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Biomedical subjects

H Du

Publications and source records attributed to H Du.

At least 55 records · Page 3Linked to original sources
PubMed1999

[The design of an ultrasonics Doppler spectrum analysis system for fetal umbilical blood flow monitoring].

This paper explains the design of an Ultrasonics Doppler Spectrum Analysis System for fetal umbilical blood flow monitoring. The gist of discussion includes the sampling method of the Doppler signals and the algorithm of the blood flow parameters. Because the DSP chip is not used in the AD cardboard, the hardware design is simplified and the system works steadily. In the process of computing the blood flow paramenters, an autochange method of the amplitude threshold and a timeout control technology are used, so the algorithm has good adaptability and works very stable.

Algorithms↗
PubMed1999

[Use PCR technique to detect mitochondrial DNA in rat membranous labyrinth].

OBJECTIVE: To establish a sensitive and reliable method of mtDNA detection in the membranous labyrinth of rat inner ear. METHOD: A mtDNA segment of 601 bp containing the genes which encode ND1 subunit 16S rRNA in rat inner ear membranous labyrinth was detected by PCR method. The method is compared with Edris's mtDNA isolation method. RESULT: A 601 bp PCR product of mtDNA in the membranous labyrinth of rat ear inner can be obtained by the two methods. But the requirement of the sample quantities was different for the two methods. The extraction of mtDNA need two of inner ears membranous labyrinth to get reliably result by the method adapted from Seidman. However, the mtDNA extraction procedure from Edris's method need six of inner ears of the in order to get reliably result. CONCLUSION: The method of mtDNA isolation, amplification and detection membranous labyrinth of rat inner ear by PCR method adapted from seidman is more sersitive.

Animals↗
PubMed1999

[Study on the posterior vitreous detachment in patients with high myopia].

OBJECTIVE: To access the development of posterior vitreous detachment(PVD) related to age, axial length of the eye, vitreous changes as well as the myopic chorioretinal manifestation in patients with high myopia. METHODS: The vitreous status of 61 consecutive patients with high myopia(> or = -8.0 D and axial length > or = 26.0 mm) 110 eyes and 68 controls (< or = -3.0 D) 119 eyes were examined with biomicroscopic slit lamp and B-scan ultrasound. The myopic chorioretinal changes at the posterior pole were examined by indirect ophthalmoscope. The axial length of the eye was measured by A-scan ultrasound. The incidence of PVD was evaluated by Chi-square test. RESULTS: The incidence of PVD in patients with high myopia increased with age. Its overall incidence was significantly higher than that in the controls (78.2% vs 31.1%, P < 0.001). In addition, the onset of PVD in patients with high myopia occurred much earlier in age than that in the control group. Axial length was an important factor responsible for the development of PVD in patients with high myopia. The incidence of PVD and age were highly correlated (r = 0.9997, P < 0.05). Retinal detachment occurred in 5 eyes in the patients with high myopia. CONCLUSIONS: The results in the present study suggested that the incidence of PVD in patients with high myopia, increased with axial length of the eye and age. Vitreous changes are characteristic findings associated with high myopia, which was also involved in the development of PVD.

Adult↗
PubMed1999

Immunocytochemical study of cells in the vitreous of proliferative vitreoretretinopathy.

PURPOSE: To identify the cellular components of vitreous samples obtained during vitrectomy for proliferative vitreoretinopathy (PVR). METHODS: With the use of three intermediate filament (IF) proteins, vimentin, glial fibrillary acidic protein (GFAP), and cytokeratin (CK), cytocentrifuge slides of 14 fresh vitreous aspirates were detected with immunohistochemical technique. RESULTS: All the specimens contained epithelial-like proliferative cells with or without pigment and some membrane-like pieces. Immunocytochemical staining showed that 76.0-90.0% cells stained for CK, 17.4-29.6% cells expressed GFAP, and 80.1-91.0% cells were positive for vimentin. CONCLUSIONS: Majority of cells in the vitreous samples originated from retinal pigment epithelial cells (RPE) and glial cells in PVR. Expression of IF proteins may be determined by tissue of origin and local microenvironment.

