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Biomedical subjects

Y Wei

Publications and source records attributed to Y Wei.

At least 73 records · Page 4Linked to original sources

Carbon monoxide produced by isolated arterioles attenuates pressure-induced vasoconstriction.

Studies were conducted on isolated rat gracilis muscle arterioles to examine the role of vascular heme oxygenase (HO)-derived carbon monoxide (CO) on myogenic constrictor responses to stepwise increments in intraluminal pressure. The arterioles express HO-2 but not HO-1 and manufacture CO. Both HO-2 protein expression and CO production are reduced in arterioles maintained for 18 h before experimentation in media containing HO-2 antisense oligodeoxynucleotides (AS-ODN). Pressurization of arterioles mounted on a myograph over the pressure range of 40--100 mmHg elicits reduction of internal diameter. At pressures >40 mmHg, the internal diameter of vessels treated with either HO-2 AS-ODN, the HO inhibitor chromium mesoporphyrin (CrMP), or the K(+) channel blocker tetraethylammonium (TEA) are smaller than the corresponding control values. The inclusion of exogenous CO, but not of biliverdin, in the superfusion buffer attenuates pressure-induced vasoconstriction in CrMP-treated vessels. However, exogenous CO does not attenuate pressure-induced vasoconstriction in vessels treated with both CrMP and TEA. Collectively, these data suggest that CO of vascular origin attenuates pressure-induced arteriolar constriction via a mechanism involving a TEA-sensitive K(+) channel.

Animals↗

Inherent capacity for lipogenesis or dietary fat retention is not increased in obesity-prone rats.

Obesity results from positive energy balance and, perhaps, abnormalities in lipid and glycogen metabolism. The purpose of this study was to determine whether differences in lipogenesis, retention of dietary fat, and/or glycogenesis influenced susceptibility to dietary obesity. After 1 wk of free access to a high-fat diet (HFD; 45% fat by energy) rats were separated on the basis of 1 wk body weight gain into obesity-prone (OP; > or =48 g) or obesity-resistant groups (OR; < or =40 g). Rats were either studied at this time (OR1, OP1) or continued on the HFD for an additional 4 wk (OR5, OP5). Weight gain and energy intake were greater (P < or = 0.05) in OP vs. OR at both 1 (53 +/- 2 vs. 34 +/- 1 g; 892 +/- 27 vs. 755 +/- 14 kcal) and 5 (208 +/- 7 vs. 170 +/- 7 g; 4,484 +/- 82 vs. 4,008 +/- 72 kcal) wk, respectively. Rats were injected with (3)H(2)O and were either provided free access to an HFD meal containing labeled fatty acids (fed; n = 10 or 11/group) or were fasted (n = 10/group) overnight. The amount of food or (14)C tracer eaten overnight was equivalent between OP and OR rats. In liver, the fraction of (3)H retained in glycogen or lipid was not significantly different between OR and OP groups. Retention of dietary fat in the liver was not increased in OP rats. In adipose tissue, retention of (3)H was approximately 49% greater (P < or = 0.05) in OP1 vs. OR1 and approximately 30% greater in OP5 vs. OR5, but retention of dietary fat was not elevated in OP vs. OR. At the same time, fat pad weight (sum of epididymal, retroperitoneal, mesenteric) was 49% greater in OP1 rats vs. OR1 rats and 65% greater in OP5 vs. OR5 rats (P < or = 0.05). Thus a greater capacity for lipogenesis or retention of dietary fat does not appear to be included in the OP phenotype. The characteristic increase in energy intake associated with OP rats appears to be necessary and critical to accelerated weight and fat gain.

Adipose Tissue↗

Role of 20-HETE in mediating the effect of dietary K intake on the apical K channels in the mTAL.

