[Solid phase extraction--HPLC in determination of coumatetralyl in urine].
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Biomedical subjects
Publications and source records attributed to Li Wang.
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Using THz time-domain spectroscopy, the home-grown organic crystal DAST and widely used high-efficient ZnTe were characterized as THz emitters. Bolometer was also used to measure the absolute energy of THz radiation from DAST and ZnTe. The refractive index and absorption coefficient of DAST were measured too. It is concluded that the home-grown DAST is a kind of high-efficiency THz material, and could become a good THz emitter and detector after the improvement in growing process.
OBJECTIVE: To study on the various proportions of Radix Sophorae Flavescentis, Cortex Phellodendri, Fructus Cnidii and pericarp of Zanthoxylum bungeanum Maxim in the formulas, whose antimicrobial effects on E. coli, S. aureus, P. aeruginosa and C. albicans under different pH values were compared in vitro. METHODS: According to Chinese ancient proved recipe, the K-B method and plate diluting method were adopted to measure antimicrobial activity, and orthogonal design to ascertain the herbal formula in vitro. Finally, murine models were established to test the antimicrobial activity in vivo through vaginal membrane irritancy experiment, negative rate of pathogeny and pathological grade of vaginal membrane. RESULTS: The results suggested that formulas with different proportions of the herbs had diverse antimicrobial activities, and the effect was shown to be most obvious when one milliliter drug contains 100 microl Fructus Cnidii-pericarp of Zanthoxylum bungeanum (2:1) co-extracted volatile oil and 50 microl Radix Sophorae Flavescentis and Cortex Phenodendri ethanol extraction respectively under pH6. The antimicrobial effect of the formula, which hardly had any membrane irritancy, was better than Jie Eryin in vitro and vivo. CONCLUSION: The fromula has few components and better effect, and adaptation to the pH value of vaginitis. It is a promising alternative for gynecological diseases.
A large-scale lambda-DNA network on a mica surface was successfully fabricated with a simple method. Silver nanoparticles capped with the cationic surfactant cetyltrimethylammonium bromide (CTAB) were self-assembled onto a two-dimensional DNA network template by electrostatic interaction and formed nanoporous silver films, which can be used as active surface-enhanced raman scattering (SERS) substrates. Two probe molecules, Rhodamine 6G (R6G) and 4-aminothiophenol (4-ATP), were studied on these substrates with very low concentrations, and great enhancement factors for R6G (0.21 x 10(10)-4.09 x 10(11)) and 4-ATP (approximately 1.70 x 10(5)) were observed. It was found that the enhancement ability was affected by the DNA concentration and the electrostatic absorption time of the CTAB-stabilized silver nanoparticles on the DNA strands. These SERS substrates formed by the self-assembly of silver nanoparticles on DNA network also show good stability and reproducibility in our experiments.
Ischemic preconditioning confers powerful protection against myocardial infarction through pre-emptive activation of survival signaling pathways, but it remains difficult to apply to patients with ischemic heart disease, and its effects are transient. Promoting a sustained activation of preconditioning mechanisms in vivo would represent a novel approach of cardioprotection. We tested the role of the protein H11 kinase (H11K), which accumulates by 4- to 6-fold in myocardium of patients with chronic ischemic heart disease and in experimental models of ischemia. This increased expression was quantitatively reproduced in cardiac myocytes using a transgenic (TG) mouse model. After 45 minutes of coronary artery occlusion and reperfusion, hearts from TG mice showed an 82+/-5% reduction in infarct size compared with wild-type (WT), which was similar to the 84+/-4% reduction of infarct size observed in WT after a protocol of ischemic preconditioning. Hearts from TG mice showed significant activation of survival kinases participating in preconditioning, including Akt and the 5'AMP-activated protein kinase (AMPK). H11K directly binds to both Akt and AMPK and promotes their nuclear translocation and their association in a multiprotein complex, which results in a stimulation of survival mechanisms in cytosol and nucleus, including inhibition of proapoptotic effectors (glycogen synthase kinase-3beta, Bad, and Foxo), activation of antiapoptotic effectors (protein kinase Cepsilon, endothelial and inducible NO synthase isoforms, and heat shock protein 70), increased expression of the hypoxia-inducible factor-1alpha, and genomic switch to glucose utilization. Therefore, activation of survival pathways by H11K preemptively triggers the antiapoptotic and metabolic response to ischemia and is sufficient to confer cardioprotection in vivo equally potent to preconditioning.
