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Biomedical subjects

L Sun

Publications and source records attributed to L Sun.

At least 343 records · Page 19Linked to original sources

A Myb-related transcription factor is involved in the phytochrome regulation of an Arabidopsis Lhcb gene.

We have isolated the gene for a protein designated CCA1. This protein can bind to a region of the promoter of an Arabidopsis light-harvesting chlorophyll a/b protein gene, Lhcb1*3, which is necessary for its regulation by phytochrome. The CCA1 protein interacted with two imperfect repeats in the Lhcb1*3 promoter, AAA/cAATCT, a sequence that is conserved in Lhcb genes. A region near the N terminus of CCA1, which has some homology to the repeated sequence found in the DNA binding domain of Myb proteins, is required for binding to the Lhcb1*3 promoter. Lines of transgenic Arabidopsis plants expressing antisense RNA for CCA1 showed reduced phytochrome induction of the endogenous Lhcb1*3 gene, whereas expression of another phytochrome-regulated gene, rbcS-1A, which encodes the small subunit of ribulose-1,5-bisphosphate carboxylase/oxygenase, was not affected. Thus, the CCA1 protein acts as a specific activator of Lhcb1*3 transcription in response to brief red illumination. The expression of CCA1 RNA was itself transiently increased when etiolated seedlings were transferred to light. We conclude that the CCA1 protein is a key element in the functioning of the phytochrome signal transduction pathway leading to increased transcription of this Lhcb gene in Arabidopsis.

Amino Acid Sequence↗

Transfection with aFGF cDNA improves wound healing.

Somatic gene therapy is a potentially useful strategy for the delivery of growth factors or cytokines to enhance wound healing. Experimental excisional and incisional wounds in impaired-healing diabetic mice (db/db) were treated with aFGF and with a plasmid coding for aFGF. A eukaryotic expression plasmid composed of the Hst signal peptide sequence in-frame with the human aFGF sequence was used. Transfection of tissues was accomplished either by direct plasmid uptake or by uptake facilitated with cationic liposomes. The results show that the closure of excisional wounds was significantly accelerated (p < 0.05) by topical application of human recombinant aFGF or by transfection with the aFGF plasmid but not by vehicle or control plasmid not containing the aFGF sequence. In incisional wounds, aFGF or transfection with the plasmid significantly increased the wound-breaking strength compared to their corresponding controls (p < 0.05). Quantitative histology of the plasmid-treated incisional wound sections revealed improved wound quality. The transcription of mRNA from human aFGF cDNA in the incisional wound tissue extracts was confirmed by RT-PCR, and the expressed aFGF was detected by immune dot blot and immunohistochemistry assays. The transfection was a transient process with a peak at 9 d in db/+ (littermates of the diabetic mice) incisional wounds, at 36 d in db/db incisional wounds, and at 27 d in db/db excisional wounds. Cells transfected with human aFGF occupied up to 6.4% of the transectional area in the wound sites. Thus, aFGF gene delivery resulted in both gene expression and a functional improvement in healing.

Animals↗

Increased DNA methyltransferase expression is associated with an early stage of human hepatocarcinogenesis.

The present study was designed to determine whether changes in DNA methyltransferase (DNA MTase) expression are involved in hepatocarcinogenesis. We examined DNA MTase expression in normal liver tissue (with no remarkable histological findings), liver tissue showing chronic hepatitis or cirrhosis, which are generally thought to be precancerous conditions, and hepatocellular carcinomas (HCCs) using the reverse-transcriptase polymerase chain reaction assay. DNA MTase mRNA levels were significantly higher in liver tissue showing chronic hepatitis and cirrhosis (DNA MTase mRNA/beta-actin mRNA ratio = 0.30 +/- 0.22, n = 24, P < 0.01) than in normal liver tissue either from patients with liver metastatic lesions of colonic cancer (0.14 +/- 0.05, n = 6) or from patients with HCCs (0.16 +/- 0.07, n = 3). DNA MTase mRNA levels were even higher in HCC tissue (0.34 +/- 0.18, n = 29). These results suggest that increased DNA MTase expression may be an early event during hepatocarcinogenesis. DNA MTase is a potential target for HCC preventive therapy.

Carcinoma, Hepatocellular↗

Human cytomegalovirus US2 destabilizes major histocompatibility complex class I heavy chains.