Adolescent↗
PubMed1998

trp RNA-binding attenuation protein-mediated long distance RNA refolding regulates translation of trpE in Bacillus subtilis.

Expression of the trpEDCFBA operon is regulated at both the transcriptional and translational levels by the trp RNA-binding attenuation protein (TRAP) of Bacillus subtilis. When cells contain sufficient levels of tryptophan to activate TRAP, the protein binds to trp operon transcripts as they are being synthesized, most often causing transcription termination. However, termination is never 100% efficient, and transcripts that escape termination are subject to translational control. We determined that TRAP-mediated translational control of trpE can occur via a novel RNA conformational switch mechanism. When TRAP binds to the 5'-untranslated leader segment of a trp operon read-through transcript, it can disrupt a large secondary structure containing a portion of the TRAP binding target. This promotes refolding of the RNA such that the trpE Shine-Dalgarno sequence, located more than 100 nucleotides downstream from the TRAP binding site, becomes sequestered in a stable RNA hairpin. Results from cell-free translation, ribosome toeprint, and RNA structure mapping experiments demonstrate that formation of this structure reduces TrpE synthesis by blocking ribosome access to the trpE ribosome binding site. The role of the Shine-Dalgarno blocking hairpin in controlling translation of trpE was confirmed by examining the effect of multiple nucleotide substitutions that abolish the structure without altering the Shine-Dalgarno sequence itself. The possibility of protein-mediated RNA refolding as a general mechanism in controlling gene expression is discussed.

Bacillus subtilis↗
PubMed1998

Mouse lysosomal acid lipase: characterization of the gene and analysis of promoter activity.

Lysosomal acid lipase (LAL) is required for the hydrolysis of intracellular cholesteryl esters and triglycerides that are delivered to lysosomes by low density lipoprotein (LDL) receptor-mediated endocytosis. To understand that the expression of LAL mRNA and protein is tissue and cell specifically regulated in mice, genomic clones for the mouse lysosomal acid lipase (mLAL) gene were isolated and characterized. The 6.8 kb of the mLAL gene 5'-flanking region was sequenced. Comparisons of mouse and human LAL genes organization revealed identical intron/exon boundaries, except for intron 1 of the mouse gene, and identical exonic length of exons 3-9. The transcription start sites and exon 1 of mLAL were characterized by 5'-RACE-PCR and S1 nuclease mapping. Transfection of 5' flanking deletions of mLAL luciferase reporter gene construct identified positive and negative regulatory elements that varied with cell type. Transfection of three progressively smaller pieces of intron 1 inserted into an SV40 promoter and luciferase reporter gene revealed an enhancer-like activity in intron 1 that is also cell type specific. These studies provide insight into the basis for regulation of this critical enzyme in lipid metabolism.

Animals↗
PubMed1998

Molecular and enzymatic analyses of lysosomal acid lipase in cholesteryl ester storage disease.

Human lysosomal acid lipase (hLAL) is essential for the hydrolysis of cholesteryl esters and triglycerides in the lysosome. Defective hLAL activity leads to two autosomal recessive traits, Wolman disease (WD) or cholesteryl ester storage disease (CESD). Phenotypically, WD has accumulation of both triglycerides and cholesteryl esters, while CESD has mainly elevated cholesteryl esters. We characterized mutations in the hLAL gene from two CESD siblings. By reverse transcriptase-PCR (RT-PCR) and cDNA cloning and sequencing, we identified homozygous deletion mutations of nucleotides 863 to 934, in the hLAL transcript. Normal levels of LAL mRNA were detected. The deletion in mRNA is due to a G to A transition in the last nucleotide of exon 8 of the hLAL gene, a splice junction mutation (E8SJM) that resulted in exon skipping, and a predicted in-frame deletion of the 24 amino acids. [35S]Met metabolic labeling studies in fibroblasts showed a low level of E8SJM LAL ( approximately 38%) that was highly unstable. Heterologous expression of E8SJM LAL in insect cells gave an LAL with low catalytic activity toward cholesteryl oleate and triolein. The effects of this mutation are complex with the production of decreased amounts of an unstable LAL that is catalytically defective. The results suggest that E8SJM leads to essentially a null allele and that the differences in WD and CESD phenotype involve other factors.