OBJECTIVE: To investigate the mechanism of the different biological impacts of HBx3'-40, an engineered deletion mutant lacking the last 40 C-terminal amino acids. METHOD: Human hepatocellular cells of the line Huh7 were transfected with HBx3'-40 or wtHBx (wild type HBx) constructs. An oligo cDNA microarray containing 21074 human genes and Ests was utilized to examine the different gene expression between HBx3'-40 and paired control wtHBx cells. A series of methods, including immobilized pH gradient two-dimensional polyacrylamide gel electrophoresis (2-DE), silver staining, and PDQuest 2-D analysis software were used to analyze the differential protein-spots between HBx3'-40 and wtHBx cells. Selected differential protein-spots were identified with peptide mass fingerprinting based on matrix-assisted laser desorption/ionization time-of-fight mass spectrometry (MALDI-TOF-MS) and database searching. RESULTS: 165 (0.82%) candidate genes showing aberrant expression under HBx3'-40 induction were identified, of which 144 were up-regulated while 21 were down-regulated. Compared with wtHBx group, there were 135 +/- 13 differently protein spots in HBx3'-40 group by 2-DE. Of them, 7 significantly different protein spots were identified using mass spectrometry and computer matching with protein database. Most of the differently expressed genes and proteins were involved in transcription, oncogenes and tumor suppressor genes or protein, cell adhesion, signal transduction pathways, metabolisms, etc. CONCLUSION: The mutant HBx affects cell genes and proteins involved in various processes, especially in regulation of liver metabolism.
The PIK3CA gene encoding the p110alpha subunit of Class IA phosphatidylinositol 3-kinases (PI3Ks) is frequently mutated in human tumors. Mutations in the PIK3CB gene encoding p110beta, the only other widely expressed Class IA PI3K, have not been reported. We compared the biochemical activity and transforming potential of mutant forms of p110alpha and p110beta in a human mammary epithelial cell system. The two most common tumor-derived alleles of p110alpha, H1047R and E545K, potently activated PI3K signaling. Human mammary epithelial cells expressing these alleles grew efficiently in soft agar and as orthotopic tumors in nude mice. We also examined a third class of mutations in p110alpha, those in the p85-binding domain. A representative tumor-derived p85-binding-domain mutant R38H showed modestly reduced p85 binding and weakly activated PI3K/Akt signaling. In contrast, a deletion mutant lacking the entire p85-binding domain efficiently activated PI3K signaling. When we constructed in p110beta a mutation homologous to the E545K allele of p110alpha, the resulting p110beta mutant was only weakly activated and allowed minimal soft-agar growth. However, a gene fusion of p110beta with the membrane anchor from c-Src was highly active and transforming in both soft-agar and orthotopic nude mouse assays. Thus, although introduction of activating mutations from p110alpha at the corresponding sites in p110beta failed to render the enzyme oncogenic in human cells, the possibility remains that other mutations might activate the beta isoform.
This study determined: (a) if acupuncture stimulation at a traditional site might modulate ongoing EEG as compared with stimulation of a control site; (b) if high-frequency vs. low-frequency stimulation could exert differential effects of acupuncture; (c) if the observed effects of acupuncture were specific to certain EEG bands; and (d) if the acupuncture effect could be isolated at a specific scalp field, with its putative underlying intracranial source. Twelve healthy male volunteers (age range 22-35) participated in two experimental sessions separated by 1 week, which involved transcutaneous acupoint stimulation at selected acupoint (Li 4, HeGu) vs. a mock point at the fourth interosseous muscle area on the left hand in high (HF: 100 Hz) vs. low-frequency (LF: 2 Hz) stimulation by counter-balanced order. 124-ch EEG data were used to analyze the Delta, Theta, Alpha-1, Alpha-2, Beta, and Gamma bands. The absolute EEG powers (muv2) at focal maxima across three stages (baseline, stimulation, post) were examined by two-way (condition, stage) repeated measures ANOVA. The activity of the Theta power significantly decreased (P = 0.02), compared with control during HF but not LF stimulation at acupoint stimulation, however, there was no study effect at the mock point. A decreased Theta EEG power was prominent at the frontal midline sites (FCz, Fz) and the contralateral right hemisphere front site (FCC2h). In contrast, the Theta power of low-frequency stimulation showed an increase from the baseline as those in both controlled mock point stimulations. The observed high-frequency acupoint stimulation effects of Theta EEG were only present during, but not after, simulation. The topographic Theta activity was tentatively identified to originate from the intracranial current source in cingulate cortex, likely ACC. It is likely that short-term cortical plasticity occurs during high-frequency but not low-frequency stimulation at the HeGu point, but not mock point. We suggest that HeGu acupuncture stimulation modulates limbic cingulum by a frequency modulation mode, which then may damp nociceptive processing in the brain.
