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Biomedical subjects

J Ye

Publications and source records attributed to J Ye.

At least 199 records · Page 11Linked to original sources

Negative regulation of cytokine gene transcription.

Cytokines are a class of soluble proteins that mediate signals throughout the immune system as well as between immune effector cells and other cell populations. This class of proteins includes interleukins, interferons, and chemokines. The expression of cytokines is tightly controlled in the producing cells, and one of the most important regulatory steps in this control is gene transcription. The transcription of most cytokine genes is silent until a producing cell is activated by extracellular stimuli. It is very common that transcription of these genes initiates immediately upon activation and shuts down quickly even in the continuous presence of the stimulating agent. Work performed over the past decade has revealed much about the molecular mechanism responsible for transcriptional regulation of these genes. Positive and negative transcription factors function in a concerted manner to regulate transcription of a specific cytokine at the promoter or intron region. We focus on recent progress in the field of transcriptional regulation of cytokine genes with an emphasis on the negative regulation of cytokine gene transcription.

Animals↗

Myocardial ischemia decreases oxidative phosphorylation through cytochrome oxidase in subsarcolemmal mitochondria.

The effect of myocardial ischemia on mitochondrial oxidative phosphorylation was investigated using isolated, buffer-perfused rabbit hearts. After 45 min of global ischemia, oxidative phosphorylation was decreased only in the subsarcolemmal population of mitochondria with all substrates tested. The oxidation of N,N,N',N' tetramethyl p-phenylenediamine-ascorbate, an electron donor to cytochrome oxidase via cytochrome c, was decreased in subsarcolemmal mitochondria [ischemia (n = 6): 76 +/- 3 vs. control (n = 5): 105 +/- 6 nanoatoms O.min-1.mg-1, P < 0.01] but not in interfibrillar mitochondria. Only minor morphological changes were observed by electron microscopy in the isolated mitochondria after ischemia. Neither cytochrome oxidase activity measured under conditions for maximal activity nor the apparent Michaelis constant and maximum velocity values of the two cytochrome c binding sites were different in subsarcolemmal mitochondria isolated from ischemic and control hearts. The cytochrome c content was decreased in subsarcolemmal mitochondria after ischemia (ischemia: 0.111 +/- 0.013 vs. control: 0.156 +/- 0.007 nmol/mg protein, P < 0.05). Thus ischemia decreased the rate of oxidative phosphorylation through cytochrome oxidase selectively in intact subsarcolemmal mitochondria. Ischemic damage to the terminal segment of the electron transport chain involves a decrease in the content of cytochrome c, whereas the expressible catalytic activity of cytochrome oxidase remains unchanged.

Adenosine Diphosphate↗

[Experimental study of anti-tumour effect of rhTNF alpha on the laryngeal squamous cell carcinoma].

The anti-tumor effect of rhTNF alpha (recombination human tumor necrosis factor) with three different doses on laryngeal squamous cancer was investigated in nude mice. The results indicated that the higher and the middle doses of rhTNF alpha had significant antitumour effect against laryngeal cancer in BALB/C nude mice. The inhibitory rate of tumor growth was 54.4%(P < 0.05). The lower dose of rhTNF alpha also had antitumour effect but the cancer grew again when rhTNF alpha was stopped. Ultrastructural changes showed that the plasma membrane and the nucleus envelope of the tumor cell were destroyed and the number of lysosomes increased, fibroid material could be seen in the cancer capillary.

Animals↗

[The development of an computerized analysis system for the women pelvic cavity impedance rheogram].

This article introduces the components and functions of an computered analysis system for the women pelvic cavity impedance rheogram. It can sample the pelvic cavity impedance rheogram signal of the body double-side and the reference ECG signal. With help of manual intervence, it also can automatically recognize characteristic points of the pelvic cavity impedance rheogram and measure the parameters. It can correct the recognized characteristic points simultaneously and print the pelvic cavity impedance rheogram of the body double-side and 15 characteristic data parameters. After clinical applying, the analysis results of mors 100 patients make clear that: this system has remarkably clinical significance for diagnosis of the women pelvic cavity extravasated blood and evaluation of treating effect, so it has much better clinical applied expectation.

