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Biomedical subjects

Y Fu

Publications and source records attributed to Y Fu.

At least 163 records · Page 9Linked to original sources

[Circulating ICAM-1 in sera of renal allograft recipients in monitoring of acute rejection].

OBJECTIVE: To study the significance of ICAM-1 in sera of renal allograft recipients in mornitoring of acute rejection. METHODS: The levels of circulating intercellular adhesion molecule-1 (cICAM-1) in sera of 87 renal transplantation recipients, including 29 patients with acute rejection and 8 patients with cyclosprine nephrotoxicity were measured by sandwich-ELISA. RESULTS: The levels of cICAM-1 were elevated transiently after transplantation. They decreased and reached normal levels within 2 weeks. In the acute rejection episode, the levels of cICAM-1 were significantly higher than those of patients with stable graft function and cyclosporine nephrotoxicity (P < 0.001). The increase of cICAM-1 was seen 1-3 days early than the diagnosis was made. On successful antirejection therapy, the levels of cICAM-1 decreased to normal, but in patients with steroid-resistant rejection, it did not decrease. CONCLUSION: The cICAM-1 in sera can be used to estimate the graft function, and it is also useful in prediction and differential diagnosis of acute rejection and prognosis.

Adult↗

[The pressure of amniotic cavity during pregnancy and treatment of unruptured oligohydramnios with antepartum amnioinfusion].

OBJECTIVES: To measure the pressure of the amniotic cavity during pregnancy and to study the effects of antepartum amnioinfusion on oligohydramnios with intact membranes. METHOD: We used an improved three-way switch apparatus to measure 110 women during pregnancy, and treated 97 oligohydramnios with antepartum amnioinfusion or intravenous infusion as controls. RESULTS: The pressure of the amniotic cavity was 1.69 +/- 0.23 kPa, which was close to the normal value in pregnancy. In oligohydramnios, it was 1.1 +/- 0.3 kPa, and in polyhydramnios 3.2 +/- 0.3 kPa(P < 0.01-0.001). When 300 ml fluid was amnioinfused, the amniotic fluid index (AFI) increased by 5.0 +/- 1.8 cm, the pressure of amniotic cavity increased by 0.18 +/- 0.2/kPa and the incidence of vaginal delivery was 87.9%, whereas that of the control group was 12.1%. There were no fetal distress, asphyxia neonatorum, amnionitis or ruptured membrane. CONCLUSIONS: The pressure of the amniotic cavity during normal pregnancy was stable, but changed with AFI. Antepartum amnioinfusion for unruptured oligohydramnios resulted in a marked increase in incidence of vaginal delivery, a decreased ratio of cesarean section and a reduction of perinatal morbidity. These differences are highly significant.

Adult↗

[Microsurgical technique in the treatment of lumbar disc prolapse].

From 1983 to 1996, 354 cases of the lumbar intervertebral disc prolapse were treated by the microsurgical technique in our hospital. Among them, 330 cases were the simple prolapse of the lumbar intervertebral disc, the other 24 cases were complicated with the lateral recess stenosis. The application of the microsurgical technique has the advantage of its precision, shorter operative time, little surgical trauma, less bleeding, and quicker recovery. So the patients can usually get up and move around at an earlier time. Ninety-five percent of the patients acquired good results. It is suggested that the microsurgical technique is a relatively good operative method in the treatment of the lumbar intervertebral disc prolapse and the lateral recess stenosis.

Adolescent↗

[Rapid determination of essential fatty acids of edible oils by conversion to their hydroxamic acids].

A simple and rapid HPLC method for the determination of essential fatty acids of edible oils was established. Oil samples were converted to their hydroxamic acids in a single step and analyzed without prior separation and purification. The chromatographic conditions were: Shim-pack CLC ODS, 5 microns, 150 mm x 6.0 mm i.d. column, 40 degrees C; MeOH: 0.02 mol/L NaH2PO4(pH 3.0) (81:19, V/V) as eluent and UV-213 nm detector. The linear range was 0.05-0.6 g/L, recovery was 96.93% and RSD was 1.80%(n = 4). The relative standard deviations for intra-day and inter-day assays were 1.24% and 1.62% respectively (n = 6). For 18:3, 18:2, 18:1, the difference between the derivatization yields from triglycerides and their methyl esters was almost one fold. That was confirmed by our recovery and determination results. The calibration curves for the oil samples should not be obtained from the derivatization of their methyl ester standards.