OBJECTIVE: To investigate the clinical features of early onset narcolepsy. METHODS: The clinical data of 105 consecutive patients with narcolepsy, 63 of which with an onset age of 9.7 +/- 3.1 on average and 42 with an onset age of 22.8 +/- 9.3 on average. Interrogation, physical examination, CT, and MRI were carried out. Polysomnography was conducted. Then the data were compared between these 2 groups. RESULTS: All 105 patients showed daytime sleepiness. The incidence rate of cataplexy was 92% in the early onset group, significantly higher than that of the late onset group (P = 0.023). There were no significant differences in the rates of sleep paralysis, hypnagogic hallucination, and disturbed nocturnal sleep. Multiple sleep latency test showed that the mean sleep latency of the early-onset group was 4.5 +/- 4.0 min, significantly shorter than that of the late onset group (7.0 +/- 5.7 min, P = 0.018); the REM sleep latency of the early onset group was 3.4 +/- 3.2 min, significantly shorter than that of the late onset group (4.8 +/- 2.2 min, P = 0.02). The number of REM sleep of the early onset group was 3.4 +/- 2.0, significantly more than that of the late onset group (2.5 +/- 1.9, P = 0.09). The apnea and hypopnea index of the late onset group was 9.2 +/- 16.5, significantly higher than that of the early onset group (1.9 +/- 6.3, P = 0.009). The 3 cases of narcolepsy with family history were all cases of early onset narcolepsy. CONCLUSION: Early onset narcolepsy patients have more severe daytime sleepiness and higher rate of cataplexy. The pathogenesis of early onset narcolepsy may be more closely associated with genetic factors.
Results of gene expression microarray and quantitative PCR studies have suggested abnormalities in the expression of myelin-related genes including myelin-associated glycoprotein (MAG) in schizophrenic patients. Research provides strong evidence for oligodendrocyte dysfunction in schizophrenics. In order to further assess the role of MAG in schizophrenia, we examined four single nucleotide polymorphisms (SNPs), namely rs2301600, rs3746248, rs720309 and rs720308, of this gene in Chinese schizophrenic patients (n=470) and healthy controls (n=470). The distribution of rs720309 T/A genotypes showed a strong association with schizophrenia (chi(2)=14.58, d.f.=2, P=0.0008). A haplotype constructed of rs720309-rs720308 also revealed a significant association with schizophrenia (chi(2)=11.914, d.f.=3, P=0.0084). Our findings of a significant associations between schizophrenia and the MAG gene suggest that this gene may be involved in susceptibility to schizophrenia in the Chinese Han population.
To develop CpG oligodeoxynucleotides (CpG ODNs) based therapy for prevention and treatment of severe acute respiratory syndrome (SARS), we selected a novel CpG ODN (BW001), which displays B-type CpG ODN structure feature at the 5' and A-type CpG ODN structure feature at the 3', and tested for its anti-SARS-CoV activity. We found that the supernatants of human PBMCs stimulated by BW001 significantly protected Vero cells from SARS-CoV infection. BW001 could stimulate human PBMCs and pDCs to secrete high level of IFN-alpha and promote human PBMCs and B cells to proliferate. Furthermore, we demonstrated that BW001 could activate CD19+ B cells and CD56+ NK cells in human PBMCs. In addition, BW001 could enhance NK cytotoxicity and IFN-gamma secretion in human PBMCs. Together, BW001 represents a novel type of CpG ODN and may have potential for the development of treatment and prevention for SARS as well as other viral associated diseases.