Animals↗
PubMed1998

Targeted disruption of the mouse lysosomal acid lipase gene: long-term survival with massive cholesteryl ester and triglyceride storage.

Lysosomal acid lipase (LAL) is essential for the hydrolysis of the triglycerides and cholesteryl esters in lysosomes. Its deficiency produces two phenotypes, a severe infantile-onset variant, Wolman disease (WD), and a later onset variant, cholesteryl ester storage disease (CESD). A mouse model with a LAL null mutation was produced by targeting disruption of the mouse gene. Homozygote knockout mice (lal -/lal-) produce no LAL mRNA, protein or enzyme activity. The lal-/lal- mice are born in Mendelian ratios, are normal appearing at birth, and follow normal development into adulthood. However, massive accumulation of triglycerides and cholesteryl esters occurs in several organs. By 21 days, the liver develops a yellow-orange color and is approximately 1.5-2.0x larger than normal. The accumulated cholesteryl esters and triglycerides are approximately 30-fold greater than normal. The lal+/lal- mice have approximately 50% of normal LAL activity and do not show lipid accumulation. Male and female lal-/lal- mice are fertile and can be bred to produce progeny. This mouse model is a phenotypic model of human CESD, and a biochemical and histopathologic mimic of human WD. The lal-/lal- mice provide a model to determine the role of LAL in lipid metabolism and the pathogenesis of its deficiency states.

Animals↗
PubMed1998

Activation of the tobacco SIP kinase by both a cell wall-derived carbohydrate elicitor and purified proteinaceous elicitins from Phytophthora spp.

Two purified proteinaceous fungal elicitors, parasiticein (an alpha elicitin) and cryptogein (a beta elicitin), as well as a fungal cell wall-derived carbohydrate elicitor all rapidly activated a 48-kD kinase in tobacco suspension cells. The maximum activation of this kinase paralleled or preceded medium alkalization and activation of the defense gene phenylalanine ammonia-lyase (PAL). In addition, the two elicitins, which also induced hypersensitive cell death, activated a 44- and a 40-kD kinase with delayed kinetics. By contrast, the cell wall-derived elicitor only weakly activated the 44-kD kinase and failed to activate the 40-kD kinase. The size and substrate preference of the 48-kD kinase are reminiscent of the recently purified and cloned salicylic acid-induced protein (SIP) kinase, which is a member of the mitogen-activated protein kinase family. Antibodies raised against a peptide corresponding to the unique N terminus of SIP kinase immunoreacted with the 48-kD kinase activated by all three elicitors from Phytophthora spp. In addition, the cell wall elicitor and the salicylic acid-activated 48-kD kinase copurified through several chromatography steps and comigrated on two-dimensional gels. Based on these results, all three fungal elicitors appear to activate the SIP kinase. In addition, inhibition of SIP kinase activation by kinase inhibitors correlated with the suppression of cell wall elicitor-induced medium alkalization and PAL gene activation, suggesting a regulatory function for the SIP kinase in these defense responses.

Algal Proteins↗
PubMed1998

The epidemiology study of hyperuricemia and gout in a community population of Huangpu District in Shanghai.