Chromatography, High Pressure Liquid↗

[Living related liver transplantation: a case report].

OBJECTIVE: To study a case of living related liver transplantation. METHOD: The patient was a 10-year-old girl who suffered from congenital diffuse intrahepatic cholangiectasis, recurrent cholangitis and hepatocirrhosis. The donor was the patient's father aged 40. The left lateral lobe of donor's liver was cut and grafted to the patient. Intensive care and treatment as well as follow-up were given after operation. RESULT: For the donor, the operation lasted 400 min, with 410 ml bleeding and 300 g liver removed. The recovery was satisfactory. For the recipient, the operation lasted 652 min, with 1665 ml bleeding, 80 min non-liver stage, 0 min graft hot ischemic stage, and 137 min cold ischemic stage. Immunosuppressive therapy was given using cyclosporin A, azathioprine and adrenocortical hormones. There was an acute rejection 11 days after operation, which was controlled using hormone impulsive therapy. The patient has been surviving 7 months with normal liver function. CONCLUSION: The living related liver transplantation is feasible under modern surgical conditions. It is demonstrated that perfect postoperative management is the key for the successful liver transplantation.

Adult↗

[Expression of nm23 in gastrointestinal smooth muscle tumors and its relation to cell proliferative activity].

OBJECTIVE: To determine the relationship between nm23 expression and benign or malignant degree, metastasis, prognosis and cell proliferative activity of gastrointestinal smooth muscle tumors (GISMT). METHOD: 86 cases of GISMT were studied, and nm23 was detected by the immunohistochemical staining S-P method. The cell proliferative activity was evaluated by the silver colloid method for argyrophilic nucleolar organizer regions proteins (AgNORs) and by the immunohistochemical staining S-P method for proliferating cell nuclear antigen (PCNA). RESULT: The expression of nm23 declined significantly according to the following order: leiomyomas, low malignant leiomyosarcomas, high malignant leiomyosarcomas (P < 0.01). The expression of nm23 was associated with the tumors with or without contiguous organ invasion or distant metastasis, the size of tumors, the tumors with or without center necrosis (P < 0.01 or P < 0.05). The five-year survival rate was significantly higher in patients with nm23 positive expression than that with nm23 negative expression (P < 0.05). The expression of AgNORs and PCNA with nm23 negative cases was obviously higher than that with nm23 positive cases (P < 0.01). CONCLUSION: nm23 is a valuable indicator for the biological characteristics of GISMT. nm23 expression and cell proliferative activity can supply a deficiency each other in distinguishing malignant from benign tumors, judging the malignant degree, and predicting the prognosis of the patients with GISMT.

Adolescent↗

[Studies on flavones of Epimedium bevicorum Maxim].

OBJECTIVE: To systematically develop Epimedium spp. and control their quality. METHOD: Using polyamide and macroporous adsorbent resins AB-8. The compounds were identified on the basis of spectrometric data and physical and chemical test. RESULT: Four flavones were isolated from the herb. They were identified as wushanicariin, baohuoside VI, kaempferol-3,7-O-alpha-L-dirhamnoside and hexandraside E. CONCLUSION: These flavones were isolated from the herb and identified for the first time.

Drugs, Chinese Herbal↗

Nicotine-induced norepinephrine release in the rat amygdala and hippocampus is mediated through brainstem nicotinic cholinergic receptors.

