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Biomedical subjects

L Lei

Publications and source records attributed to L Lei.

At least 37 records · Page 2Linked to original sources

[Separation of echinacoside by reversed-phase preparative high performance liquid chromatography].

Echinacoside, one kind of phenylethanoid glycosides (PhGs), was isolated from the stems of Cistanche tubulosa (Schenk) R. Wight with a series of steps, including solvent extraction, D101 polymer adsorption column separation, Sephadex LH-20 separation, C18 column reversed-phase preparative high performance liquid chromatographic (RP-prep-HPLC) preparation and polyamide thin-layer chromatographic detection. The purity of the product was over 98%. The chemical structure of echinacoside was identified by 1H NMR and 13C NMR. To find out the optimum condition of mobile phase of RP-prep-HPLC, several systems were used in this work. Finally acetonitrile-1% HCOOH water (18:82, V/V) system was found to be the best. On the other hand D101 polymer adsorption column and Sephadex LH-20 were also effective for PhGs separation.

Chromatography, High Pressure Liquid↗

[Metabolic regulation of phenylethanoid glycosides from Herba cistanches in dogs' gastrointestine].

AIM: To study the metabolic process of phenylethanoid glycosides (PhGs) in the gastrointestine of beagle dogs that were administered intragastrially process, and develop some new methods of biopharmacology on the effective position of traditional Chinese medicine. METHODS: High-performance liquid chromatography was used to purify constituents from faeces and analyze relative contents of the three main compounds in the gastrointestinal tract at different times and in the faeces of dogs. Every sample was collected, extracted with methanol and analyzed with integration. RESULTS: Four compounds, based on reference substances, were identified as echinacoside, acteoside, isoacteoside, and 2'-acetylacteoside from extraction of faces of dogs. Quantitative "with HPLC" analysis reveals that variation of ratios of the three main compounds is not distinct when moving in the gastrointestinal tract 7 h, that is quite different from those in faeces, in which the content of echinacoside fell from 48.0% to 16.0%, and acteoside increased from 11.0% to 34.7%. CONCLUSION: PhGs are mainly metabolized in large intestine. Among them, a portion of echinacoside is transformed into aceteoside.

Animals↗

[Biochemical remodeling of myocardial collagen and its relation to cardiac renin-angiotensin system in rats after myocardial infarction].

OBJECTIVE: This study aimed at the biochemical remodeling of myocardial collagen and its relation to cardiac renin-angiotensin system (RAS) after myocardial infarction(MI). METHODS: The ventricular and plasma angiotensin II (Ang II) content, total collagen content, and collagen concentration in left ventricle(LV) and right ventricle(RV) of MI and sham-operated control(SOC) rats on 3, 15 and 42 days after operation were measured dynamically by means of radioimmunoassay etc. RESULTS: The plasma Ang II content increased only in MI rats on day 3. The Ang II content in LV of the MI group increased markedly on day 3 after MI as compared with that of the SOC group. It sustained at this higher level on the 15th and 42th days, and the similar changes were observed in RV as well. The biochemical remodeling of collagen in LV and RV occurred after MI and the remodeling of collagen showed similar patterns with its Ang II content, except that collagen remodeling in RV was delayed than the change of Ang II content. CONCLUSION: The cardiac RAS was obviously activated after MI. Ventricular Ang II might play an important role in the biochemical remodeling of myocardial collagen, and its role was independent of circulating RAS.

Angiotensin II↗

[Beta-actin changes during hair cell regeneration in the chick basilar papilla].

OBJECTIVE: To investigate the changes of beta-actin and beta-actin mRNA expression during hair cell regeneration in the chick basilar papilla(BP) following gentamicin ototoxicity. METHODS: Thirty-six chicks were given muscular injection of gentamicin at dosage of 100 mg/kg.d for 10 consecutive days, and twelve additional chicks received injection of normal saline instead of gentamicin as controls. At 1, 3, 7, 14, 21 and 28 days after the treatment, six chicks in experimental group and two chicks in control group were sacrificed. Basilar papillas were prepared for immunohistochemistry and in situ hybridization study. beta-actin and beta-actin mRNA expression in the damaged region were histologically analyzed by computer image system. RESULTS: beta-actin immunoreactivity of BP changed significantly after treatment with gentamicin. From the first day to the fourteenth day after gentamincin, beta-actin immunoreactivity turned positive gradually. It approximated the normal response level at the fourteenth day after treatment of gentamincin. Then increasing rate of beta-actin immunoreactivity positive response level slowed from the fourteenth day after the treatment. At seventh day after treatment, beta-actin mRNA expression in BP increased to peak value, which preceded the peak expression of beta-actin in the regenerated hair cells. CONCLUSION: Nascent hair cells seem to mature 14 days after hair cell damage, which may play a role in auditory functional recovery.