Equipment Design↗

[In vitro infection of the human T-cell line with the hepatitis C virus].

A human T-cell line (Molt-4) was infected with an hepatitis C virus RNA (HCV- RNA) positive serum for 12 hours and incubated continously for 1-28 days. A productive infection was demonstrated by polymerase chain reaction amplification. Viral plus-stranded RNA was detectable in Molt-4 on the 1st, 2nd, 3rd day and the 8th day post infection. Viral minus-stranded RNA was detectable on the 2nd day and 8th day post infection.

Cell Line↗

A new technique of coronary artery ligation: experimental myocardial infarction in rats in vivo with reduced mortality.

In vivo models of myocardial infarction following coronary artery ligation in the rat still suffer from high early mortality and a low rate of success of myocardial infarction. This study investigated the possibility of reducing early mortality and increasing the rate of myocardial infarction by modifications of surgical techniques. Eighteen rats were divided into two groups: normal control (3 rats) and ligation (15 rats). The major modifications of surgical techniques used in this study include: (1) no exteriorization of the heart, (2) ligation of the origins of the branches rather than the main trunk of the left coronary artery, (3) removal of air from the chest after closure, (4) supplying oxygen immediately after extubation. Following surgery, the rats recovered uneventfully and 11 rats were alive after 16 weeks. One rat, with a large myocardial infarction, died 2 h after surgery. Early mortality (during surgery and 1 week after surgery) was 6.7% with a success rate of myocardial infarction of 85%. The left ventricle in the ligation group showed significant dilation relative to normal and sham-operated control hearts (317% of control hearts, p < 0.001). However, myocardial mass did not increase. The average infarct size was 33%. These results demonstrate that a reduction in early mortality and an increased success rate of myocardial infarction can be achieved by modifications of surgical techniques.

Analysis of Variance↗

Biological and clinical significance of cytogenetic study on 100 acute lymphoblastic leukemia and 219 acute non-lymphoblastic leukemia.

OBJECTIVE: To further investigate the role that cytogenetic may play in the diagnosis and prognosis of leukemia, a study was conducted in 319 acute leukemias. METHODS: 100 patients with acute lymphoblastic leukemia (ALL) and 219 patients with acute non-lymphoblastic leukemia (ANLL) were from Rui Jin Hospital, Xin Hua Hospital, Ren Ji Hospital and Shanghai Children's Hospital. Their cytogenetic data were analyzed together with those of morphology, immunology and clinical prognosis. RESULTS: In ALL group, 48 cases were karyotypically normal whereas 52 cases revealed chromosomal changes, among which 32 had quantitative abnormalities and 20 had qualitative abnormalities. The translocation t(9; 22) was identified in 11 out of 20 cases of structural aberrations (55%). Specific structural aberrations t(9; 22) and t(8; 14) were detected to be related to B-lineage associated differentiation antigens and t(8; 14) also with ALL-L3 according to FAB classification. With regard to clinical prognosis, the survival rate of structural aberration subset decreased significantly compared with the normal karyotype subset (P < 0.05). However, no statistically significant difference was found between hyperdiploidy subset (not including near-triploidy) and normal karyotype subset (P > 0.75). In ANLL group, 80% of de novo patients and relapsed patients had chromosomal abnormalities. Importantly, structural aberrations accounted for 73% of these abnormalities and frequently corresponded to specific types of FAB classification. Relevant prognostic studies demonstrated that t(15; 17) subset had the best overall survival probability, followed by t(8; 21) and normal karyotype subset, while the numerical aberration subset showed a relatively poor prognosis. CONCLUSION: Our data confirmed that cytogenetic study is important for the molecular study of the leukemogenesis. On the other hand, it also provides an independent parameter for prognosis in acute leukemia.