Human cytomegalovirus (HCMV) infection causes down-regulation of major histocompatibility complex class I heavy chains. We determined previously that there are two HCMV loci which encode functions responsible for that phenotype and that US11 is one of these loci (T. R. Jones, L. A. Hanson, L. Sun, J. S. Slater, R. M. Stenberg, and A. E. Campbell, J. Virol. 69:4830-4841, 1995). Through the construction and analysis of defined viral mutants and stably transfected cell lines, we identify US2 as the other locus. US2 is expressed from very early through late times postinfection, with its predominant product being a relatively unstable 24-kDa endoglycosidase H-resistant glycoprotein. In cell lines constitutively expressing US2, free class I heavy chains are proximal targets for US2-induced degradation, shortly after their synthesis. Both US2 and US11 can function in concert with US3 to down-regulate class I. Beta-2-microglobulin-associated heavy chains which are retained in the endoplasmic reticulum as a result of binding to the US3 glycoprotein are susceptible to destabilization caused by both US2 and US11 gene products. Thus, three HCMV genes which affect either the stability or the transport of class I heavy chains have been identified. The observation that each of these proteins is most abundant early in the replicative cycle suggests that they may play an important immunomodulatory role in vivo prior to productive infection, either during the latent or persistent phase or during reactivation.

Antigens, Viral↗

Effects of praeruptorine C on the intracellular free calcium in normal and hypertrophied rat ventricular myocytes.

AIM: To study the intracellular free calcium ([Ca2+]i) in normal and hypertrophic left ventricular myocytes isolated from adult rat hearts and the effects of praeruptorine C (Pra-C) on them. METHODS: [Ca2+]i of single myocyte was measured with Fura 2-AM. RESULTS: The resting [Ca2+]i was 87 +/- 4 nmol.L-1 in normal left ventricular myocytes, 123 +/- 7 nmol.L-1 in hypertrophied myocytes. After exposure to KCl (20, 40, and 60 mmol.L-1), the [Ca2+]i were increased by 66%, 141%, and 268% in normal myocytes, and 77%, 185%, and 243% in hypertrophic myocytes, respectively. Pra-C (1, 10, and 100 mumol.L-1) concentration-dependently inhibited the [Ca2+]i elevation caused by KCl (35 mmol.L-1) or norepinephrine (20 mumol.L-1) in both normal and hypertrophied myocytes. All of the effects of Pra-C were similar to that of nifedipine. CONCLUSION: [Ca2+]i of hypertrophied myocytes was higher than that of normal ones and Pra-C decrease the [Ca2+]i elevation in left ventricular myocytes resulted from its calcium channel blockade.

Animals↗

[Protection of hepatocyte growth factor against carbon tetrachloride injury in primary rat hepatocyte culture].

The protective effect of recombinant human hepatocyte growth factor (r-hHGF) against carbon tetrachloride (CCl4) injury in cultured rat hepatocytes was investigated. It was found that r-hHGF had cytoprotective effect on hepatocytes anainst CCl4 poisoning. Pretreatment with r-hHGF significantly increased the viability of cultured hepatocytes, reduced the leakage of intracellular alanine transaminase and potassium into the culture medium, as compared with the CCl4 group. The results suggest that r-hHGF may protect hepatocytes against CCl4 injury through lessening the severity of membrane lesion, preventing intracellular alanine transaminase and potassium leakage and sustaining the integrity of hepatocyte membrane.

Animals↗

[Effects of both puerarin and gypsum on the firing of pyrogen-treated thermosensitive neurons in the region POAH of anesthetized cats].

To investigate the possible central mechanism of antipyretic effects of Chinese medicines, puerarin and gypsum, the firing rate of thermosensitive neurons in preoptic-anterior hypothalamus (POAH) region of 34 cats was recorded by using extracellular micro-electrode technigue. Injection of pyrogen caused decrease in firing rate of 14 warm-sensitive neurons and increase in firing rate of 11 cold-sensitive neurons in the region of POAH. The effects could be reversed by the injection of puerarin and gypsum. The results showed the antipyretic action of puerarin and gypsum might be mediated by influences on the electroactivity of pyrogen-treated thermosensitive neurons in the region of POAH. Effect of puerarin and gypsum on cold-sensitive neurons was stronger than that of single one, suggesting that both are synergic in central level.

Action Potentials↗

[Chemical constituents of fructus Amomi].

Seven compounds have been isolated from Fructus Amomi. They were identified as bornyl acetate, camphor, borneol, beta-sitosterol, vanillic acid, stearic acid and palmitic acid. The vanillic acid was found in this medicine for the first time. Fifty-seven components were identified from the volatile oil of Fructus Amomi on the basis of GC-MS analysis. Eight components in the volatile oil over 1% in content are bornyl acetate, camphor, borneol, limonene, camphene, myrecene, carene-3 and alpha-terpeneol.

Drugs, Chinese Herbal↗

[Effects of praeruptorin C on cardiac dysfunction, myocardial compliance and collagen content in renovascular hypertensive rats].