We have used the patch-clamp technique to study the effect of dietary K intake on the apical K channels in the medullary thick ascending limb (mTAL) of rat kidneys. The channel activity, defined by the number of channels in a patch and the open probability (NPo), of the 30- and 70-pS K channels, was 0.18 and 0.11, respectively, in the mTAL from rats on a K-deficient diet. In contrast, NPo of the 30- and 70-pS K channels increased to 0.60 and 0.80, respectively, in the tubules from animals on a high-K diet. The concentration of 20-hydroxyeicosatetraenoic acid (20-HETE) measured with gas chromatography-mass spectrometry was 0.8 pg/microg protein in the mTAL from rats on a high-K diet and increased significantly to 4.6 pg/microg protein in the tubules from rats on a K-deficient diet. Addition of N-methylsulfonyl-12,12-dibromododec-11-enamide (DDMS) or 17-octadecynoic acid (17-ODYA), agents that inhibit the formation of 20-HETE, had no significant effect on the activity of the 30-pS K channels. However, DDMS/17-ODYA significantly increased the activity of the apical 70-pS K channel from 0.11 to 0.91 in the mTAL from rats on a K-deficient diet. In contrast, inhibition of the cytochrome P-450 metabolism of arachidonic acid increased NPo from 0.64 to 0.81 in the tubules from animals on a high-K diet. Furthermore, the sensitivity of the 70-pS K channel to 20-HETE was the same between rats on a high-K diet and on a K-deficient diet. Finally, the pretreatment of the tubules with DDMS increased NPo of the 70-pS K channels in the mTAL from rats on a K-deficient diet to 0.76. We conclude that an increase in 20-HETE production is involved in reducing the activity of the apical 70-pS K channels in the mTAL from rats on a K-deficient diet.

Amides↗

Effect of dietary K intake on apical small-conductance K channel in CCD: role of protein tyrosine kinase.

We have used Western blot to examine the expression of cSrc protein tyrosine kinase (PTK) and protein tyrosine phosphatase (PTP)-1D in the renal cortex, and the patch-clamp technique to determine the role of PTK in mediating the effect of dietary K intake on the small-conductance K (SK) channel in the cortical collecting duct (CCD). When rats were on a K-deficient (KD) diet for 1, 3, 5, and 7 days, the expression of cSrc increased by 40, 90, 140, and 135%, respectively. In contrast, the expression of cSrc in the renal cortex from rats on a high-K (HK) diet for 1, 2, and 3 days decreased by 40, 60, and 75%, respectively. However, the protein level of PTP-1D was not significantly changed by dietary K intake. The addition of 1 microM herbimycin A increased NP(o), a product of channel number (N) and open probability (P(o)) in the CCD from rats on a normal diet or on a KD diet. The increase in NP(o) was 0.30 (normal), 0.45 (1-day KD), 0.65 (3-day KD), 1.55 (5-day KD), and 1.85 (7-day KD), respectively. Treatment of the CCD with herbimycin A from rats on a KD diet increased NP(o) per patch from the control value (0.7) to 1.4 (1-day KD), 1.6 (3-day KD), 2.6 (5-day KD), and 3.5 (7-day KD), respectively. In contrast, HK intake for as short as 1 day abolished the effect of herbimycin A. Furthermore, the expression of ROMK channels in the renal cortex was the same between rats on a KD diet or on a HK diet. Moreover, treatment with herbimycin A did not further increase NP(o) in the CCDs from rats on a HK diet. We conclude that dietary K intake plays a key role in regulating the activity of the SK channels and that PTK is involved in mediating the effect of the K intake on channel activity in the CCD.

Animals↗

PKA-induced stimulation of ROMK1 channel activity is governed by both tethering and non-tethering domains of an A kinase anchor protein.

We have used the patch-clamp technique to explore the role of A Kinase Anchor Proteins (AKAP) in mediating the effect of cAMP on ROMK1 channels expressed in the Xenopus oocytes. Addition of membrane permeant cAMP analogs increased channel activity only in oocytes injected with ROMK1 and AKAP79 cRNA but had no effect on channel activity in oocytes injected with ROMK1 alone. Using the two-electrode voltage clamp technique, we determined that application of H89, a potent inhibitor of protein kinase A (PKA), abolished the stimulatory effect of cAMP/forskolin. To investigate the role of AKAP specificity in conferring cAMP responses to ROMK1 channels, we examined channel activity in oocytes expressing ROMK1 and either AKAP18, AKAP-KL or AKAP75. Addition of cAMP failed to increase channel current in oocytes expressing ROMK1 and either AKAP18 or AKAP-KL. In contrast, cAMP increased ROMK1 channel activity by 33% in oocytes coexpressing AKAP75, the bovine homologue of AKAP79. The effect of cAMP on ROMK1 in oocytes coexpressing AKAP75 is inhibited by H89. Since all three AKAPs bind PKAII, the results suggest that a unique structural domain in AKAP75/79 collaborates with the PKAII binding site and enables a productive association of PKA with ROMK1 channels. Deletion of either the membrane targeting region of AKAP75 (AKAP45) or PKAII binding domain of AKAP75 (AKAP75DeltaC) abolished the effects of forskolin on ROMK1 channels. This suggests that the membrane targeting and the PKA binding domains of AKAP75 are essential for the effect of cAMP. However, the nature of the AKAP, that interacts with ROMK1 in the native tissue, remains to be determined because AKAP75/79 are not expressed in the kidney. We conclude that the regulation of ROMK1 channels by PKA requires the involvement of the cell membrane-directed AKAPs that are able to specifically link PKA to the target channel protein.