Previous studies have shown that nicotine stimulates norepinephrine (NE) release in the rat hypothalamic paraventricular nucleus, which in turn activates the hypothalamo-pituitary-adrenal axis. In the present study, nicotine induced NE release in the amygdala (AMYG) and the hippocampus (HP) of the same rat in vivo. Nicotine (0.065-0.135 mg/kg i.v. at a rate of 0.09 mg/kg/60 sec) dose-dependently increased NE release at both sites with similar potencies. To determine whether the site of action of nicotine is in the brainstem, which contains the noradrenergic cell bodies projecting to AMYG and HP, nicotinic cholinergic receptor (NAchR) antagonists were injected into the cerebral aqueduct before i.v. nicotine. Use of the following antagonists enabled partial characterization of the NAchRs mediating NE secretion: mecamylamine (Mec), dihydro-beta-erythroidine (DH beta E), methyllycaconitine (MLA) and alpha-bungarotoxin (alpha-BTX). Mec inhibited 80% of NE release in AMYG and 87% in HP (IC50 = 6 nmol for both regions). DH beta E blocked 62% of NE release in AMYG (IC50 = 8 nmol) and 63% in HP (IC50 = 15 nmol). Similar to DH beta E, MLA inhibited 60% of NE release in AMYG and 66% in HP (IC50 = 5 nmol for both regions). In contrast, alpha-BTX had no effect on NE release in either region. These results indicate that brainstem NAchRs accessible from the fourth ventricle mediate nicotine-stimulated NE secretion in AMYG and HP. Taken together with prior investigations showing the brainstem expression of mRNAs encoding NAchR subtypes and the selectivity of antagonists for NAchR subtypes, the present studies suggest that brainstem alpha-3 subunits may be involved.

Amygdala↗

[The types and distributive features of gene mutation on beta-thalassemia in three districts in northwest China].

OBJECTIVE: To sum up the results of gene analyses on beta-Thalassemia in three districts in northwest China and discuss the types and distributive features of the mutation. METHODS: Polymerase china reaction in combination with dot-blot hybridization of allele-specific oligonucleutide probes(PCR-ASO). RESULTS: In the gene analysis carried out in 85 probands with beta-Thalassemia, twelve types of gene mutation were identified from five nationalities. CD8(-AA), CDs8-9(+G), CDs27-28(+C) heterozygote and [-28(A-->uG). CD17(A-->uT)/N] double heterozygote were firstly assayed in these districts besides the common types in Chinese, of which, [-28(A-->uG). CD17(A-->uT)/N] as double gene mutation in the same chromosome is rare. CONCLUSION: There are significant differences in the distribution and nationality features of gene mutation types for beta-Thalassemia in the three districts, so the three districts as a whole is an area with special distributive features of the disease.

Adolescent↗

Characterization and chromosomal localization of PTPRO, a novel receptor protein tyrosine phosphatase, expressed in hematopoietic stem cells.

Hematopoietic stem cells (HSCs) support blood cells throughout life by utilizing their self-renewing and multilineage differentiating capabilities. Hematopoietic growth factors mediate their effects on stem cells by the tyrosine phosphorylation of proteins. Regulation of tyrosine phosphorylation is partially mediated by protein tyrosine phosphatases (PTPases). A possible mechanism by which hematopoietic stem cells maintain their self-renewing capacity and undifferentiated state is by controlling the balanced and opposing actions of protein tyrosine kinases (PTKs), receptors for growth factors, and PTPases. We have characterized the expression of PTPases in 5-fluorouracil (5-FU)-treated murine bone marrow cells, which represent a very primitive population of progenitors enriched for reconstituting stem cells, by using a consensus polymerase chain reaction (PCR) method. Several PTPases were expressed abundantly in the 5-FU-treated bone marrow stem cells. A novel PTP, termed protein tyrosine phosphatase receptor omicron (PTPRO), which is related to the homotypically adhering kappa, mu and PCP-2 receptor-type tyrosine phosphatases, was identified and characterized. We have cloned the murine and full-length human PTPRO cDNAs which share 89% homology, indicating that PTPRO is highly conserved between these species. The human PTPRO cDNA clone encodes a polypeptide of 1439 amino acids (aa) and has a calculated molecular mass of approximately 162 kDa. PTPRO consists of an extracellular segment containing a MAM domain, an immunoglobulin (Ig) domain, four fibronectin-type III (FN-III) repeats, a transmembrane segment, and two tandem intracellular PTP domains. The human PTPRO gene was assigned to human chromosome 1p35-pter using Southern blot analyses of genomic DNAs from rodent/human somatic hybrid cell lines containing human chromosome 1 or the p35-pter region of the chromosome. The mouse Ptpro gene was mapped to chromosome 4, closely linked to D4Mit16 and Elp1 (elliptocytosis-1), by using genomic DNAs from a (C57BL/6J x Mus spretus)F1 x Mus spretus backcross. In fetal tissues, PTPRO expression was observed in the brain and lung, whereas lower levels were observed in the kidney. In adult tissues, PTPRO was less restricted and was observed in the lung, heart, skeletal muscle, prostate, testis, and in various areas of the brain, indicating that PTPRO expression is developmentally regulated. Expression of PTPRO was also observed in human CD34+ bone marrow cells and 5-FU-treated murine primitive stem cells. These results suggest a potential role for PTPRO in stem cell adhesion and in mediating homophilic cell-cell interactions in other cell types.