The effects of praeruptorin C (Pra-C) on cardiac dysfunction, myocardial compliance, and collagen content in renovascular hypertensive rats have been studied. In rats with left ventricular hypertrophy (LVH), the ratio of cardiac flow (CF)/heart wet weight (HWW) and cardiac output (CO)/HWW decreased by 20.1% and 35.5%; LVSP and -dp/dtmax decreased by 19.4% and 42.1%, while LVEDP and T value were increased by 186.4% and 226%. The left ventricular diastolic pressure-volume (P-V) curve was shifted to the left and down ward compared with that of sham-operated rats. The hydroxyproline content was 5.93 +/- 0.82 mg.g-1 dry weight in the left ventricular myocardial tissue of rats with LVH which was higher than that of sham-operated rats(e.g. 4.01 +/- 0.47 mg.g-1). The content did not change in the right ventricular myocardium. After treated with Pra-C for nine weeks, the CF/HWW and CO/HWW increased by 31.3% and 25.1%, the LVSP and -dp/dtmax increased by 16.7% and 27.8%. The LVEDP and T value was reduced by 34.9% and 36.5%, respectively, compared with those of LVH. The P-V curves of these rats were shifted to the right and in the up-ward direction. The hydroxyproline content was 4.52 +/- 0.3 mg.g-1 dry weight in the left ventricular tissue. These results indicate that Pra-C improved the cardiac contractility and relaxation by increasing the coronary flow and cardiac output, and decreasing the collagen content in the ventricular myocardium.

Animals↗

[Studies on the chemical constituents of Annona muricata].

Annonaceous acetogenin (or polyketide) is a kind of potential antineoplastic agents from Annonaceae plants. Two new acetogenins, Muricatalicin (I) and muricatalin (VI), a mesitoate of a new acetogenin, annonacin-B mesitoate (Vb), and three known acetogenins, annonacin (II), annonacin-A (III) and annonacin-10-one (IV) have been isolated from Annona muricata L. The structures and relative stereochemistry of I, VI and Vb were elucidated on the basis of spectral analysis and examination of their acetates and/or mesitoate.

Annonaceae↗

[Cloning and permanent expression in eukaryotic cells of rhTpo].

OBJECTIVE: To obtain recombinant human thrombopoietin (rhTpo). METHODS: rhTpo gene was cloned from fetal liver mRNA by means of RT-PCR and subcloned into permanent expressing plasmid RC/CMV in order to form a recombinant permanent expressing plasmid RC/CMV/Tpo. RESULTS AND CONCLUSION: After being transferred into CHO cells, the eukaryotic permanent expression of rhTpo, which can enhance murine megakaryocyte colony growth in vitro, was obtained.

Animals↗

Role of alpha 5 beta 1 integrin in determining malignant properties of colon carcinoma cells.

We characterized the expression of alpha 5 beta 1 integrin in two distinct phenotypes of colon carcinoma cell lines. Highly invasive colon cell lines (designated Group I cell lines) expressed higher levels of integrin alpha 5 beta 1 mRNA and protein than did poorly invasive colon cell lines (designated Group III cell lines). The relatively high expression of integrin alpha 5 beta 1 in Group I cell lines resulted in strong enhancement of cell adhesion to fibronectin (FN) tissue culture plates, whereas Group III cell lines showed little or no enhancement of cell adhesion by coating. There was no significant difference between Group I and Group III cell lines with respect to cell adhesion to laminin and collagen IV. Cell adhesion to FN in Group I cells was mainly mediated by integrin alpha 5 beta 1 because a monoclonal anti-alpha 5 subunit antibody could block cell adhesion to FN, whereas anti-alpha 2 and anti-alpha 3 antibodies had no effect on cell adhesion to FN. The divergence of alpha 5 beta 1 expression in these two distinct colon carcinoma phenotypes suggested that high expression of alpha 5 beta 1 might contribute to malignant progression in this model system. To test this hypothesis, GEO cells, a Group III cell line that did not express alpha 5 integrin, were transfected with the alpha 5 subunit. Stable transfection of alpha 5 sense cDNA into a typical GEO-limiting dilution clone led to the expression of alpha 5 subunit mRNA and cell surface alpha 5 beta 1 protein. The alpha 5 sense transfectants showed enhanced attachment to FN-coated plates and were more tumorigenic when the cells were injected into athymic nude mice. These results indicate that inappropriately high alpha 5 beta 1 integrin expression contributes to malignant progression in colon carcinoma.

Animals↗

[Repeated low-flow sevoflurane anesthesia: effects on hepatic and renal function in beagles].

