurs in other class I MHC-associated peptides, supporting its general importance as an immunological determinant.

Animals↗

c-Rel is a target of pentoxifylline-mediated inhibition of T lymphocyte activation.

The possible clinical use of the methyl xanthine derivative, pentoxifylline (PF), for the treatment of T cell-dependent diseases is being noted with increasing interest. In this paper, we studied the molecular consequences of PF treatment during lymphocyte activation. We found that in T cells, anti-CD3-induced c-Rel expression was blocked by PF, whereas the induction of other NF-kappaB family members was not significantly affected. However, induction of NF-AT, which has the same signaling requirements as c-Rel induction, was not inhibited by PF. Among genes that respond to these transcription factors, IL-2 mRNA induction was suppressed by PF, whereas IL-2R(alpha) chain mRNA induction was not affected. These observations implicated c-Rel as an IL-2 promoter factor, for which experimental support was obtained from transient transfection experiments. In contrast with the observation in T cells, c-Rel induction was not blocked by PF in B cells. The greater selectivity of PF, compared with FK506, at both the molecular and cellular levels may prove advantageous in manipulating T cell responses in vivo.

Animals↗