Actins↗

[Effect of p14ARF gene on cell growth of human laryngeal tumor cells and expression of endogenous p53 protein].

OBJECTIVE: To explore the inhibitory effect of p14ARF on the cell growth of laryngeal carcinoma and the expression on endogenous p53. METHODS: p14ARF cDNA was transferred to the cell line Hep-2 of squamous cell carcinoma of the larynx by gene transfer to study the cell cycles and the expression of endogenous wild type p53 using flow cytometry, polymerase chain reaction(PCR) and Western-Blotting. RESULTS: Expression of p14ARF significantly affected the Hep-2 cell growth. The clone-forming efficiency of the Hep-2 cells transferred with p14ARF was 57%, compared with empty vector pcDNA3. The number of 48 hours after transfer with p14ARF cDNA at both G0/G1 and G2/M was two fold as the control. The expression of endogenous wild type p53 was significantly enhanced. CONCLUSION: Expression of p14ARF can up-regulate the expression of endogenous wild type p53 and inhibit the Hep-2 cell growth of human laryngeal squamous cell carcinoma at both G0/G1 and G2/M.

Carcinoma, Squamous Cell↗

Yohimbine dimers exhibiting binding selectivities for human alpha2a- versus alpha2b-adrenergic receptors.

A series of yohimbine dimers was prepared and evaluated at the human alpha2a- and alpha2b-adrenergic receptors (ARs) expressed in Chinese hamster ovary (CHO) cells. All dimers display higher binding selectivities for alpha2a versus alpha2b subtype than yohimbine, and four compounds (3d, 3e, 3g and 3i) represent the most potent and alpha2a versus alpha2b-AR selective ligands identified so far.

Adrenergic alpha-2 Receptor Antagonists↗

Dietary intake of isothiocyanates: evidence of a joint effect with glutathione S-transferase polymorphisms in lung cancer risk.

Isothiocyanates (ITCs) are nonnutrient compounds in cruciferous vegetables with anticarcinogenic properties. One proposed mechanism for their protective action is through down-regulation of cytochrome P-450 biotransformation enzyme levels and induction of phase II detoxifying enzymes. Because ITCs also serve as a substrate for GSTs, we evaluated dietary intake of ITCs and GSTM1 and GSTT1 genotype information in a lung cancer case-control study. There were 503 newly diagnosed lung cancer cases (264 men and 239 women) identified from The University of Texas M. D. Anderson Cancer Center and 465 controls (252 men and 213 women) recruited from enrollees in a local managed care organization. Subjects had an in-person interview including a semiquantitative food frequency questionnaire, and blood samples were obtained for genotyping. Cases reported significantly lower ITC intake per day compared with controls (P = 0.009). There was no main effect associated with the GSTM1 null genotype [odds ratio (OR) = 1.09]. However, there was a statistically significant OR of 1.41 associated with the GSTT1 null genotype. On stratified analysis, low ITC intake and the GSTM1 null genotype were associated with increased lung cancer risk in current smokers, with an OR of 2.22 [confidence interval (CI) = 1.20-4.10). For current smokers with the GSTT1 null genotype, the OR with low ITC intake was 3.19 (CI = 1.54-6.62). The comparable OR in the presence of both null genotypes was 5.45 (CI = 1.72-17.22). These effects were not demonstrable for former smokers by GSTM1 genotype, although the risk for low ITC intake and GSTT1 null genotype was 1.79 (CI = 0.95-3.37). Thus, current smokers who are homozygous null for the GST null genotype and who consume less carcinogenic blocking compounds are at higher lung cancer risk. Some of the inconsistencies reported in the role of GST genotypes in lung cancer risk could be due to unexpected confounding from dietary factors.

