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Biomedical subjects

S Wu

Publications and source records attributed to S Wu.

At least 415 records · Page 23Linked to original sources

[The phenoloxidase test and its application to identify Cryptococcus neoformans with various biological characteristics].

Cryptococcus neoformans strains with various biological characteristics were examinated the phenoloxidase activity on the caffeic acid cornmeal agar (CACA). Other medically important yeasts were also included in this research. Firstly, thirteen reference strains of Cryptococcus neoformans were confirmly found positive phenoloxidase production on the CACA medium. Then, in total 150 isolates of yeasts, forty three Cryptococcus neoformans strains were phenoloxidase positive, other than that 107 other yeast strains were all negative. It is suggested that Cryptococcus neoformans strains with different biological features specifically and uniquely produce phenoloxidase, thus the phenoloxidase test can be applied as a useful tool to identify this pathogenic yeast. Our results also show that the culture on the CACA medium is an effective method to test the phenoloxidase activity within 72 hours. It is further approved that the urease test will not be able to be a screening test for Cryptococcus neoformans in the clinical laboratory, but it is still valuable to identify urease negative Cryptococcus neoformans when the test is used in combination with the phenoloxidase test.

Candida↗

[Genetic diagnosis of a subtype diabetes mellitus with mitochondrial tRNA Leu(UUR) gene mutation].

The A-G point mutation of mitochondrial tRNALeu(UUR) gene at nucleotide 3243 caused clinical NIDDM in combination with or without deafness and was inherited through maternal transmission. An ApaI restriction site was resulted by this mutation, which could be detected by polymerase chain reaction (PCR) with ApaI digestion and be used in clinical genetic diagnosis. The first Chinese family with this mutation found through our screening of NIDDM patients was reported. Genetic diagnosis was done in 12 of 15 members in this three-generation pedigree. Positive results (present of ApaI restriction site) were observed in all 5 NIDDM patients (four with deafness). In 6 of 7 non-diabetic members, negative genetic diagnosis was obtained. The presymptomatic diagnosis of this disease was made in a 13-year old non-diabetic boy with positive genetic diagnosis in conjunction with the maternal segregation pattern of the mutation in this family. The finding of this disease in Chinese provides firm evidence of genetic heterogeneity in Chinese NIDDM. The techniques of the genetic diagnosis of this disease can be used in clinical laboratory, which indicates that in the diagnosis of NIDDN, the molecular etiologic level can be reached in daily clinical practice.

Adolescent↗

[Clinical observation and study of blood phosphate concentration changes in postoperative patients].

Blood phosphate concentration of 257 postoperative patients in the ICU was determined through the molybdenum-blue developing method. The results showed that 136 cases had hypophosphatemia (blood phosphate concentration < 0.8mmol/L), bearing an incidence of 52.9%, which indicated that the occurrence of hypophosphatemia was associated with patients' age, primary disease and postoperative intravenous hypertonic glucose solution transfusion. Meanwhile, the existance of hypophosphatemia was often complicated by hypocalcemia (83.1%), hyponatremia (76.4%), and hypokelemia (37.5%), and furthermore, was in connection with organ dysfunction or failure. Of the 40 death cases among all 257, 31 endured hypophosphatemia (77.5%). Hence, prompt diagnosis and active prevention and treatment of hypophosphatemia are essential to raise the cure rate of surgical critical patients.

Adolescent↗

[Free radical production in the subdermal vascular network thin skin flap].

In this experiment 12 piglets were used to study the survival length and free radical production of subdermal vascular network thin skin flap, fascial flap and PSVN. The flap or graft was designed 15 cm x 3 cm in size on the flanks of the animals. Twenty-four flaps or grafts were formed in each group. The observation on 7 postoperative day indicated that the survival length of the subdermal vascular network thin skin flap was 38% over that of fascial flap, while the free radical was less in the former, and it could be one of the reasons for its better survival.

Animals↗

Relationship of LDLR gene polymorphism and NIDDM in Chinese.