Amino Acid Sequence↗

Isolation from phage display libraries of single chain variable fragment antibodies that recognize conformational epitopes in the malaria vaccine candidate, apical membrane antigen-1.

Phage display of single chain variable fragment (scFv) antibodies is a powerful tool for the selection of important and useful antibody specificities. We have constructed such a library from mice protected from malaria challenge by immunization with recombinant Plasmodium chabaudi DS apical membrane antigen (AMA-1). Panning on refolded AMA-1 enriched a population of scFvs which specifically bound the antigen. The single chain antibodies recognize conformational epitopes on AMA-1 from the P. chabaudi DS strain but not on AMA-1 of the 556KA strain of P. chabaudi. A subset of the antibody fragments recognized AMA-1 from the human malaria parasite Plasmodium falciparum. Nucleotide sequencing revealed that at least four unique scFv genes were selected by the panning procedure. These scFv antibodies are valuable reagents for probing the structure and function of AMA-1 and will be used to test the feasibility of using recombinant antibodies in a passive immunization therapy against malaria.

Animals↗

Fib420, the novel fibrinogen subclass: newborn levels are higher than adult.

Fib420 is a recently identified subclass of normal human fibrinogen in which two extended alpha chain isoforms (alphaE) replace the common alpha chains, yielding a molecule (ca. 420 kD) which is larger than the more abundant 340-kD form. Evidence for preservation of this subclass throughout vertebrate evolution suggests it performs some as yet unidentified vital function. A survey was undertaken to establish the range of plasma Fib420 levels in normal, healthy adults and in placental cord (fetal) blood. For measuring Fib420, a quantitative Western blot assay was developed using monoclonal antibody against the exon-VI encoded C-terminus of the molecule's unique alphaE chain. This alphaE chain signal was normalized to that of the beta chain, common to both fibrinogen forms. Analysis of plasma samples from the adult and newborn cohorts (n = 25 each; total fibrinogen ca. 2.6 mg/mL in both) revealed a statistically significant difference, with a mean level of 100 +/- 28 microg/mL in the neonate compared to 34 +/- 7 microg/mL in the adult. On average, 1 out of every 100 fibrinogen molecules in adult plasma belongs to the Fib420 subclass. Unlike in the newborn, adult Fib420 levels remained the same over a wide range of total plasma fibrinogen. The striking difference observed between these two cohorts suggests a changing developmental expression of the Fib420 subclass and a homeostatic control operating in later stages of life.

Adult↗

Reduced expression of Tap1 and Lmp2 antigen-processing genes in the nonobese diabetic (NOD) mouse due to a mutation in their shared bidirectional promoter.