Aged↗

Construction and analytical application of ion-selective piezoelectric sensor for atropine sulfate.

The method describes the use of a piezoelectric quartz crystal (PQC) as a substitute for ion-selective electrodes. The approach is feasible when the membrane materials are electrically non-conductive and membrane potential measurements are consequently not possible. An ion-selective piezoelectric sensor sensitive to atropine sulfate was constructed by coating a PVC membrane containing activant on one the side of a PQC. On the basis of selective adsorption of atropine ions across the modified film and the sensitive mass response of PQC, the method exhibits a sensitive, rapid response and is easy to operate without pretreatment of the sample. The logarithm of the frequency shift gave a linear relationship with the logarithm of atropine sulfate concentration in the 1.0 x 10(-8)-1.0 x 10(-3) M range with a detection limit of 5.0 x 10(-9) M at pH 7.0. Recoveries were from 98.7-102.2%. Two activants, atropine tetraphenylborate and atropine dipicrylaminate, were synthesized and investigated. Influencing factors were also examined and optimized. The results for real samples obtained by the proposed method agreed with those obtained by conventional methods.

Atropine↗

Endomorphin-1 reduces carrageenan-induced fos expression in the rat spinal dorsal horn.

Intraplantar injection of carrageenan induced significant Fos expression in the superficial and deep spinal dorsal horn at the L(4)-L(5)segments and extensive peripheral edema of the ipsilateral foot in rats. Intraplantar injection of endomorphin-1, endogenous ligand for mu opioid receptor, in the same region produced dose-dependent reduction of carrageenan-induced Fos expression and peripheral edema, which were completely blocked by co-administration of intraplantar injection of naloxone (20 microgram). The systemic injection of the highest dose of endomorphin-1 (50 microgram) had no significant reductory effect on Fos expression and peripheral edema. These results further provided a strong evidence for involvement of mu opioid receptor in peripheral analgesia, particularly in inflammation pain.

Analgesics, Opioid↗

A region within the RAP74 subunit of human transcription factor IIF is critical for initiation but dispensable for complex assembly.

Human transcription factor IIF (TFIIF) is an alpha(2)beta(2) heterotetramer of RNA polymerase II-associating 74 (RAP74) and RAP30 subunits. Mutagenic analysis shows that the N-terminal region of RAP74 between L155 (leucine at codon 155) and M177 is important for initiation. Mutants in this region have reduced activity in transcription, but none are inactive. Single amino acid substitutions at hydrophobic residues L155, W164, I176, and M177 have similar activity to RAP74(1-158), from which all but three amino acids of this region are deleted. Residual activity can be explained because each of these mutants forms a complex with RAP30 and recruits RNA polymerase II into the preinitiation complex. Mutants are defective for formation of the first phosphodiester bond from the adenovirus major late promoter but do not appear to have an additional significant defect in promoter escape. Negative DNA supercoiling partially compensates for the defects of TFIIF mutants in initiation, indicating that TFIIF may help to untwist the DNA helix for initiation.

Adenoviridae↗

The RAP74 subunit of human transcription factor IIF has similar roles in initiation and elongation.

Transcription factor IIF (TFIIF) is a protein allosteric effector for RNA polymerase II during the initiation and elongation phases of the transcription cycle. In initiation, TFIIF induces promoter DNA to wrap almost a full turn around RNA polymerase II in a complex that includes the general transcription factors TATA-binding protein, TFIIB, and TFIIE. During elongation, TFIIF also supports a more active conformation of RNA polymerase II. This conformational model for elongation is supported by three lines of experimental evidence. First, a region within the RNA polymerase II-associating protein 74 (RAP74) subunit of TFIIF (amino acids T154 to M177), a region that is critical for isomerization of the preinitiation complex, is also critical for elongation stimulation. Amino acid substitutions within this region are shown to have very similar effects on initiation and elongation, and mutagenic analysis indicates that L155, W164, N172, I176, and M177 are the most important residues in this region for transcription. Second, TFIIF is shown to have a higher affinity for rapidly elongating RNA polymerase II than for the stalled elongation complex, indicating that RNA polymerase II alternates between active and inactive states during elongation and that TFIIF stimulates elongation by supporting the active conformational state of RNA polymerase II. The deleterious I176A substitution in the critical region of RAP74 decreases the affinity of TFIIF for the active form of the elongation complex. Third, TFIIF is shown by Arrhenius analysis to stimulate elongation by populating an activated state of RNA polymerase II.