Genomic DNA was extracted from peripheral blood lymphocytes of 105 healthy and 75 NIDDM Chinese subjects. The fragment located in exon 13 of the low density lipoprotein receptor (LDLR) gene was amplified by polymerase chain reaction (PCR), and digested with restriction enzyme HincII. LDL, TC and TG levels were measured in all subjects. Investigations were conducted to explore the correlation between the HincII RFLP of LDLR gene and NIDDM in the Chinese population. The results showed that no significant correlation existed between this RFLP locus and NIDDM. Marked differences were found, however, between the genotype distribution of low LDL level subgroups of NIDDM patients and normal controls. It was inferred that the H1 allele might be associated with high blood cholesterol levels, and the H2 allele with low cholesterol levels. Disturbances of lipid metabolism occur frequently in diabetes mellitus. This study suggested that differences in LDLR genotypes may affect the phenotypes of lipid metabolism.

Adult↗

The population association of glucokinase gene with type 2 (noninsulin-dependent) diabetes mellitus in Chinese.

The association of gluckinase (GCK) gene with type 2 (non-insulin-dependent) diabetes mellitus was investigated in 168 Chinese subjects (85 unrelated type 2 diabetics and 83 non-diabetic controls). The microsatellite polymorphism marker, GCK-5', was amplified with polymerase chain reaction. Four alleles were observed in Chinese population with length varying from 137bp to 143bp and the most common one being the 139bp allele 3. In comparison with non-diabetics, allele 4 was significantly increased in type 2 diabetes (10% versus 38, respectively; X2 = 6.773, P = 0.009); genotype 44 and 4X (X denotes any allele other than allele 4) were significantly increased in type 2 diabetes (16% versus 6% respectively; X2 = 6.439, P = 0.011). The frequency difference was also shown in overweight/obese subgroup comparison (X2 = 7.718, P = 0.021), but not in lean/normal-weight subgroup comparison. No differences of age of onset and frequency of positive family history were observed between type 2 diabetic patients with genotype 44 or 4X and those with XX. The risk for type 2 diabetes in Chinese with genotype 44 or 4X was about 3.5 times higher than in Chinese with genotype XX. Therefore, GCK gene was associated with Chinese type 2 diabetes.

Adult↗

Lung cancer, smoking patterns, and mutagen sensitivity in Mexican-Americans.

BACKGROUND: Mexican-Americans have lower age-adjusted lung cancer incidence rates than non-Hispanic whites and African-Americans. Since 87% of lung cancers are attributed to tobacco exposure, this difference could be explained partly by lower prevalence of cigarette smoking. However, only a fraction of exposed individuals will develop smoking-related cancer, and genetically determined differences in modulation of environmental exposures could also explain some of this ethnic risk differential in lung cancer incidence in the United States. However, little research on genetic susceptibility has been focused on Hispanic populations in the United States. METHODS: We are conducting a case-control study of lung cancer in a high-risk group (African-Americans) and a low-risk group (Mexican-Americans) to evaluate ethnic differences in mutagen sensitivity by an in vitro assay that quantifies mutagen-induced chromosome breaks in short-term lymphocyte cultures. RESULTS: In the 174 Mexican-Americans (67 lung cancer case patients and 107 control subjects) accrued to date, all measures of cigarette smoking (intensity, duration, nicotine and tar contents, depth of inhalation, and type of cigarette) were significant predictors of lung cancer risk. There were significantly higher risks associated with mutagen sensitivity (defined as > or = 1 break/cell) for both former smokers (odds ratio [OR] = 4.5; 95% confidence interval [CI] = 0.9-21.9) and current smokers (OR = 2.6; 95% CI = 0.6-11.1). Mutagen sensitivity also appeared to be implicated in risk in patients who were less than 55 years old at diagnosis (OR = 15.0; 95% CI = 1.0-228.9) and in those with lower cigarette exposure (OR = 11.0; compared with an OR of 1.7 for the heaviest smokers). The overall OR for mutagen sensitivity adjusted for age, sex, and pack-years of smoking was 2.9 (95% CI = 0.8-9.9). Neither current smoking status nor years of exposure shifted the sensitivity profile of case patients and control subjects. CONCLUSION: Although this study showed higher percentages of nonsmokers among Mexican-Americans than our previously reported data for African-Americans, the Mexican-American case patients were heavier smokers than the African-American case patients. The prevalence of mutagen sensitivity for Mexican-Americans was 64.1% in case patients and 26.2% in control subjects. In African-Americans, mutagen sensitivity was previously reported to be 55.3% in case patients and 24.6% in control subjects. These preliminary data do not support our a priori hypothesis that a lower prevalence of mutagen sensitivity in Mexican-Americans would account for the lower incidence of lung cancer. Mutagen sensitivity, however, is only one of an array of potential susceptibility markers that we are evaluating in this unique population.

