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Biomedical subjects

M Yan

Publications and source records attributed to M Yan.

At least 73 records · Page 4Linked to original sources

[Construction of porous hydroxyapatite (HA) block loaded with cultured chondrocytes].

OBJECTIVE: To construct a kind of bone healing enhancing implant with cultured chondrocytes bound to hydroxyapatite (HA). METHODS: Chondrocytes were obtained from the costicartilage of rat and were cultured on the porous HA blocks, 3 mm x 3 mm x 4 mm size, for three and seven days. Scanning electron micrograph was taken to show whether the cells grew outside and inside the pore of HA block. The cells cultured on tiny glass sheet for 2 days were used to prove where the cells come from by in situ hybridization technique with alpha1 (II) cDNA probe. RESULTS: Scanning electron micrographs showed that the pores of the HA surface and inside of the blocks are filled with cultured cells, especially the longer cultured block. The cells were chondrocytes confirmed by in situ hybridization. CONCLUSION: The porous HA can be used as cell cultured substrate and chondrocyte can adhere and proliferate inside the porous HA block.

Animals↗

Membrane-limited expression and regulation of Na+-H+ exchanger isoforms by P2 receptors in the rat submandibular gland duct.

1. Cell-specific reverse transcriptase-polymerase chain reaction (RT-PCR), immunolocalization and microspectrofluorometry were used to identify and localize the Na+-H+ exchanger (NHE) isoforms expressed in the submandibular gland (SMG) acinar and duct cells and their regulation by basolateral and luminal P2 receptors in the duct. 2. The molecular and immunofluorescence analysis showed that SMG acinar and duct cells expressed NHE1 in the basolateral membrane (BLM). Duct cells also expressed NHE2 and NHE3 in the luminal membrane (LM). 3. Expression of NHE3 was unequivocally established by the absence of staining in SMG from NHE3 knockout mice. NHE3 was expressed in the LM and in subluminal regions of the duct. 4. Measurement of the inhibition of NHE activity by the amiloride analogue HOE 694 (HOE) suggested expression of NHE1-like activity in the BLM and NHE2-like activity in the LM of the SMG duct. Several acute and chronic treatments tested failed to activate NHE activity with low affinity for HOE as expected for NHE3. Hence, the physiological function and role of NHE3 in the SMG duct is not clear at present. 5. Activation of P2 receptors resulted in activation of an NHE-independent, luminal H+ transport pathway that markedly and rapidly acidified the cells. This pathway could be blocked by luminal but not basolateral Ba2+. 6. Stimulation of P2U receptors expressed in the BLM activated largely NHE1-like activity, and stimulation of P2Z receptors expressed in the LM activated largely NHE2-like activity. 7. The interrelation between basolateral and luminal NHE activities and their respective regulation by P2U and P2Z receptors can be used to co-ordinate membrane transport events in the LM and BLM during active Na+ reabsorption by the SMG duct.

Adenosine Triphosphate↗

Vascular endothelial growth factor regulates endothelial cell survival through the phosphatidylinositol 3'-kinase/Akt signal transduction pathway. Requirement for Flk-1/KDR activation.

Vascular endothelial growth factor (VEGF) has been found to have various functions on endothelial cells, the most prominent of which is the induction of proliferation and differentiation. In this report we demonstrate that VEGF or a mutant, selectively binding to the Flk-1/KDR receptor, displayed high levels of survival activity, whereas Flt-1-specific ligands failed to promote survival of serum-starved primary human endothelial cells. This activity was blocked by the phosphatidylinositol 3'-kinase (PI3-kinase)-specific inhibitors wortmannin and LY294002. Endothelial cells cultured in the presence of VEGF and the Flk-1/KDR-selective VEGF mutant induced phosphorylation of the serine-threonine kinase Akt in a PI3-kinase-dependent manner. Akt activation was not detected in response to stimulation with placenta growth factor or an Flt-1-selective VEGF mutant. Furthermore, a constitutively active Akt was sufficient to promote survival of serum-starved endothelial cells in transient transfection experiments. In contrast, overexpression of a dominant-negative form of Akt blocked the survival effect of VEGF. These findings identify the Flk-1/KDR receptor and the PI3-kinase/Akt signal transduction pathway as crucial elements in the processes leading to endothelial cell survival induced by VEGF. Inhibition of apoptosis may represent a major aspect of the regulatory activity of VEGF on the vascular endothelium.