Adolescent↗

Serological investigation on human trichinellosis spiralis in Hubei Province of PR China.

PVC-ELISA for detection of human trichinellosis and its value of field application was explored during 1988-1994. Sera of 56 cases of clinically confirmed trichinellosis were tested for serum Trichinella antibodies by way of PVC-ELISA, and the positive rate was 100%. When the sera of 80 cases of healthy persons from non-endemic areas were tested for trichinellosis, they were all negative. For the purpose of exploring the cross-reaction of PVC-ELISA used for detection of trichinellosis, sera from 144 cases of other parasitoses were tested. Among them, the sera of 80 cases of schistosomiasis japonica for trichinellosis were negative; 30 cases of paragonimiasis and 30 cases of clonorchiasis were also negative. Among sera of 4 cases of cysticercosis, there was only one positive. The results showed that PVC-ELISA for trichinellosis had a high sensitivity and specificity. For field application, a population of 800 from endemic areas were examined by PVC-ELISA, 85 (10.63%) were positive. The infected persons were mainly in Xiangfan region of Hubei Province. The percentage of the positive of trichinellosis in Xiangfan region was 64.71% (55/85). The positive rate of males and females was 13.41% (57/425) and 7.47% (28/375), respectively. There was a difference among the age groups, with the highest in those > or = 20 years. The findings of this field investigation suggested that the infection rate of trichinellosis spiralis was high in this province, especially in the Xiangfan region; and that PVC-ELISA was suitable for field application. The emphasis of trichinellosis control and research of this province should be in the Xiangfan region.

China↗

Retinoic acid-induced transcriptional modulation of the human interferon-gamma promoter.

Disregulation of vitamin A metabolism is able to generate different immunological effects, including altered response to infection, reduced IgG production, and differential regulation of cytokine gene expression (including interleukin-2 and -4 and interferon-gamma (IFN-gamma)). In particular, IFN-gamma gene expression is significantly affected by vitamin A and/or its derivatives (e.g. retinoic acid (RA)). Here, we analyze the effect of retinoic acid on IFN-gamma transcription. Transient transfection assays in the human T lymphoblastoid cell line Jurkat demonstrated that the activation of the IFN-gamma promoter was significantly down-regulated in the presence of RA. Surprisingly, two different AP-1/CREB-ATF-binding elements situated in the initial 108 base pairs of the IFN-gamma promoter and previously shown to be critical for transcriptional activity were unaffected by RA. Utilizing promoter deletions and electrophoretic mobility shift analysis, we identified a USF/EGR-1-binding element cooperating in the modulation of IFN-gamma promoter activity by RA. This element was found to be situated in a position of the IFN-gamma promoter close to a silencer element previously identified in our laboratory. These results suggest that direct modulation of IFN-gamma promoter activity is one of the possible mechanisms involved in the inhibitory effect of retinoids on IFN-gamma gene expression.

Humans↗

Acylation of glucosaminyl phosphatidylinositol revisited. Palmitoyl-CoA dependent palmitoylation of the inositol residue of a synthetic dioctanoyl glucosaminyl phosphatidylinositol by hamster membranes permits efficient mannosylation of the glucosamine residue.