Black or African American↗

[Karyotype analysis of Candida species by PFGE and its use in classicification].

Electrophoretic karyotype in 7 medically important Candida species was analyzed by pulsed-field gel electrophoresis. The chromosomal DNAs of the Candida species were separated into 4-10 bamds ranging in size from 0.5-2.8 Mb. These patterns were species-specific, so the electrophoretic karyotype can be used as a genotype basis for classification and identification of Candida species.

Candida↗

[Study on the perinatal infection caused by Chlamydia trachomatis].

OBJECTIVE: Study on the frequency and outcome of chlamydia trachomatis (CT) infection of the pregnant women and their babies. METHODS: Two hundred and seventy eight pregnant women and their 79 infants were studied with McCoy cell culture, polymerase chain reaction (PCR) and DNA sequence analysis. Isolation of CT was performed in McCoy cell culture, and iodine-stained glycogen-positive inclusions were considered to be positive. CT DNAs were extracted by modified NaI method. Two sets of primers derived from CT common plasmid and major outer membrane protein genes were used in the PCR. Dideoxy-mediated chain-termination method was used for DNA sequence analysis. RESULTS: 10.8% (30/278) of the cervical cultures were positive for CT, while the positive rate by PCR was 14.0% (39/278). The vertical transmission rate of CT was 55.0% (11/22). The sequences of DNA amplified from a maternal-infant pair were matched perfectly. The incidence of puerperal endometritis in CT positive women was 28.6%, much higher than 8.1% in those CT negative mothers. Neonatal conjunctivitis and pneumonia in group of CT positive mothers were 45.0% and 30.0%, respectively, much higher than 18.3% and 8.3%, respectively, in that of CT negative mothers (P < 0.05). CONCLUSION: Perinatal infection caused by CT is very common in China.

Chlamydia Infections↗

[The clonal origin of multiple myeloma].

In order to explore the clonal origin and malignant pathway of multiple myeloma (MM), a study has been made to investigate the differentiation stage of malignant progenitor cells in peripheral blood (PB) of MM. The immnophenotype of mononuclear cells (MC) in PB and bone marrow (BM) from 17 patients with MM were studied with APAAP method by using monoclonal antibodies directed to a series of B-cell markers (CD10, CD19, CD20, CD45RA, CD45RO, CD38). There were no plasma cells in samples of PB examined. Out of the 17 patients, MC of PB from 10 expressed plasma cell antigen CD38. This expression was significantly different from normal control (P < 0.001) and identical with that of BM. MC of 9 patients were CD38+ and CD45RO+, 3 were CD38+, CD45RO+ and CD45RA+ and 1 was CD38+, CD45RA+, CD45RO+ and CD10+ in PB. The results indicate that while the B-cell marker's expression of PB matches with that of BM, its forms are diversified. This suggests that the PB of MM patients contains precursors at multiple differentiation stages of myeloma cells, from early B cell to active B cell, late B cell and preplasma cell. MC of PB from 10 patients expressed CD45RO while that of BM from 5 out of them expressed the same. This indicates that preplasma cells which are CD45RO+ in PB may eventually return to BM for homing and differentiating into plasma cells, thus losing CD45RO and expressing CD38 only. This study has proved the presence of malignant cells in PB of MM, confirmed the speculation of the malignant pattern and provided the theoretical basis for purging of PB in autologous peripheral blood stem cell transplantation.

Adult↗

Regulation of an eukaryotic initiation factor-2 (eIF-2) associated 67 kDa glycoprotein (p67) and its requirement in protein synthesis.

The p67 mRNA level and p67 requirement in protein synthesis were studied using an animal cell (KRC-7, rat tumor hepatoma cell) in culture. p67 mRNA was present in confluent cells but disappeared almost completely from serum-starved cells. However, when PMA was added to the serum-starved cells, p67 mRNA appeared in increasing quantities. Several-fold molar excess of p67 mRNA over that present in confluent cells was detected within 2 h of PMA addition and this level remained the same during the 4 h of the experiment. p67 requirement in protein synthesis was studied using a p67 antisense DNA construct under a metallothionein gene promoter. Expression of this antisense DNA in the presence of zinc in PMA-induced serum-starved cells completely inhibited induced appearance of p67 mRNA and subsequent protein synthesis. These results suggest that p67 is regulated at the mRNA level and also that this protein factor is essential for protein synthesis.