Animals↗

A novel method for enzyme immobilization: direct encapsulation of acid phosphatase in nanoporous silica host materials.

Immobilization of acid phosphatase (ACP) in mesoporous or, more generally, nanoporous silica has been accomplished via the sol-gel reactions of tetramethyl orthosilicate in the presence of ACP and of D-glucose (DG) as a nonsurfactant template, which is subsequently removed by water extraction after the formation of nanocomposite gels. Characterization of the silica host after the removal of DG shows that the pore size and volume generally increase with the DG content. At high DG contents, the silica hosts are nanoporous with interconnected nanoscaled pores/channels of regular diameter (e.g., 3.4 nm). Catalytic activity of ACP encapsulated in nanoporous hosts is significantly improved over that in microporous host prepared in the absence of DG. The apparent enzymatic activity at various pH values and substrate concentrations correlates well with the nanostructures of the host matrices. As the DG content is increased in the synthesis, the activity tends to increase. At a DG content of 42-60 wt%, the samples exhibit activities about triple that of the template-free control. These and other results from enzymatic kinetic studies suggest that the increase in the pore size and volume facilitates the transport of the substrate and product molecules in the host matrices, leading to the observed increase in activity. The thermal stability of ACP is remarkably improved upon immobilization. There is no detectable leakage of ACP from the host matrices and the biogels are reuseable. This study provides a useful protocol for the development of nanotechnology for various biocatalysts and biosensors.

Acid Phosphatase↗

Carbon monoxide of vascular origin attenuates the sensitivity of renal arterial vessels to vasoconstrictors.

Rat renal interlobar arteries express heme oxygenase 2 (HO-2) and manufacture carbon monoxide (CO), which is released into the headspace gas. CO release falls to 30% and 54% of control, respectively, after inhibition of HO activity with chromium mesoporphyrin (CrMP) or of HO-2 expression with antisense oligodeoxynucleotides (HO-2 AS-ODN). Patch-clamp studies revealed that CrMP decreases the open probability of a tetraethylammonium-sensitive (TEA-sensitive) 105 pS K channel in interlobar artery smooth muscle cells, and that this effect of CrMP is reversed by CO. Assessment of phenylephrine-induced tension development revealed reduction of the EC(50) in vessels treated with HO-2 AS-ODN, CrMP, or TEA. Exogenous CO greatly minimized the sensitizing effect on agonist-induced contractions of agents that decrease vascular CO production, but not the sensitizing effect of K channel blockade with TEA. Collectively, these data suggest that vascular CO serves as an inhibitory modulator of vascular reactivity to vasoconstrictors via a mechanism that involves a TEA-sensitive K channel.

Animals↗

Highly sensitive determination method for total carbonate in water samples by flow injection analysis coupled with gas-diffusion separation.

A spectrophotometric method for the determination of total carbonate in water samples was developed. The method is based on the color change of an acid-base indicator in relation to the concentration of permeable gas substances through a membrane. By using a new portable FIA system equipped with a gas-diffusion unit, a highly sensitive and on-site determination of total carbonate in aqueous solutions was investigated. A new color-change system with 4-(2',4'-dinitrophenylazo)-1-naphthol-5-sulfonic acid (DNN5S) was developed. Absorbance changes of the reagent solution were measured at 450 nm with a light-emitting diode (LED) as a light source. A new type of gas-diffusion unit was used, and was constructed with double tubing: the inner tubing was a micro porous PTFE (polytetrafluoroethylene) tubing (1.0 mm inner diameter and 1.8 mm outer diameter, pore size 2 microns, porosity 50%); the outer tubing was made of glass with 2.0 mm inner diameter. The optimized system conditions were as follows: the sample size was 200 microliters, the temperature of the air bath for the gas-diffusion unit was 25 degrees C, and the length of the gas-diffusion unit was 15 cm; each flow rate was 0.3 ml min-1. For measuring carbonate at low concentrations, a method for preparing water with less carbonate was proposed: the carbonate content of the water was decreased down to 5 x 10(-7) M. The calibration graph was rectilinear from 1 x 10(-6) M to 10(-3) M, and the detection limit (corresponding to a signal-to-noise ratio of 3) was 1 x 10(-6) M of carbonate. The relative standard deviation (RSD) of ten measurements of 2.3 x 10(-5) M Na2CO3 solution was 1.9%. The total carbonate in various kinds of water (such as river, sea, rain, distilled and ultra purified) was determined.