Amino Acid Sequence↗

[Study of bacterial adhesion to prosthetic valve materials in vitro].

This article reports a study of the bacterial adhesion to prosthetic valve materials in vitro. The method for assessing the degree of bacterial adhesion to prosthetic valve materials was established primarily. The capacities of staphylococcus aureus(SA), staphylococcus epidermidis(SE), Escherichia Coli(EC) and Pseudomonas aeruginosa(PA) for adhesion to Dacron, Pyrolytic carbon and polytetrofluoroethylene (PTFE) were quantitatively determined by the plate counting and Gamma-ray counting of 125I radiolabeled bacteria in vitro. The results showed that the capacities of four types of bacteria for adhesion to Dacron, Pyrolytic carbon and PTFE coincided with the bacterial-growth curve. The capacities of four bacteria for adhering to Dacron were stronger. The adhesion of SE to Pyrolytic carbon was the strongest. The adhesion of PA Kept up a high level. The capacities of EC and PA for adhering to PTFE were the strongest. The results indicate that the capacities of different types of bacteria for adhesion to the same prosthetic valve material are different, and the capacities of one type of bacteria for adhesion to different materials are also different.

Bacterial Adhesion↗

[Determination of electron density in Ar-air mixed coolant gas ICP].

The lateral distribution of electron number density at 0, 2, 5, 10 mm above the load coil in Ar-air mixed coolant gas ICP was measured with Stark broadening method of Hbeta line (486.1 nm) and some of the lateral distribution was converted into radial distribution by Abel inversion under the condition similar to that in the low power Ar-ICP. The results show that the distribution of electron number in Ar-air mixed coolant gas ICP is similar to that in Ar-ICP and exhibit off axis maxima. When a lot of air was added to the coolant, the positions of the off axis maxima would near the axis. The electron number density reached maximum and was higher than that in Air-ICP as 5%-10% air was added to the coolant.

English Abstract↗

Sonic hedgehog is essential to foregut development.

Congenital malformation of the foregut is common in humans, with an estimated incidence of 1 in 3000 live births, although its aetiology remains largely unknown. Mice with a targeted deletion of Sonic hedgehog (Shh) have foregut defects that are apparent as early as embryonic day 9.5, when the tracheal diverticulum begins to outgrow. Homozygous Shh-null mutant mice show oesophageal atresia/stenosis, tracheo-oesophageal fistula and tracheal and lung anomalies, features similar to those observed in humans with foregut defects. The lung mesenchyme shows enhanced cell death, decreased cell proliferation and downregulation of Shh target genes. These results indicate that Shh is required for the growth and differentiation of the oesophagus, trachea and lung, and suggest that mutations in SHH and its signalling components may be involved in foregut defects in humans.

Animals↗

Activation of L-type calcium channel by tolazoline derivatives: role of isothiocyanate moiety.

Studies have investigated the pharmacologic mechanism of 2-(4'-isothiocyanatobenzyl) imidazoline (IBI) and analogs for interaction with imidazoline receptors (IRs), alpha-adrenergic receptors (alpha-ARs), and calcium channels in cardiovascular muscle systems. IBI differs from tolazoline by substitution of an electrophilic isothiocyanato (NCS) group. Unlike tolazoline, which is a partial alpha-AR agonist, IBI produced an irreversible, slow-onset, and sustained contraction of rat aorta with an median effective concentration (EC50) value of 5 microM, and a maximal contraction (116%) greater than that of phenylephrine (100%) and tolazoline (59%). The IBI-induced contractions were dependent on calcium channels and independent of alpha-ARs or IRs. Similarly, structure-activity relation studies in rat aortic smooth muscles on a series of synthesized IBI analogs indicated that NCS analogs, but not those without the NCS group, exhibited effects by a non-alpha-AR, non-IR, but a calcium channel-dependent mechanism. Thus the presence of an intact IBI ring in these analogs is not a requirement for these activities. Further, IBI inhibited dihydropyridine (DHP, [3H]PN 200-110 and [3H]Bay K 8644) binding to L-type calcium channels of T-tubule membranes in rabbit skeletal muscle. In contrast to nifedipine, IBI and NCS derivatives (nifedipine-NCS, naphazoline-NCS) only partially (50-88%) displaced specific binding of these radioligands. A single site of noncooperative interaction was observed for nifedipine (nH = 0.97), whereas tolazoline-NCS (IBI, nH = 1.46) and nifedipine-NCS (nH = 1.37) exhibited a positive cooperativity in binding to DHP sites. These receptor-binding data indicate that NCS derivatives bind to L-type calcium channels and interact allosterically with DHP-binding sites. Direct binding of the NCS group to specific nucleophilic protein sites of the calcium channel may be responsible for its activation and the subsequent contractile effects of IBI.