Amino Acid Chloromethyl Ketones↗

An MTP inhibitor that normalizes atherogenic lipoprotein levels in WHHL rabbits.

Patients with abetalipoproteinemia, a disease caused by defects in the microsomal triglyceride transfer protein (MTP), do not produce apolipoprotein B-containing lipoproteins. It was hypothesized that small molecule inhibitors of MTP would prevent the assembly and secretion of these atherogenic lipoproteins. To test this hypothesis, two compounds identified in a high-throughput screen for MTP inhibitors were used to direct the synthesis of a highly potent MTP inhibitor. This molecule (compound 9) inhibited the production of lipoprotein particles in rodent models and normalized plasma lipoprotein levels in Watanabe-heritable hyperlipidemic (WHHL) rabbits, which are a model for human homozygous familial hypercholesterolemia. These results suggest that compound 9, or derivatives thereof, has potential applications for the therapeutic lowering of atherogenic lipoprotein levels in humans.

Alanine Transaminase↗

A novel approach to distinguish between intron-containing and intronless genes based on the format of Z curves.

A novel method to distinguish between intron-containing and intronless DNA sequences has been proposed, based on different statistic behaviors between them. In this method, DNA sequences are first represented as Z curves. Three exponents alpha, beta and gamma for each given sequence are calculated based on the format of the Z curve for the DNA sequence. A three-dimensional space is spanned by the three exponents. Each DNA sequence may be represented by a point in this space. One hundred intronless and intron-containing genes, respectively, were selected randomly from the GenBank or EMBL database. It is shown that the 200 points are roughly distributed in different regions. The best separating plane to separate the two regions is obtained by using Fisher's discriminant algorithm. For any given sequence to be discriminated, calculate three exponents alpha, beta and gamma, corresponding to a point in the three-dimensional space. If the point is situated at the upper region of the separating plane, the sequence is discriminated as an intronless one; otherwise, the sequence is an intron-containing one. A test of the method for the sequences in an independent test set shows that the discriminant accuracy reaches as high as 89.0%.

Animals↗

Regional gray matter, white matter, and cerebrospinal fluid distributions in schizophrenic patients, their siblings, and controls.

BACKGROUND: Cortical gray matter volume reductions and cerebrospinal fluid (CSF) volume increases are robust correlates of schizophrenia, but their sources have not been established conclusively. METHODS: Structured diagnostic interviews and magnetic resonance imaging scans of the brain were obtained on 75 psychotic probands (63 with schizophrenia and 12 with schizoaffective disorder), ascertained so as to be representative of all such probands in a Helsinki, Finland, birth cohort; 60 of their nonpsychotic full siblings; and 56 demographically similar control subjects without a personal or family history of treated psychiatric morbidity. RESULTS: Patients with schizophrenia and their siblings exhibited significant reductions in cortical gray matter volume and significant increases in sulcal CSF volume compared with controls. The patients, but not their siblings, also exhibited significant reductions in white matter volume and significant increases in ventricular CSF volume. Regional effects were most robust when component volumes were expressed as percentages of overall regional volumes; in this case, for patient and sibling groups, gray matter volume reductions and sulcal CSF volume increases were significantly more pronounced in the frontal and temporal lobes than in the remainder of the brain. None of the group differences varied significantly by sex or hemisphere. CONCLUSIONS: Structural alterations of the cerebral cortex, particularly in the frontal and temporal lobes, are present in patients with schizophrenia and in some of their siblings without schizophrenia; such changes are thus likely to reflect genetic (or shared environmental) effects. Ventricular enlargement is unique to the clinical phenotype and is thus likely to be affected primarily by nonshared causative factors.

Adult↗

Jak-STAT pathway is involved in the induction of TNF-beta gene during stimulation by IL-2.