Carbonates↗

[Progress in targeting gene therapy].

In recent years the research on targeting in gene therapy has made much exciting progress. Many new technologies show great promising prospects such as the delivery of therapeutic gene to targeted cells by some antibodies or ligands, the use of specific gene promoter to tightly control gene expression within disease microenvironment, the use of some orally taken drugs for regulating expression time and expression level of DNA, and the targeted correction of deleted or mutated gene using RNA/DNA oligonucleotides. Yet, researchers still have a long way to go in order to reach the goal of applying the targeting gene therapy in clinical practice.

Gene Expression↗

[Telomere length and DCC gene mRNA expression of human large intestine cancers].

OBJECTIVE: To evaluate the role of telomere and DCC in tumor transformation and progression. METHODS: Telomere length and DCC gene mRNA expression were examined by southern blot hybridization and RT-PCR analysis in 46 adenomas of large intestine, 62 cancers of large intestine and corresponding normal mucosa. RESULTS: Shortening of the telomere was present in the tissues of 41.3% of the adenomas and 53.2% of the cancers, and their average TRF lengths were significantly shorter than those of corresponding normal mucosa(P<0.05, P<0.01), whereas the telomere elongation was only detected in 4.4% and 6.5% of the adenomas and cancers respectively. In addition, the average telomere length in colon carcinomas was also shorter than that in rectal carcinomas. Moreover, the average telomere lengths of the colorectal cancer mucosa became shorter with age. The rates of DCC mRNA expression deletion were 34.8% and 62.9% in the tissues of adenomas and cancers respectively. The DCC mRNA expression deletion occurred more frequently in poorly differentiated and Dukes C, D carcinomas than in well-differentiated and Dukes A, B carcinomas (P<0.05, P<0.01). However, no significant correlation was found between the length of telomere and the deletion of DCC mRNA expression in the cancers of large intestine. CONCLUSION: The telomere shortening and DCC mRNA deletion may represent the biologic behavior of transformation and development of the large intestine cancers.

Adult↗

[Effect of unprepared and prepared "he zi" on contraction of isolated trachea smooth muscle in rabbits].

Isolated trachea smooth muscle in rabbits induced by Ach or KCl was adopted to study the effect of unprepared and prepared "He Zi". The results showed that unprepared "He Zi" couldn't affect the contraction of isolated trachea smooth muscle in rabbit induced by Ach or KCl, and that prepared "He Zi" couldn't affect the contraction of isolated trachea smooth muscle in rabbit induced by KCl. The results also showed that prepared "He Zi" could significantly inhibit the contraction induced by Ach. The inhibition effect may not be associated with cGMP, adrenergic receptor, and release of NO and prostaglandin, and was epithelium independent. Maybe the different content of tannic acid contributed to the different effect between unprepared and prepared "He Zi".

Acetylcholine↗

[Tracing algorithms of ECG tracks from scanned image].

This paper presents a discussion on the tracing algorithms of the extraction of Electrocardiogram (ECG) data from scanned image of hardcopy ECG. We have advanced a new algorithm integrated by seek-and-predict method and polygon model method, which can tolerate crossing of two leads, fault-line and disturbing of grid lines. Two practical situations that may be encountered while processing hardcopy ECG are discussed in detail. Furthermore, the after-treatment of extraction-filtering is considered and an SVD-based filter designed to smooth ECG signal is analyzed.

Algorithms↗

Protease crosstalk with integrins: the urokinase receptor paradigm.

Migratory cells use both adhesion receptors and proteolytic enzymes to regulate their interaction with and response to extracellular matrices. Cooperation between integrins and proteases operates at several levels: integrin signaling induces proteases, proteases co-localize with integrins, and proteases regulate the interface between integrins and the intracellular cytoskeleton. One protease system intimately connected to integrins is the urokinase/urokinase receptor(uPAR)/plasmin system. Recent studies indicate urokinase promotes the ligand-like binding of its receptor to a set of beta1 and beta2 integrins, this binding in turn affecting integrin signaling and cell migration. The glycolipid anchor of uPAR associates with cholesterol-rich membrane rafts. Binding of uPAR to integrins may enrich integrin clusters with signaling molecules such as src-family kinases that localize to rafts and are important to integrin function. Signals derived from integrin/uPAR complexes promote the function of other integrins. Thus the urokinase/plasmin system coordinates with integrins to regulate cell: matrix interactions.

Endopeptidases↗

[The study of P68 RNA helicase on cell transformation].