The MHC is an essential contributor to autoimmunity. Lmp2 and Tap1 are genes located in the MHC class II region, and they encode proteins participating in the generation and transport of endogenous peptides for T cell education. A mutation (T-->A) has now been detected in the shared bidirectional promoter of the Lmp2 and Tap1 genes in the nonobese diabetic (NOD) mouse. The nucleotide substitution (TCATTC-->TCAATC) in NOD mice eliminates an initiator (Inr) element (TCATTC) thought to be important for RNA polymerase II positioning in the Lmp2 orientation. It also created a CAAT-like box and an inverted CAAT-like box in the Lmp2 and Tap1 orientations, respectively. Northern blot revealed reduced amounts of Tap1 and Lmp2 mRNA in NOD mice, and 5'-rapid amplification of cDNA ends revealed the loss of a transcription start site of Lmp2 in these animals. The Tap1-Lmp2 promoter from NOD mice showed reduced transcriptional activity in transient transfection assays with luciferase reporter constructs for both Tap1 and Lmp2 genes. Observed altered substrate specificity of Lmp2 containing proteasomes isolated from NOD mice was consistent with reduced Lmp2 activity. The beneficial influence of non-MHC genes (NOR mice) and gender factors (male NOD mice) influencing the penetrance of the promoter polymorphism further confirmed the essential gender and hormonal context of the mutation. This study identifies the first specific mutation in the MHC of the NOD mouse that specifically impacts the activity of genes involved in peptide presentation, a process essential for T cell education.

ATP Binding Cassette Transporter, Subfamily B, Mem↗

The related adhesion focal tyrosine kinase differentially phosphorylates p130Cas and the Cas-like protein, p105HEF1.

The related adhesion focal tyrosine kinase (RAFTK) is tyrosine-phosphorylated following beta1 integrin or B cell antigen receptor stimulation in human B cells. Two substrates that are tyrosine-phosphorylated following integrin ligation in B cells are p130(Cas) and the Cas family member human enhancer of filamentation 1 (HEF1), both of which can associate with RAFTK. In this report we observed that RAFTK was involved in the phosphorylation of these two proteins. While a catalytically active RAFTK was required for both p130(Cas) and HEF1, phosphorylation of p130(Cas), but not of HEF1, was dependent on an intact autophosphorylation site (Tyr402) on RAFTK. To determine if RAFTK phosphorylated p130(Cas) and HEF1 directly or through an intermediate, we assayed the ability of RAFTK and of a Tyr402 mutant to phosphorylate purified HEF1 and p130(Cas) domains. RAFTK was able to phosphorylate the substrate domains of both p130(Cas) and HEF1, but only the C-terminal domain of p130(Cas). Furthermore, Tyr402, which mediates the binding of RAFTK to c-Src kinase, was required for the phosphorylation of the C-terminal domain of p130(Cas). These data suggest that RAFTK itself is sufficient for HEF1 phosphorylation, whereas a cooperation between RAFTK and Src kinases is required for the complete phosphorylation of p130(Cas).

Adaptor Proteins, Signal Transducing↗

Tyrosine phosphorylation of the related adhesion focal tyrosine kinase in megakaryocytes upon stem cell factor and phorbol myristate acetate stimulation and its association with paxillin.

We have characterized signaling pathways involving the related adhesion focal tyrosine kinase (RAFTK, also known as PYK2 or CAK-beta) in CMK human megakaryocytic cells. Stem cell factor, which potentiates the growth of megakaryocytes and their progenitors, and phorbol myristate acetate, which causes differentiation of megakaryocytic cell lines, induced the tyrosine phosphorylation of RAFTK but not of focal adhesion kinase. Stimulation of CMK cells with stem cell factor resulted in an increase in the autophosphorylation and kinase activity of RAFTK. Phosphorylation of RAFTK under these conditions was mediated by a protein kinase C-dependent pathway. Cytochalasin D, which disrupts the cytoskeleton, abolished the phosphorylation of RAFTK upon phorbol myristate acetate and stem cell factor stimulation, indicating that RAFTK association with the actin cytoskeleton appears to be critical for its phosphorylation. In addition, we observed an association of RAFTK with paxillin, a 68-kDa cytoskeleton protein. Using in vitro binding assays, RAFTK and paxillin were shown to bind directly through the C-terminal proline-rich domain. Transient overexpression of a dominant-negative mutant of RAFTK inhibited significantly the tyrosine phosphorylation of paxillin upon phorbol myristate acetate stimulation. These observations indicate that RAFTK might play an important role in the phosphorylation of signaling pathways within the focal adhesions and that RAFTK participates in signaling events that link signals from the cell surface to the cytoskeleton. Furthermore, this study suggests that RAFTK might be involved in megakaryocyte proliferation and differentiation.