IL-2 is the major regulatory cytokine of the immune system. It plays a key role in T cell survival, growth and activation. IL-2 may induce the expression of multiple genes including some cytokine genes. The induction of these genes is triggered by different signal pathways, one of them being the Jak-STAT signal pathway. The genes regulated by this pathway remain to be determined. By studying IL-2-inducible genes, we have confirmed that the TNF-beta gene is one of the immediate early genes activated by IL-2. By analysis of the DNA sequences around 180-300 bases upstream of the transcription initiation point of the mouse TNF-beta gene, we demonstrate that there is a STAT5 binding site which is essential to the inducibility of the TNF-beta gene. Furthermore, in BA/F3 cells co-transfected with the STAT5A gene and IL-2R beta gene, the activation of the TNF-beta gene promoter by IL-2 was greatly promoted, whereas the TNF-beta gene promoter became IL-2-non-inducible if the STAT5A gene was substituted with a dominant negative STAT5A, i.e. a C-terminally truncated mutant. Taken together, our results show that the Jak-STAT signal pathway is involved in induction of the TNF-beta gene in cells stimulated by IL-2.

Animals↗

Effects of micromolar concentrations of Mn, Mo, and Si on alpha1-adrenoceptor-mediated contraction in porcine coronary artery.

We studied the effects of trace elements, Mn, Mo, and Si, on vasoconstriction induced by norepinephrine (NE) or electrical field stimulation in isolated porcine right coronary arteries. Alpha1-adrenoceptor (AR) antagonist prazosin dose-despondently suppressed vasoconstriction in response to NE or field stimulation indicating an alpha1-AR mediated response. Mn, Mo, and Si at 0.3-3 micromol/L dose-despondently inhibited NE mediated contraction (all p < 0.05). In contrast, Mn, Mo, and Si at the same concentrations (0.3-3 micromol/L) enhanced the maximal contractile response to field stimulation in a dose-dependent manner (all p < 0.05), but these elements at 10 micromol/L suppressed the vasoconstrictive response. The results indicate that in porcine right coronary arteries, the alpha1-AR-mediated vasoconstriction by NE or electrical field stimulation was affected differently by micromolar concentrations of Mn, Mo, and Si and that the elements might facilitate NE release presynaptically but inhibit the contractile response postsynaptically.

Adrenergic alpha-Antagonists↗

A new fourier transform approach for protein coding measure based on the format of the Z curve.

MOTIVATION: At the core of most protein gene-finding algorithms are the coding measures used to make a decision on coding/non-coding. Of the protein coding measures, the Fourier measure is one of the most important. However, due to the limited length of the windows usually used, the accuracy of the measure is not satisfactory. This paper is devoted to improving the accuracy by lengthening the sequence to amplify the periodicity of 3 in the coding regions. RESULTS: A new algorithm is presented called the lengthen-shuffle Fourier transform algorithm. For the same window length, the percentage accuracy of the new algorithm is 6-7% higher than that of the ordinary Fourier transform algorithm. The resulting percentage accuracy (average of specificity and sensitivity) of the new measure is 84.9% for the window length 162 bp. AVAILABILITY: The program is available on request fromC.-T. Zhang. CONTACT: ctzhang@tju.edu.cn

Algorithms↗

Prediction of the helix/strand content of globular proteins based on their primary sequences.

An improved multiple linear regression method has been proposed to predict the content of alpha-helix and beta-strand of a globular protein based on its primary sequence. The amino acid composition and the auto-correlation functions based on the hydrophobicity profile of the primary sequence have been taken into account in the algorithm. The resubstitution test shows that the average absolute errors are 0.077 and 0.073 with the standard deviations 0.059 and 0.057 for the prediction of the content of alpha-helix and beta-strand, respectively. A stringent cross-validation test, i.e., the jackknife test, shows that the average absolute errors are 0.087 and 0.081 with the standard deviations 0.067 and 0.065 for the prediction of the content of alpha-helix and beta-strand, respectively. Both tests indicate the self-consistency and the extrapolating effectiveness of the new algorithm. This greatly improves on previous results (Eisenhaber,F., Imperiale,F., Argos,P. and Frommel,C., 1996, Proteins, 25, 157-168). Compared with other methods currently available, our method has the merits of simplicity and ease-of-use as well as a higher prediction accuracy. The only input of the method is the primary sequence of the query protein to be predicted. The program is available on request via e-mail: ctzhang@tju.edu.cn.

Amino Acid Sequence↗

Functional assessment of alpha 1-adrenoceptor subtypes in porcine coronary artery.