Two critical steps in the assembly of yeast and mammalian glycosylphosphatidylinositol (GPI) anchor precursors are palmitoylation of the inositol residue and mannosylation of the glucosamine residue of the glucosaminyl phosphatidylinositol (GlcNalpha-PI) intermediate. Palmitoylation has been reported to be acyl-CoA dependent in yeast membranes (Costello, L. C., and Orlean, P. (1992) J. Biol. Chem. 267, 8599-8603) but strictly acyl-CoA independent in rodent membranes (Stevens, V. L., and Zhang, H. (1994) J. Biol. Chem. 269, 31397-31403), and thus poorly conserved. In addition, it was suggested that acylation must precede mannosylation in both yeast (Costello, L. C., and Orlean, P. (1992) J. Biol. Chem. 276, 8599-8603) and rodent (Urakaze, M., Kamitani, T., DeGasperi, R., Sugiyama, E., Chang, H.-M., Warren, C. D., and Yeh, E. T. H. (1992) J. Biol. Chem. 267, 6459-6462) cells because GlcNalpha-acyl-PI accumulates in vivo when mannosylation is blocked. However, GlcNalpha-acyl-PI accumulation would also be expected if mannosylation and acylation were independent of each other. These issues were addressed by the use of a synthetic dioctanoyl GlcNalpha-PI analogue (GlcNalpha-PI(C8)) as an in vitro substrate for GPI-synthesizing enzymes in Chinese hamster ovary cell membranes. GlcNalpha-PI(C8) was acylated in an manner requiring acyl-CoA. Thus, the process involving acyl-CoA reported for yeast has been conserved in mammals. Furthermore, both GlcNalpha-PI(C8) and GlcNalpha-acyl-PI(C8) could be mannosylated in vitro, but mannosylation of the latter was significantly more efficient. This provides direct support for the earlier suggestion that acylation precedes mannosylation in rodents cells. A similar result was also observed with the Saccharomyces cerevisiae mannosyltransferase. In contrast, it has been reported that mannosylation of endogenous GlcNalpha-PI by Trypansoma brucei membranes occurs without prior acylation. The same result was obtained with GlcNalpha-PI(C8), confirming that the mannosyltransferase of trypanosomes is divergent from those in yeasts and rodents.

Acyl Coenzyme A↗

RIG-E, a human homolog of the murine Ly-6 family, is induced by retinoic acid during the differentiation of acute promyelocytic leukemia cell.

In vivo all-trans-retinoic acid (ATRA), a differentiation inducer, is capable of causing clinical remission in about 90% of patients with acute promyelocytic leukemia (APL). The molecular basis for the differentiation of APL cells after treatment with ATRA remains obscure and may involve genes other than the known retinoid nuclear transcription factors. We report here the ATRA-induced gene expression in a cell line (NB4) derived from a patient with APL. By differential display-PCR, we isolated and characterized a novel gene (RIG-E) whose expression is up-regulated by ATRA. The gene is 4.0 kb long, consisting of four exons and three introns, and is localized on human chromosome region 8q24. The deduced amino acid sequence predicts a cell surface protein containing 20 amino acids at the N-terminal end corresponding to a signal peptide and an extracellular sequence containing 111 amino acids. The RIG-E coded protein shares some homology with CD59 and with a number of growth factor receptors. It shares high sequence homology with the murine LY-6 multigene family, whose members are small cysteine-rich proteins differentially expressed in several hematopoietic cell lines and appear to function in signal transduction. It seems that so far RIG-E is the closest human homolog of the LY-6 family. Expression of RIG-E is not restricted to myeloid differentiation, because it is also present in thymocytes and in a number of other tissues at different levels.

Amino Acid Sequence↗

The transition of pre-BI to pre-BII cells is dependent on the VH structure of the mu/surrogate L chain receptor.

We have demonstrated previously that the majority ( > 90%) of VH12 B cells are absent from the adult peripheral repertoire, and that most that remain have the fourth position at the D-J function (designated 10/G4). We report here that most VH 12-expressing pre-B cells are lost during the transition from the pre-BI to the pre-BII cell stage in normal mice, and that pre-BII cell productive (P) rearrangements ar enriched in 10/G4 CDR3. This coincides with the initial expression of H chain and the generation of the mu/surrogate L chain (SL) receptor. In contrast, there is not enrichment for 10/G4 CDR3 in mu MT mice, and the frequency of P rearrangements is as expected from a random rearrangement mechanism, ruling out a biased rearrangement mechanism unique to VH12. We have also demonstrated that non-10/G4 mu chains can associate with SL and be expressed on the cell surface, suggesting that they are available on the cell surface for selection. Thus, transition of pre-BI to pre-BII cells is dependent on the structure of the VH domain.