Aminopeptidases↗

Effect of taxol and diamide on shear tolerance of hybridoma and insect cells.

The role of the cytoskeletal structure on the shear tolerance of hybridoma and insect cells to laminar and turbulent shear flow field was investigated. The cytoskeletal network of cells modified by taxol and diamide, which are known to alter the stability of specific cytoskeletal components, resulted in increased cell shear tolerance to both laminar and turbulent flow fields. Plasma membrane fluidity measurements revealed that modification of microtubules by taxol did not alter the fluidity state of the plasma membrane. The increase in shear tolerance by taxol treatment is attributed to its ability to stabilize microtubules in the cytoskeleton. A low concentration of taxol, 0.05 microM, did not reduce the hybridoma cell growth rate or the monoclonal antibody production rate, but both were decreased at a higher concentration, 0.25 microM.

Animals↗

The tyrosine kinase activity of the C-erbB-2 gene product (p185) is required for growth inhibition by anti-p185 antibodies but not for the cytotoxicity of an anti-p185-ricin-A chain immunotoxin.

Our previous studies have demonstrated that 7 of 10 IgG antibodies against distinct epitopes on the extracellular domain of the c-erbB-2 gene product (p185) inhibit the anchorage-independent growth of SKBr3 human breast-cancer cells that overexpress this transmembrane tyrosine kinase growth-factor receptor. Two of 7 growth-inhibitory antibodies also block the binding and function of the gp30 and p75 c-erbB-2 ligands. In this report we have studied phosphorylation of p185 and different intracellular substrates after binding of antibodies that do or do not inhibit tumor-cell growth. A correlation has been found between antibodies that inhibit growth and the intensity of tyrosine phosphorylation of p185. At late intervals, serine phosphorylation of at least 3 intracellular substrates is increased preferentially by growth-inhibitory antibodies. To test the importance of p185 kinase activity more critically, NIH3T3 cells were transfected with an expression vector containing the full-length human c-erbB-2 gene (cell line 17313), c-erbB-2 with deletion of the kinase region from codons 751-979 (cell line 9309) or c-erbB-2 with deletion of most of the intracellular domain from codons 684-1255 (cell line 9310). Unconjugated antibodies inhibited anchorage-independent growth of 17313 cells as well as SKBr3 cells, but did not inhibit growth of either 9309 or 9310 cells. In contrast, the cytotoxic effect of anti-p185-ricin A chain (RTA) conjugates was comparable for 17313, 9309 and 9310. The tyrosine-kinase activity of p185 is required for growth inhibition mediated by unconjugated anti-p185 antibodies, but not for the cytotoxic activity of anti-p185-RTA immunotoxins.

3T3 Cells↗

Retinoid-resistant estrogen receptor-negative human breast carcinoma cells transfected with retinoic acid receptor-alpha acquire sensitivity to growth inhibition by retinoids.

Retinoids mediate their actions via RARs (retinoic acid receptors) and RXRs (retinoid X receptors). Each class of these nuclear retinoid receptors is further subdivided into three species, namely alpha, beta, and gamma. Recent studies demonstrate that estrogen receptor (ER)-positive human breast carcinoma (HBC) cell lines and tumor samples exhibit significantly higher levels of RAR alpha than their ER-negative counterparts. ER-positive HBC cell lines are sensitive to, and ER-negative cell lines are resistant to, growth inhibitory effects of retinoic acid (RA). We previously demonstrated that the expression of functional ERs in an established ER-negative cell line resulted in higher levels of RAR alpha and sensitivity to growth inhibition by RA. To further investigate the major role of RAR alpha in retinoid-mediated inhibition of growth, we transfected RAR alpha cDNA in two RA-resistant ER-negative HBC cell lines. Analyses of different clonal populations of RAR alpha transfectants from each cell line revealed growth inhibition by retinoids. Utilizing RAR- and RXR-class selective retinoids, we further demonstrated that only the RAR alpha-selective retinoids mediated the growth inhibition in these cells, while the RXR-selective retinoids were biologically inert. We thus provide evidence that the molecular mechanisms of retinoid inhibition of HBC proliferation predominantly involve RAR alpha.

Base Sequence↗

Iron protoporphyrin IX (hemin) but not tin or zinc protoporphyrin IX can stimulate gene expression in K562 cells from enhancer elements containing binding sites for NF-E2.