1. alpha 1-Adrenoceptors are known to play an important role in vasoconstriction in response to adrenergic stimulation. However, the functional importance of alpha 1-adrenoceptor subtypes at the epicardial coronary artery remains unclear. We examined alpha 1-adrenoceptor subtypes by comparing functional affinities for alpha-adrenoceptor antagonists on noradrenaline (NA)-induced vasoconstriction in porcine denuded right coronary arteries. 2. Noradrenaline induced a dose-dependent vasoconstriction in incubated vessel rings. Prazosin and phentolamine were potent and competitive antagonists for NA-induced contraction (pA2 10.27 and 9.03, respectively). In contrast, the selective alpha 2-adrenoceptor antagonist yohimbine had a low affinity (pA2 6.13). Two selective alpha 1A-adrenoceptor antagonists, WB 4101 and 5-methyl urapidil, were potent and competitive antagonists of alpha 1-adrenoceptor-induced contraction (pA2 10.67 and 8.90, respectively) and the selective alpha 1D-adrenoceptor antagonist BMY 7378 had a low affinity (pA2 6.06). Noradrenaline-induced contraction was insensitive to the alkylating effects of chlorethylclonidine. These observations indicate that the vasoconstriction is predominantly mediated by the alpha 1A-adrenoceptor subtype. This was also supported by a good correlation between pA2 values from the present study and reported binding affinities (pKi) of various alpha-adrenoceptor antagonists with cloned human alpha 1A-adrenoceptors (r = 0.98), but not for alpha 1B- or alpha 1D-adrenoceptor subtypes (r = 0.77 and 0.41, respectively). 3. Our results indicate that the alpha 1A-adrenoceptor is the main functional receptor subtype in porcine denuded coronary arteries.

Adrenergic alpha-Agonists↗

Gypsum-bonded alumina dental investment for high-fusing casting.

In this study, we developed a new gypsum-bonded investment for high-fusing alloys. The investment was composed of gypsum as a binder and alumina as a refractory. Effects of type of alumina powder and gypsum content on characteristics of the gypsum-bonded alumina investment were investigated. Obtained characteristics of this experimental investment were as follows: fluidities ranged from 48.8 to 88.9 mm; setting times ranged from 21.2 to more than 120 minutes; setting expansions ranged from 0.4 to 1.3%; green strengths showed 0.5 to 4.5 MPa; fired strengths ranged from 0.2 to 1.7 MPa; thermal expansions after firing were -1.60 to 2.16%. Thermal expansion occurred because of the chemical reaction between Al2O3 and CaO decomposed from gypsum. These results suggest that this gypsum-bonded alumina investment with 20 or 25 mass% gypsum content possessed the fundamental properties for high-fusing alloy casting.

Aluminum Oxide↗

Effects of magnesia and potassium sulfate on gypsum-bonded alumina dental investment for high-fusing casting.

The purpose of this study was to improve the characteristics of gypsum-bonded alumina investments using magnesia and potassium sulfate as chemical additives. Magnesia content improved fluidity, delayed setting reaction, increased green strength, and decreased setting expansion, when mixed with distilled water. When the investment was mixed with potassium sulfate, the setting time and setting expansion were reduced, and the thermal expansion increased, however, the green strength decreased. Therefore, the investment with a small amount of magnesia mixed with potassium sulfate was considered a suitable composition, having adequate setting behavior, enough green strength and sufficient compensate expansion for casting.

Aluminum Oxide↗

[The role of lens epithelium in cataract formation in diabetic rats].

This study was designed to evaluate the changes and the role of lens epithelium in sugar cataract formation, in regard to the fact that the highest level of aldose reductase is found in this layer of lens. By light and electron microscopy, we examined the histological changes of central epithelium in lens of rats made diabetic with streptozotocin (STZ) with or without AL1576, an aldose reductase inhibitor, at varying periods of time ranging from 5 to 40 days after intraperitoneal injection of STZ. Also, we examined Na-K-ATPase activity in lens epithelium of rats with diabetes, diabetes plus AL1576 and normal controls at the time of 30 days. The results showed that the first detectable abnormalities occurred after 15 days of STZ injection and were limited to the lens epithelium; cell edema, intracellular vacuoles and extention of rough endoplasmic reticulum pool were remarkable; that AL1576 could prevent almost all of the lesion mentioned above; and that Na-K-ATPase activity in lens epithelium of rats with diabetes increased at the time of 30 days. The findings suggest that lens epithelium may play an important role in sugar cataractogenesis.