Animals↗

Creatine phosphate as the preferred early indicator of ischemia in muscular tissues.

Changes in creatine compounds, especially the creatine phosphate to creatine ratio (CrP/Cr), are more sensitive indicators than changes in other metabolites for early ischemia in the different muscular tissues of heart, small intestine, skeletal muscle, and aorta. Changes in adenine nucleotide ratios are buffered by CrP reserves and the absolute concentration of adenine nucleotides can vary greatly between different muscular tissues. Accumulation of lactate is indicative of ischemia, but is not as sensitive as the ratio of CrP/Cr, but may better indicate the duration of ischemia. Glycerol also accumulates in muscular tissues during prolonged ischemia, so that consideration of both lactate and glycerol levels together, might confer a better estimate of the duration of ischemia of different muscular tissues.

Adenine Nucleotides↗

Vanadium(IV)-mediated free radical generation and related 2'-deoxyguanosine hydroxylation and DNA damage.

Free radical generation, 2'-deoxyguanosine (dG) hydroxylation and DNA damage by vanadium(IV) reactions were investigated. Vanadium(IV) caused molecular oxygen dependent dG hydroxylation to form 8-hydroxyl-2'-deoxyguanosine (8-OHdG). During a 15 min incubation of 1.0 mM dG and 1.0 mM VOSO4 in phosphate buffer solution (pH 7.4) at room temperature under ambient air, dG was converted to 8-OHdG with a yield of about 0.31%. Catalase and formate inhibited the 8-OHdG formation while superoxide dismutase enhanced it. Metal ion chelators, DTPA and deferoxamine, blocked the 8-OHdG formation. Incubation of vanadium(IV) with dG in argon did not generate any significant amount of 8-OHdG, indicating the role of molecular oxygen in the mechanism of vanadium(IV)-induced dG hydroxylation. Vanadium(IV) also caused molecular oxygen-dependent DNA strand breaks in a pattern similar to that observed for dG hydroxylation. ESR spin trapping measurements demonstrated that the reaction of vanadium(IV) with H2O2 generated OH radicals, which were inhibited by DTPA and deferoxamine. Incubation of vanadium(IV) with dG or with DNA in the presence of H2O2 resulted in an enhanced 8-OHdG formation and substantial DNA double strand breaks. Sodium formate inhibited 8-OHdG formation while DTPA had no significant effect. Deferoxamine enhanced the 8-OHdG generation by 2.5-fold. ESR and UV measurements provided evidence for the complex formation between vanadium(IV) and deferoxamine. UV-visible measurements indicate that dG, vanadium(IV) and deferoxamine are able to form a complex, thereby, facilitating site-specific 8-OHdG formation. Reaction of vanadium(IV) with t-butyl hydroperoxide generated hydroperoxide-derived free radicals, which caused 8-OHdG formation from dG and DNA strand breaks. DTPA and deferoxamine attenuated vanadium(IV)/t-butyl-OOH-induced DNA strand breaks.

8-Hydroxy-2'-Deoxyguanosine↗

Preliminary report on treatment of bone tumors with microwave-induced hyperthermia.