We studied the effects of repeated low-flow sevoflurane anesthesia for 6 hours. Five beagle dogs received 1.3 MAC (3%) sevoflurane anesthesia. Anesthesia of 6 hours was repeated on at the 7th day after the first anesthesia. Compound A gas samples were collected from the inspiratory limb during anesthesia. Concentrations of serum and renal fluoride, hepatic and renal function parameters were measured during and up to 7 days after the first and second anesthesia. The peak concentration of compound A was 23.7 +/- 3.6 ppm at 2 hours and the same level remained during the anesthesia. Plasma fluoride level exceeded 50 mmol.l-1 during anesthesia and rapidly decreased to the preanesthesia level thereafter. Serum GOT increased slightly only on the first postanesthesia day. No significant changes in other blood chemistry studies were observed. The excretion of renal tubular enzymes did not increase during and after anesthesia. Repeated low flow sevoflurane anesthesia in beagles did not affect hepatic and renal function significantly.

Acetylglucosaminidase↗

An evaluation of minimal residual disease in childhood acute lymphoblastic leukaemia.

Evaluation of minimal residual disease (MRD) in acute lymphoblastic leukaemia (ALL) is important for disease prognostication and early relapse detection. In this study, the rearranged third complementarity-determining-region (CDR-III) of immunoglobulin heavy chain (IgH) was used as a surrogate tumour marker for MRD evaluation. DNA obtained from marrows at diagnosis was amplified by the polymerase chain reaction (PCR) using a pair of consensus primers. After 2 rounds of DNA amplification and polyacrylamide gel separation, the nucleotide sequences of 87.5% (21/24) consecutive children with B-lineage ALL were obtained by automated sequencing. There were between 1-4 rearrangements per patient. Although the J5 and J6 joining regions were preferentially used, the rearranged sequences were unique for all 25 sequences obtained. Oligoprobes to the DNJ region were constructed and quantitation in 7 patients showed a detection sensitivity of 1 leukaemic cell in 10(4) to 10(5) normal cells compared to 3 in 100 using conventional morphological criteria. Serial bone marrow showed progressive decrease in the quantity of leukaemic cells, and no leukaemic sequences were detected during cessation of therapy in 4/7 patients. One patient with detectable MRD, absconded treatment and eventually relapsed. These results are consistent with the need to eliminate the leukaemic clones below MRD detection levels before the end of therapy at 2 years. In conclusion, this study describes a novel, simplified and sensitive method of MRD detection in childhood leukaemia.

Adolescent↗

Autologous bone marrow transplantation in a child with acute promyelocytic leukemia in second remission.

Acute myeloid leukemia (AML) comprises 15%-20% of childhood acute leukemia cases. The long-term disease free survival (DFS) in childhood AML is poor with standard chemotherapy alone. Early intensive chemotherapy is generally regarded to be necessary for achieving high complete remission (CR) rates. Recent experience has shown that incorporation of early intensification with high-dose melphalan conditioning and autologous bone marrow transplantation (BMT) during the first remission significantly improves long-term DFS in children with AML. In this article, we report the use of autologous BMT for treatment of a three-and-half year old child with acute promyelocytic leukemia (APL or M3) in second remission. The patient was conditioned with high-dose melphalan of 180 mg/kg prior to bone marrow reinfusion. A total of 4.0 x 10(7)/kg mononuclear cells and 1.07 x 10(5)/kg granulomonocytic colony forming units (CFU-GM) were infused. Haematopoietic stem cells were enriched by almost 20-fold after the separation and cryopreservation procedures. Haematological recovery was achieved four-and-a-half weeks post-BMT. She has remained in complete remission 18 months after transplantation. Our experience in this patient indicates that this procedure can be used in second remission and it may provide a better alternative for the management of childhood AML in Singapore.

Antineoplastic Agents, Alkylating↗

Down-regulation of cytokine expression in murine lymphocytes by PACAP and VIP.

Neuropeptides, such as VIP and PACAP, released or produced in the microenvironment of the primary and secondary lymphoid organs, could affect a variety of immune responses through the regulation of cytokine expression. VIP has been previously shown to inhibit IL-2, IL-4, and IL-10 production in murine lymphocytes stimulated through the TCR-associated CD3 complex. This study shows that, similar to VIP, PACAP-38 inhibits IL-2 production in T lymphocytes. Comparisons with forskolin, a known cAMP inducer, suggest that the increase in intracellular cAMP represents at least one of the transduction pathways involved in IL-2 inhibition, especially in the higher range of neuropeptide concentration. Studies of the detailed molecular mechanisms involved in the regulation of IL-2 expression indicate that reduction of de novo transcription and destabilization of the message contribute to the reduction of steady-state IL-2 mRNA levels following VIP treatment. Examination of several IL-2 transcriptional factors indicates that only NFAT is down-regulated by VIP. Neuropeptides, such as VIP and PACAP, which specifically modulate the expression of various cytokines, could play an important role in the intricate cytokine network controlling local immune responses.

Animals↗