Aldehyde Reductase↗

[Observation on inhibitory effect of Coptis alone and its combination with Scutellaria and Liquorice on the growth of Staphylococcus aureus].

Using MIC as relative index, an observation was made on the growth inhibition of Staphylococcus aureus by Coptis alone or in combination with Scutellaria and or liquorice. The result shows that when Coptis is combined with Scutellaria, the bacteriostatic effect gets weaker for Scutellaria, but remains unchanged for Coptis, and when Scutellaria takes greater percentages in the combination, the effect of Coptis seems to be stronger. When Coptis is combined with liquorice, whatever the respective percentages may be, the effect tends to get weaker. When the three drugs are used in combination, the effect remains unchanged or even gets stronger for Coptis or liquorice, but tends to decrease for Scutellaria. Coptis used singly or in combination with Scutellaria or liquorice is inferior in bacteriostatic effect to the combination of the three drugs. The mechanism is yet to be further studied.

Drug Combinations↗

[Influence of alum on intestinal flora in mice].

OBJECTIVE: To observe the influence of alum on the intestinal microecological balance in normal microorganisms. METHOD: The mice were administered orally with alum of a small dosage(0.25/kg) and a large dosage(1 g/kg) for half a month, two months and three months, and a micro flora analysis of the mice was carried out at intervals of the above mentioned administrations. RESULT: The intestinal flora in the animals administered with alum was imbalanced. The counts of bifidobacteria and lactobacilli closely related to human physiological activities were decreased. The counts of pathogenic E. Coli significantly increased; and the longer the animals were treated with alum, the stronger the microecological balance was influenced. CONCLUSION: Alum could induce imbalance of the normal intestinal flora in mice.

Alum Compounds↗

Evidence that microsomal triglyceride transfer protein is limiting in the production of apolipoprotein B-containing lipoproteins in hepatic cells.

The microsomal triglyceride transfer protein (MTP) is a heterodimeric lipid transfer protein that is required for the assembly and secretion of apolipoprotein B (apoB)-containing lipoproteins. A key unresolved question is whether the MTP-mediated step is rate limiting. To address this, a unique experimental strategy was used that allowed the in situ modulation and measurement of MTP triglyceride transfer activity. In order to accomplish this, an irreversible photoaffinity inhibitor, BMS-192951, was designed and synthesized. When incubated with purified MTP and irradiated with UV light at 360 nm, BMS-192951 inhibits triglyceride transfer by covalently binding to the protein. HepG2 cells were treated with either increasing concentrations of BMS-192951 (0-15 microM) with 5 min of ultraviolet irradiation, or 3.0 microM BMS-192951 with various lengths (0-15 min) of ultraviolet irradiation. Microsomal extracts were prepared exhaustively dialyzed to remove unbound inhibitor, and assayed for MTP-mediated triglyceride transfer activity. BMS-192951 was shown to reduce MTP activity in both a dose- and UV exposure time-dependent fashion. Measurement of apoB concentration in the media showed that apoB secretion was reduced in proportion to the in situ inhibition of MTP activity, while no change was observed in apoA-I secretion. Experiments performed in McArdle RH-7777 rat hepatoma cells and primary rat hepatocytes gave nearly identical results; the decrease in apoB secretion was proportional to the decrease in MTP activity. These results indicate that MTP-mediated lipid transfer is limiting in the assembly and secretion of apoB-containing lipoproteins in hepatic cells under the conditions tested.

Affinity Labels↗

Multiple ATP-dependent steps in RNA polymerase II promoter melting and initiation.

Permanganate probing and abortive initiation assays were used to investigate the role of ATP in several successive stages of transcription initiation at the activated adeno E4 and mouse DHFR promoters. Removal of ATP at several points along the multi-step pathway blocked further progress towards its completion. Most strikingly, even if the DNA transcription start site is opened using ATP, the subsequent removal of ATP disallows formation of the first phosphodiester bond of the RNA. After ATP-dependent formation of a short RNA, a new transcription complex forms, which is more stable and has a longer open region. Both RNA and ATP appear to play roles in the formation of this complex. The need for ATP throughout this multi-step initiation pathway leads to new and unexpected possibilities for the use of energy and ATPases in transcription initiation.

Adenosine Triphosphatases↗