Between July, 1992, and February, 1995, 62 patients with various bone tumors were treated with microwave-induced hyperthermia. The series had 47 cases of malignant tumors and 15 cases with benign tumors; most of the tumors occurred at or near knee joints (53/62 = 85.4%). The surgical procedure consisted of separating the tumorous segment from surrounding normal tissues with a safe margin, cooling the normal tissues (including the vital neurovascular bundle and the intrajoint structures) with a water circulation system while heating the tumor simultaneously with the microwave antenna array, and providing an adequate soft-tissue cover for the dead bone. The tumor core temperature and the surface temperature reached 108 and 65 degrees C, respectively. The duration of microwave irradiation was usually 40-50 minutes. Meanwhile, the temperature of the normal tissues was kept under 39 degrees C. The minimal and maximal periods of clinical observation were 3 months and 36 months, respectively, and the mean follow-up period was 17 months. The 62 cases were evaluated from both oncological and orthopedic points of view. Five cases had local recurrence and required amputation. The 57 other cases had excellent local control. Six malignancy cases died of lung metastasis during a period of 1-2 years. Pathological fracture occurred at devitalized bone in five cases. In most of the cases, the knee joints functioned well, were stable and painless, and had almost full range of motion. Single-photon emission-computed tomography study in 16 cases revealed that revascularization of the devitalized tumorous bone segment could be accomplished in 1 year or more. These results show that the use of microwave hyperthermia for the treatment of bone tumors can be considered to be a definitive operation procedure that is safe and is well tolerated by patients. The oncological and orthopedic results are very encouraging.

Adolescent↗

A recombinant peptide model of the first nucleotide-binding fold of the cystic fibrosis transmembrane conductance regulator: comparison of wild-type and delta F508 mutant forms.

A series of recombinant peptides, each including the sequence proposed to be the first nucleotide-binding fold of cystic fibrosis transmembrane conductance regulator (CFTR), has been produced in an attempt to find a model peptide that would autologously fold into a soluble structure with native-like properties. The peptide NBDIF, which contains the 267-amino acid sequence of CFTR from 384 to 650, meets these requirements. The peptide was produced with a high expression bacterial plasmid pRSET, purified from inclusion bodies following solubilization with 6 M guanidine-HCl and refolded from 8 M urea. Competitive displacement of trinitrophenol-ATP by nucleotides reveals binding of ATP and related nucleotides with KDs in the low micromolar range; the KD for ATP gamma S is 1.0 +/- 0.4 microM and for ADP 8.8 +/- 3.1 microM. The native-like character of the model peptide's structure is further supported by the findings that the KD for the ATP analog, 5'-adenylimidodiphosphate, is fourfold lower than the KD for the methylene analog, 5'-adenylmethylenediphosphonate, and that ATP binding slows the trypsin proteolysis of NBDIF. The CD spectra of NBDIF and the parallel peptide containing the most common cystic fibrosis mutation, deletion of Phe 508, are essentially indistinguishable, both spectra indicating 28% alpha-helix and 23% beta-sheet, with insignificant differences in the amounts of beta-turns and random structure. Extensive investigation using multiple conditions with highly purified preparations of the model peptides demonstrates that they do not support ATP hydrolysis. These large recombinant peptides offer practical models for the investigation of the first nucleotide-binding domain of CFTR.

Adenosine Diphosphate↗

Expression in Escherichia coli of cytoplasmic portions of the cystic fibrosis transmembrane conductance regulator: apparent bacterial toxicity of peptides containing R-domain sequences.

Large peptide segments (148-479 amino acids) of the cystic fibrosis transmembrane conductance regulator, which are projected to represent cytoplasmic portions of this large membrane protein, were expressed in Escherichia coli using two T7 RNA polymerase vectors, pET11a and pRSET. Five of the nine peptides were readily expressed at high levels (15-35 mg/liter) and one at an intermediate level (10 mg/liter), but three could not be expressed at >1.5 mg/liter regardless of efforts to further optimize the system. Preinduction testing of these latter plasmids failed to demonstrate any plasmid instability, while bacterial survival was drastically curtailed upon induction, beyond that observed with the other plasmids. Peptides containing the second half of exon 13 (residues 700-830; R domain) appear to be especially toxic to the expressing bacteria. Peptides including this hydrophilic segment may be inhibiting the bacterial permeases which are known to he homologous to other portions of CFTR.

Amino Acid Sequence↗