Calcyclin-binding protein (Siah-1-Interacting Protein, CacyBP/SIP), is a calcium signaling protein involved in the degradation of beta-catenin, however, little is known about its role in reproductive biology. The present study was to character its temporospatial expression pattern and regulation in mouse uterus and to investigate whether it plays a role in the regulation of normal endometrial events. While prominently expressed in both luminal and glandular epithelia, CacyBP underwent dynamic changes during early pregnancy. CacyBP expression was observed weakly from days 1-4. An intense accumulation in luminal and glandular epithelia as well as decidua surrounding the embryo at later stages (days 5-7) was observed. Most notably, CacyBP accumulation in trophoblast was pronounced at day 7. Using ovariectomized and pseudopregnant mice, we found that progesterone (P(4)) and 17beta-estradiol (E(2)) led to increased expression of CacyBP gene and this could be abolished by Ru486 and tamoxifen, respectively. Antisense oligonucleotides (ODNs) against CacyBP significantly inhibited cultured endometrial stromal cells' (ESCs) apoptosis induced by UV irradiation. Injection of antisense ODNs into mouse uterine horn severely impaired the number of implanted blastocysts. Taken together, our results suggested that CacyBP expression was positively regulated by P(4) and E(2). CacyBP may be involved in the regulation of endometrial cell apoptosis during early pregnancy and play an important role in mouse endometrial events such as pregrancy establishment.
Cholesterol homeostasis in mammals involves pathways for biosynthesis, cellular uptake, and hepatic conversion to bile acids. Key genes for all three pathways are regulated by negative feedback control. Uptake and biosynthesis are directly regulated by cholesterol through its inhibition of the proteolytic activation of the sterol regulatory element binding proteins. The conversion of cholesterol into bile acids in the liver is regulated through the bile acid-dependent induction of the negatively acting small heterodimer partner nuclear receptor. In this report, we have shown that the small heterodimer partner also directly regulates cholesterol biosynthesis through inhibition of 3-hydroxy-3-methylglutaryl coenzyme A reductase but has no effect on low density lipoprotein receptor expression. This has significant metabolic significance, as it provides both a mechanism to independently regulate cholesterol synthesis from uptake (an essential regulatory feature known to occur in vivo) and a pathway for direct regulation of cholesterol biosynthesis by bile acids. This latter feature ensures that the early phase of bile acid synthesis (pre-cholesterol) is in metabolic communication with the later stages of the pathway to properly regulate whole pathway flux. This highlights an important regulatory feature that is shared with other key branched, multienzyme pathways, such as glycolysis, where pathway outflow through pyruvate kinase is regulated by the concentration of a key early intermediate, fructose 1,6-bisphosphate.
PURPOSE: To identify genetic defects associated with nuclear golden crystal autosomal dominant congenital cataract (ADCC) in a Chinese pedigree in the north of China. METHODS: Clinical data were collected and the phenotype of the affected members in this family was recorded by slit lamp photography. Genomic DNA was isolated from peripheral blood. Linkage analyses excluded all known loci except that in 2q33-q35. Mutation analysis of CRYGs was carried by direct sequencing of the PCR products. RESULTS: Sequencing of the coding regions of CRYGA, CRYGB, CRYGC, and CRYGD showed the presence of a heterozygous C>A transversion at nt109 of the coding sequence (R36S) in exon 2 of CRYGD, which co-segregated with the affected members. CONCLUSIONS: The R36S mutation in CRYGD identified in this Chinese family caused a nuclear golden crystal cataract phenotype not described before. This finding is an additional indication that there may be phenotypic heterogeneity of cataract, especially in different races.