Adrenergic alpha-Agonists↗

Functions of the N- and C-terminal domains of human RAP74 in transcriptional initiation, elongation, and recycling of RNA polymerase II.

Transcription factor IIF (TFIIF) cooperates with RNA polymerase II (pol II) during multiple stages of the transcription cycle including preinitiation complex assembly, initiation, elongation, and possibly termination and recycling. Human TFIIF appears to be an alpha2beta2 heterotetramer of RNA polymerase II-associating protein 74- and 30-kDa subunits (RAP74 and RAP30). From inspection of its 517-amino-acid (aa) sequence, the RAP74 subunit appears to comprise separate N- and C-terminal domains connected by a flexible loop. In this study, we present functional data that strongly support this model for RAP74 architecture and further show that the N- and C-terminal domains and the central loop of RAP74 have distinct roles during separate phases of the transcription cycle. The N-terminal domain of RAP74 (minimally aa 1 to 172) is sufficient to deliver pol II into a complex formed on the adenovirus major late promoter with the TATA-binding protein, TFIIB, and RAP30. A more complete N-terminal domain fragment (aa 1 to 217) strongly stimulates both accurate initiation and elongation by pol II. The region of RAP74 between aa 172 and 205 and a subregion between aa 170 and 178 are critical for both accurate initiation and elongation, and mutations in these regions have similar effects on initiation and elongation. Based on these observations, RAP74 appears to have similar functions in initiation and elongation. The central region and the C-terminal domain of RAP74 do not contribute strongly to single-round accurate initiation or elongation stimulation but do stimulate multiple-round transcription in an extract system.

Amino Acid Sequence↗

[Treatment of metastatic bone pain with Aredia(Pamidronate)].

OBJECTIVE: To evaluate the efficacy of Aredia to relieve pain due to metastatic bone tumor. METHODS: Aredia 60-90 mg in 500 ml 5% glucose or normal saline was given by i.v. infusion. The treatment was repeated at 2-4 week intervals. RESULTS: Complete relief of pain was observed in 9 patients, partial relief in 13 patients. The treatment was of no effect in 3 patients. The therapeutic effect was achieved in 72.7% of the patients within 2 weeks. CONCLUSION: Aredia is a good pain reliever in patients with metastatic cancer of the bone.

Adult↗

Medetomidine analogs as alpha 2-adrenergic ligands. 3. Synthesis and biological evaluation of a new series of medetomidine analogs and their potential binding interactions with alpha 2-adrenoceptors involving a "methyl pocket".

The synthesis and the biological evaluation of a new series of medetomidine analogs are reported. The substitution pattern at the phenyl ring of the tetralin analogs had a distinct influence on the alpha 2-adrenoceptor binding affinity. 4-Methylindan analog 6 was the most potent alpha 2-adrenoceptor binding ligand among these 4-substituted imidazoles, and its alpha 2-adrenoceptor selectivity was greater than the 5-methyl tetralin analog 4c. Ligand-pharmacophore and receptor modeling were combined to rationalize alpha 2-adrenoceptor binding data of the imidazole analogs in terms of ligand-receptor interactions. The structure-activity relationships that were apparent from this and previous studies were qualitatively rationalized by the binding site models of the alpha 2-adrenoceptor. The benzylic methyl group of medetomidine or the naphthyl analog 2a was superimposable with the alpha-methyl group of (-)-alpha-methylnorepinephrine and fit into the proposed "methyl pocket" of the alpha 2-adrenoceptor defined by the residues Leu110, Leu169, Phe391, and Thr395.

Adrenergic alpha-Agonists↗