The P68 nuclear protein is an ATP-dependent RNA helicase and RNA-dependent ATPase, and has been proved to have functions in the regulation of cell growth and division. Different phenotypes of P68 between tumouric cell lines (HeLa, TC3H10) and non-tumouric cell lines (NIH3T3 and NC3H10) were observed, and we reasoned that overexpression of P68 might profoundly affect cellular growth properties. We report here that overexpression human P68 by transfection of recombinant P68 expression vector in NIH3T3 and NC3H10 fibroblasts causes their tumorigenic transformation as determined by four criteria: (1) the morphology of transfectants from long fibric to spindle shaped; (2) formation of transformed foci on the monolayer of cells; (3) the transfectants acquired the ability to grow on soft agar forming cell microsphere cube; (4) tumor formation in nude mice after injection of transfected cells for 3-4 weeks.

3T3 Cells↗

[Anti-tumor immune response against mouse melanoma to xenogeneic vaccination].

OBJECTIVE: To study the inhibition of melanoma growth in mice by vaccination with xenogeneic melanocytes. METHODS: Xenogeneic vaccine was prepared from pig eye melanocytes. It was used before or after B16 melanoma challenge in C57 mice. The size of tumor was monitored. Cytotoxic T lymphocyte (CTL) activity of mouse spleen cells was measured by 51Cr release assay. Antibody response against pig melanocytes and B16 melanoma cells were detected by indirect ELISA. RESULTS: Preventive vaccination resulted in inhibition of tumor growth in 90% of the immunized mice, while therapeutic vaccination inhibited tumor growth in 50% of the treated mice. Specific CTL activity and antibodies in the immunized mice were detected. CONCLUSION: Anti-tumor immune response capable of inhibiting melanoma growth can be induced by xenogeneic melanocyte vaccination.

Animals↗

[Observation of preventing of bone loss during early postmenopause by percutaneous estradiol in Chinese postmenopausal women].

OBJECTIVE: To investigate the optimal regimen for application of percutaneous estradiol gel in preventing bone loss in Chinese postmenopausal women. METHODS: A 3-year open randomized clinical study was designed. The percutaneous estradiol gel was used in a cyclic regimen combined with micronized progesterone (MP) or medroxyprogesterone acetate (MPA). Sixty healthy women (naturally menopause for 1 to 5 years) were recruited and divided into four groups according to estrogen dosage and two kinds of progestin. All were given for 25 d/month. The cortical bone mineral density (BMD) of right radius was measured by single photon absorptiometry. The trabecular BMD in lumbar vertebrae was measured by quantitative CT. The spine and hip BMD were also measured by dual energy X-ray absorptiometry at baseline, 6, 12, 18, 24 and 36 months, respectively. The bone metabolic markers, scores of menopausal symptoms were also evaluated. RESULTS: Fifty-nine patients (98%) completed 1 year and 56 patients (93%) 2 years, 51 (85%) 3 years of study. The symptoms were alleviated by 80% after 6 months treatment on average. By the end of 24 month, the mean increases of BMD ranged from 4.3% to 7.5% in trabecular bone, and by the end of 36 month 4.2% to 6.2% in the lumbar (L) 2-4, 1.6% to 3.8% in the femur neck, with significant differences (P < 0.05). Comparing the 4 groups with each other, no significant differences (P > 0.05) were found in improvement of symptoms, bone markers and BMD. CONCLUSIONS: Both daily estrogen containing 0.75 mg and 1.5 mg E2 are effective to prevent the early postmenopausal bone loss and improve the menopausal symptoms. During 3 years treatment, the BMD of lumbar increased continuously and the BMD of hip increased in the first 2 years and then plateau.

Administration, Cutaneous↗

[Study of mechano-sensitive K+ channel in cerebral microvascular endothelial cells from rats].

This study was conducted to gain an understanding of the electrophysiologic characteristics of cerebral microvascular endothelial cells. The stress response of the potassium ion channel in cerebral microvascular endothelial cells was explored. We developed a model of the opening shear stress effect device and used it for calculation of shear stress. The cultured cerebral microvascular endothelial cells from rats were seeded on glass slide of 1 cm x 1 cm. K+ ion channel current was recorded by Patch-Clamp amplifier of Axonpatch 200A type and whole cell Patch-Clamp technique. The inward rectifier potassium current of cerebral microvessel endothelial cells has been elicited by flow stimulus of 1.1 dynes/cm2 shear stress, and there is good relationship between the current and voltage changing. The various stress responses of the cerebral vascular endothelial cells are related to the stress sensitive potassium ion channels of cellular membrane.

Animals↗