AIM: To investigate the effect of six bile salts: glycocholate (GC), glycochenodeoxycholate (GCDC), glycodeoxycholate (GDC), taurocholate (TC), taurochenodeoxycholate (TCDC), taurodeoxycholate (TDC), and their mixture on cultured human normal esophageal mucosal epithelial cells. METHODS: Human normal esophageal mucosal epithelial cells were cultured with serum-free keratinocyte medium.3-[4,5-Dimethylthiaolyl]-2,5-diphenyl-tetrazolium bromide assay was applied to the detection of cell proliferation. Apoptotic morphology was observed by phase-contrast video microscopy and terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay. Sub-G1 DNA fragmentations and early apoptotic cells were assayed by flow cytometry (FCM) with propidium iodide (PI) staining and annexin V-FITC conjugated with PI staining. Apoptotic DNA ladders on agarose gel electrophoresis were observed. RESULTS: Except for GC, GCDC, GDC, TC, TCDC, TDC and their mixture could initiate growth inhibition of esophageal mucosal epithelial cells in a dose- and time-dependent manner. TUNEL and FCM assays demonstrated that the bile salts at 500 mumol/L and their mixture at 1 500 micromol/L induced apoptosis except for GC. The percentage of sub-G1 detected by FCM with PI staining was 83.5% in cells treated with 500 micromol/L TC for 2 h, and 19.8%, 20.4%, 25.6%, 13.5%, and 75.8% in cells treated with 500 micromol/L GCDC, TCDC, GDC, TDC, and 1 500 micromol/L mixture for 24 h, respectively, which were higher than that of the control (1.5%). The percentage was 1.4% in cells with 500 micromol/L GC for 24 h. DNA ladders on agarose gel electrophoresis were seen in cells treated with 500 micromol/L TC for 2 h and 1 500 micromol/L mixture for 24 h. CONCLUSION: All GCDC, GDC, TC, TCDC, TDC and their mixture can inhibit growth and induce apoptosis of cultured human normal esophageal mucosal epithelial cells, but GC is well tolerated by the cells.
Alpha-latrotoxin, a potent stimulator of exocytosis from neurons and neuroendocrine cells, has been studied intensively, but the mechanisms of its actions are poorly understood. Here, we developed a new method to generate active recombinant alpha-latrotoxin and conducted a structure/function analysis of the toxin in stimulating Ca2+-dependent exocytosis. alpha-Latrotoxin consists of a conserved N-terminal domain and C-terminal ankyrin-like repeats. After cleavage of an N-terminally fused purification tag of glutathione S-transferase (GST), the recombinant toxin strongly stimulated exocytosis, whereas the GST-fused toxin was much less potent. The GST-fused toxin bound to the receptors [neurexin 1alpha; CL1 (CIRL/latrophilin 1)] as efficiently as did the GST-cleaved toxin but was much less effective in inserting into the plasma membrane and inducing cation conductance. The toxin with deletion of the last two ankyrin-like repeats still bound the receptors but could neither stimulate exocytosis nor induce cation conductance efficiently. The abilities of the mutated toxins to stimulate exocytosis correlated well with their abilities to induce cation conductance, but not their binding to the receptors. Our results indicate that (1) C-terminal ankyrin-like repeats and a free (unfused) N terminus are both required for the toxin to form pores, which is essential for Ca2+-dependent exocytosis, and (2) receptor binding alone is not sufficient to stimulate Ca2+-dependent exocytosis.
Tyroserleutide (YSL) is a type of active, low molecular weight polypeptide, comprised of three amino acids, which has antitumor effects. YSL has various advantages over the other bioactive peptides such as its low molecular weight, simple construction, nonimmunogenicity, specificity, few side effects, and ease of synthesis. However, the biological activities contributing to it's antitumor effects are not yet known. We studied the effects of YSL on the in vitro cytotoxic activity of BALB/c mice peritoneal macrophages (PEMphi) against the target tumor cell lines BEL-7402 and B16-F10. We also measured the concentrations of interleukin (IL)-1beta, tumor necrosis factor (TNF)-alpha, and nitric oxide (NO) produced by YSL-activated Mphi, and we determined the concentrations of IL-1beta and NO secreted by YSL-activated murine macrophage RAW264.7 cells. YSL activated Mphi in vitro, inhibited BEL-7402 proliferation, enhanced PEMphi antitumor effects, and stimulated IL-1beta, TNF-alpha, and NO production by RAW264.7 cells. These data suggest that YSL activates the monocyte-macrophage system, which enhances Mphi antitumor effects against BEL-7402 and B16-F10 cells and stimulates the secretion by Mphi of cytotoxic effectors such as IL-1beta, TNF-alpha, and NO.
Novel necklace-like polymer gel electrolytes containing latent chemically cross-linked gel electrolyte precursors were prepared for quasi-solid dye sensitized solar cells with a highest efficiency of 7.72% and an active area of 0.25 cm2 under AM1.5 condition at 100 mW cm(-2) irradiation.