OBJECTIVE: To investigate the prevalence of hyperuricemia and gout in a community population of Huangpu District in Shanghai. METHODS: In the target community, 2037 dwellers were interviewed with relevan questionnares from house to house. According to even house number 1017 blood samples were taken for serum uric acid (SUA) levels measured with the uricase-peroxidase enzymatic method. RESULTS: The prevalence of hyperuricemia was 14.2% in men (SUA > 70 mg/L, 62 cases), 7.1% in women (SUA > 60 mg/L, 41 cases), 10.1% in both sexes. Seven gout patients were all men. The prevalence of gout in 2037 dwellers in Huangpu District was 0.77% in men and 0.34% in both sexes. CONCLUSIONS: The mean SUA level in each age group in this survey was much higher than that of a previous study 1 carried out in Shanghai, Beijing and Guangzhou in 1980 (P < 0.05). And the prevalence of hyperuricemia was increased rapidly (in men: from 1.4% in the survey of 1980 to 14.2% in our survey; in women: from 1.3% in the survey of 1980 to 7.1% in our survey). Compared with Idonesia data in 1992, the prevalence of hyperuricemia and gout in our survey was lower than that in Indonesia (P < 0.05), which suggests that racial and genetic predispositions are key causative factors.

Adolescent↗
PubMed1998

[Chemical constituents of Cichorium intybus L].

OBJECTIVE: To investigate the chemical constituents of Cichorium intybus. METHOD: Constituents were separated by means of solvent extraction and chromatography on silical gel. The structures were established by spectral analysis and chemical evidence. RESULT: Seven compounds were isolated from the roots of C. intybus and four of them were identified as alpha-amyrin, taraxerone, baurenyl acetate and beta-sitosterol. CONCLUSION: All the four compounds were obtained from the plant for the first time.

Cichorium intybus↗
PubMed1998

EMG study in the diagnosis and differential diagnosis of lipid storage myopathy.

Clinical, electromyographic and pathological features were studied in 18 patients with lipid storage myopathy (group I) and 18 patients with polymyositis and dermatomyositis (group II). The results showed a remarkable lower spontaneous activity(SA) incidence (14%) in group I than that (55%) in group II; 46% and 34% short-duration motor unit potentials(MUAPs) with polyphasic potentials and 74% and 71% short-duration MUAPs without polyphasic potentials respectively; the percentages of increased polyphasic MUAPs were same in the two groups. The reduced or pathologic interference patterns accounted for 61% in the group I and 50% in group II. Increased CPK, LDH and HBD were also found in both of them. It is suggested that the lipid storage myopathy may be diagnosed when patients have muscle weakness and myalgia with short-duration and low-amplitude and polyphasic MUAPs without or with occasional spontaneous activities, and increased CPK, LDH and HBD.

Adolescent↗
PubMed1998

[The design of the ultrasonic Doppler spectrum analyze system for the naval blood stream of fetus].

The design of the Ultrasonic Doppler Specturm Analyze System for the naval blood stream of the fetus is discussed, mainly on the sampling method of the Doppler signals and the algorithm of the blood stream parameters. Because the DSP chip is not used in the AD cardboard, the hardware design is not only simplified, but also works more steady. In the process of the blood stream parameters, an autochange method of the amplitude threshold and a time-out control technology are used, so the algorithm has good adaptation and works very stable.

Algorithms↗
PubMed1997

Cloning of an inr- and E-box-binding protein, TFII-I, that interacts physically and functionally with USF1.

The transcription factor TFII-I has been shown to bind independently to two distinct promoter elements, a pyrimidine-rich initiator (Inr) and a recognition site (E-box) for upstream stimulatory factor 1 (USF1), and to stimulate USF1 binding to both of these sites. Here we describe the isolation of a cDNA encoding TFII-I and demonstrate that the corresponding 120 kDa polypeptide, when expressed ectopically, is capable of binding to both Inr and E-box elements. The primary structure of TFII-I reveals novel features that include six directly repeated 90 residue motifs that each possess a potential helix-loop/span-helix homology. These unique structural features suggest that TFII-I may have the capacity for multiple protein-protein and, potentially, multiple protein-DNA interactions. Consistent with this hypothesis and with previous in vitro studies, we further demonstrate that ectopic TFII-I and USF1 can act synergistically, and in some cases independently, to activate transcription in vivo through both Inr and the E-box elements of the adenovirus major late promoter. We also describe domains of USF1 that are necessary for its independent and synergistic activation functions.