Many genes whose transcription is erythroid-specific contain enhancer or promoter elements that bind the transcription factor NF-E2. Hemin induction increases the expression of globin genes in the human erythroleukemia cell line K562, and increases the expression of reporters gene regulated by an enhancer elements containing binding sites for NF-E2. The failure of metalloporphyrins other than hemin to stimulate the transient expression of a CAT reporter gene linked to an enhancer element containing a binding site for NF-E2 was correlated with their failure to induce benzidine-positive K562 cells and increase the steady-state level of gamma-globin mRNA. This study suggests that elevated levels of zinc protoporphyrin IX found in the anemia of chronic disease, iron deficiency, and lead poisoning may contribute to a decrease in globin gene expression by interfering with the transcriptional activity of enhancer elements containing binding sites for NF-E2.

Base Sequence↗

A plasma lipoprotein containing only apolipoprotein E and with gamma mobility on electrophoresis releases cholesterol from cells.

Previous studies have identified lipid-poor high density lipoproteins with electrophoretic pre-beta mobility as the initial acceptors of cell-derived cholesterol in human plasma. These lipoproteins contain apolipoprotein A-I (apo A-I) as their sole apolipoprotein. In the present study, incubation of human plasma with [3H]cholesterol-laden skin fibroblasts has led to the identification of another lipoprotein that serves as a potent initial acceptor of cell-derived cholesterol. This lipoprotein, which we term gamma-LpE, exhibits gamma mobility on agarose gel electrophoresis. As determined by nondenaturing PAGE and by electron microscopy, the size of the spherical particle ranges between 12 and 16 nm. SDS/PAGE and subsequent immunoblotting identified apoE as its sole apolipoprotein. Plasma from normal and apoA-I-deficient mice, but not from apoE-deficient mice, released [3H]cholesterol from fibroblasts into a gamma-migrating lipoprotein. Cell culture media from hepatoma cells or mouse peritoneal macrophages, both of which contain apoE of cellular origin, also promoted efflux of [3H]cholesterol from fibroblasts into a gamma-migrating fraction. This was not observed with cell culture medium from fibroblasts alone. In conclusion, our results strongly indicate the presence in human plasma of a lipoprotein containing only apoE, gamma-LpE, which is secreted by peripheral cells and is a potent acceptor of cell-derived cholesterol.

Animals↗

Detection of herpes simplex viral DNA in the iridocorneal endothelial syndrome.

OBJECTIVE: To test the hypothesis that the iridocorneal endothelial (ICE) syndrome has a viral origin by comparing the incidence of viral DNA in corneal specimens from patients with the ICE syndrome and from controls. DESIGN: Thirty-one corneas obtained from 25 patients with the ICE syndrome and six with chronic herpetic keratitis (n = 31) were compared with 30 control specimens obtained from 15 healthy donors and from 15 patients with other, nonviral chronic corneal diseases. METHODS: Primer pairs and polymerase chain reaction methods were used to identify and amplify either a segment of the DNA polymerase gene in the case of the herpes simplex and zoster viruses or a region of the nuclear antigen gene for the Epstein-Barr virus. The oligonucleotide amplified by polymerase chain reaction was fully characterized with the use of restriction enzyme, hybridization, and sequence analyses to determine that it contained the expected base pair sequence. RESULTS: Sixteen of 25 ICE syndrome specimens and four of six herpetic keratitis specimens were positive for herpes simplex virus (HSV) DNA. All nine ICE syndrome specimens tested were negative for the presence of DNA from the herpes zoster or the Epstein-Barr viruses. Controls were uniformly negative for HSV DNA whether they were obtained from ostensibly normal corneas (n = 15) or from corneas with intestinal keratitis, aphakic bullous keratopathy, or keratoconus (n = 15). Tissue samples cut from positive ICE syndrome specimens yielded negative results when retested after the endothelial layer was removed. These findings indicate that localization of HSV DNA is within the endothelium, the tissue primarily involved in the pathogenesis of the ICE syndrome. CONCLUSIONS: Polymerase chain reaction evidence shows that HSV DNA is present in a substantial percentage of ICE syndrome corneal specimens and that HSV-DNA is absent in normal corneas and in corneas from patients with three other chronic corneal diseases. These results provide direct evidence to support our hypothesis that the ICE syndrome has a viral origin. We discussed clinical implications, including possible therapeutic interventions.

Base Sequence↗