Binding Sites↗

Direct association of Csk homologous kinase (CHK) with the diphosphorylated site Tyr568/570 of the activated c-KIT in megakaryocytes.

The Csk homologous kinase (CHK), formerly MATK, has previously been shown to bind to activated c-KIT. In this report, we characterize the binding of SH2(CHK) to specific phosphotyrosine sites on the c-KIT protein sequence. Phosphopeptide inhibition of the in vitro interaction of SH2(CHK)-glutathione S-transferase fusion protein/c-KIT from SCF/KL-treated Mo7e megakaryocytic cells indicated that two sites on c-KIT were able to bind SH2(CHK). These sites were the Tyr568/570 diphosphorylated sequence and the monophosphorylated Tyr721 sequence. To confirm this, we precipitated native CHK from cellular extracts using phosphorylated peptides linked to Affi-Gel 15. In addition, purified SH2(CHK)-glutathione S-transferase fusion protein was precipitated with the same peptide beads. All of the peptide bead-binding studies were consistent with the direct binding of SH2(CHK) to phosphorylated Tyr568/570 and Tyr721 sites. Binding of FYN and SHC to the diphosphorylated Tyr568/570 site was observed, while binding of Csk to this site was not observed. The SH2(CHK) binding to the two sites is direct and not through phosphorylated intermediates such as FYN or SHC. Site-directed mutagenesis of the full-length c-KIT cDNA followed by transient transfection indicated that only the Tyr568/570, and not the Tyr721, is able to bind SH2(CHK). This indicates that CHK binds to the same site on c-KIT to which FYN binds, possibly bringing the two into proximity on associated c-KIT subunits and leading to the down-regulation of FYN by CHK.

Animals↗

CDC2-related kinase PITALRE phosphorylates pRb exclusively on serine and is widely expressed in human tissues.

Mammalian cell cycle progression is regulated by sequential activation and inactivation of cyclin-dependent kinases (cdks). Recently, several new members of the cdk family were cloned, and some of these were shown to complex with different cyclins and to be active at discrete stages of the cell cycle. PITALRE, a new member of this family, was cloned by our laboratory and was shown to be able to phosphorylate pRb protein in vitro. In the current work, we found that PITALRE kinase activity phosphorylated pRb at sites similar to those phosphorylated by the CDC2 kinase, which itself is known to mimic, in vitro, the in vivo phosphorylation of pRb. Phosphorylation of pRb by the PITALRE-associated kinase activity was on Ser residues exclusively. Moreover, we investigated the expression pattern of PITALRE in normal human tissues, using immunohistochemical techniques so as to gain additional data on the characteristics of this new cdk family member. The protein was widely expressed, although a different tissue distribution and/or level of expression was found in various organs. Some specialized tissues such as blood, lymphoid tissue, ovarian cells, and the endocrine portion of the pancreas showed a high expression level of PITALRE. The specific expression pattern found suggests that PITALRE may be involved in specialized functions in certain cell types.

Antibodies↗

Angiotensinogen gene variation in a population case-control study of preeclampsia/eclampsia in Australians and Chinese.

Preeclampsia/eclampsia (PE/E) is a common disease of human pregnancy with a strong genetic component. The etiology of PE/E is unknown. Two recent reports indicated that the angiotensinogen gene (AGT) could be involved in susceptibility to PE/E. We performed a population-based case-control study in Australian and Chinese populations to investigate whether AGT is a good candidate gene for PE/E. A microsatellite polymorphism within AGT was typed as well as a molecular variant T235 (Met-->Thr) of AGT using allele-specific PCR and allele-induced restriction site PCR. The allele distributions of the microsatellite and the variant T235 of AGT were significantly different between the two ethnic groups. However, no significant allele associations were found with disease when comparing PE/E patients and controls in Australian or Chinese populations, which is in contrast to the two earlier reports. The results suggest that the contribution of AGT to the occurrence of PE/E is small, if anything, and is not constant across populations.

Alleles↗