Adenoviridae↗
PubMed1997

Cloning, chromosome localization, expression, and characterization of an Src homology 2 and pleckstrin homology domain-containing insulin receptor binding protein hGrb10gamma.

hGrb10alpha (previously named Grb-IR) is a Src-homology 2 domain-containing protein that binds with high affinity to the tyrosine-phosphorylated insulin receptor and insulin-like growth factor-1 receptor. At least two isoforms of human Grb10, (hGrb10alpha and hGrb10beta), which differ in the pleckstrin homology (PH) domain and the N-terminal sequence, have previously been identified in insulin target tissues such as human skeletal muscle and fat cells. Here we report the cloning of the third isoform of the hGrb10 family (hGrb10gamma) from human skeletal muscle and its localization to human chromosome 7. We have also determined the human chromosome localization of Grb7 to 17q21-q22 and Grb14 to chromosome 2. hGrb10gamma contains an intact PH domain and an N-terminal sequence that is present in hGrb10alpha but absent in hGrb10beta. RNase protection assays and Western blot analysis showed that hGrb10alpha and hGrb10gamma are differentially expressed in insulin target cells including skeletal muscle, liver, and adipocyte cells. hGrb10gamma is also expressed in HeLa cells and various breast cancer cell lines. The protein bound with high affinity to the insulin receptor in cells, and the interaction was dependent on the tyrosine phosphorylation of the receptor. hGrb10gamma also underwent insulin-stimulated membrane translocation and serine phosphorylation. hGrb10gamma phosphorylation was inhibited by PD98059, a specific inhibitor of mitogen-activated protein kinase kinase, and wortmannin, a specific inhibitor of phosphatidylinositol 3-kinase. Taken together, our data suggest that hGrb10 isoforms are potential downstream signaling components of the insulin receptor tyrosine kinase and that the PH domain may play an important role in the involvement of these isoforms in signal transduction pathways initiated by insulin and other growth factors.

Amino Acid Sequence↗
PubMed1997

Liver-directed gene transfer in non-human primates.

To develop a primate model for liver-directed gene therapy, we studied several gene transfer vehicles and routes in eight rhesus monkeys (Macaca mulatta). For this purpose, we used first-generation, replication-deficient adenoviral vectors carrying the Escherichia coli lacZ gene (Ad.CMVlacZ) or a lacZ-containing plasmid (pCMV beta) with lipofectamine for transfection. The reporter gene construct was infused into either the portal vasculature, common bile duct, or saphenous vein. Adenovirus-mediated gene transfer via the portal vein resulted in expression of lacZ in over 70% of hepatocytes by days 3-7, but was accompanied by acute hepatitis. Adenovirus-mediated gene transfer via the common bile duct resulted in lacZ expression in less than 10% of hepatocytes and was accompanied by portal inflammation. The animals mounted a significant immune response, as demonstrated by adenoviral antigen-induced T-cell proliferation and production of neutralizing anti-adenovirus antibodies and antibodies to E. coli beta-galactosidase (beta-Gal). Activation of the immune response was associated with rapid decrease of the reporter gene by days 13-21. Lipofectamine-mediated gene transfer was inefficient, and no lacZ expression in the liver was detected. To limit the host immune response, 4 animals were immunosuppressed by cyclophosphamide/prednisone and then infused with the Ad.CMVlacZ via the portal vein or the saphenous vein. The monkeys showed sustained expression of lacZ for up to 35 days with no evidence of inflammation. The primates transduced via the saphenous vein showed a level of beta-Gal expression in the liver similar to that of the portal vein-infused animals. In conclusion, adenovirus-mediated gene transfer to non-human primate livers via the portal vein or saphenous vein is efficient, but it results in transient expression and is accompanied by an immune response to both vector and transgene products and acute hepatitis, whereas lipofectamine-mediated transfer is inefficient. Manipulation of the host immune response may expand potential applications of adenoviral vectors for liver-directed gene transfer.

Adenoviridae↗