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Biomedical subjects

D Ren

Publications and source records attributed to D Ren.

At least 55 records · Page 3Linked to original sources

[Changes in chloroquine resistance of Plasmodium falciparum in Hainan province].

AIM: To observe the variation in resistance of Plasmodium falciparum after the cessation of chloroquine medication. METHODS: WHO standard in vitro microtest and in vivo test were used. RESULTS: In vitro test showed that the rate of chloroquine-resistant P. falciparum dropped from 97.9% in 1981 to 26.7% in 1997 (P < 0.001). The mean concentration of chloroquine for complete inhibition of schizont formation declined from 10.46 +/- 7.14 pmol/ul blood in 1981 to 1.63 +/- 1.47 pmol/microliter blood in 1997(P < 0.001). In vivo test showed that the rate of chloroquine-resistant P. falciparum decreased from 84.2% in 1981 to 18.4% in 1997(P < 0.001). The proportion of R III cases to the total resistant cases dropped from 53.1% in 1981 to 14.3% in 1997. CONCLUSION: A tendency of progressive decline of resistance of the parasite was revealed after the cessation of chloroquine medication.

Adolescent↗

Flower-associated brachycera flies as fossil evidence for jurassic angiosperm origins

Pollinating insects played a decisive role in the origin and early evolution of the angiosperms. Pollinating orthorrhaphous Brachycera fossils (short-horned flies) collected from Late Jurassic rocks in Liaoning Province of northeast China provide evidence for a pre-Cretaceous origin of angiosperms. Functional morphology and comparison with modern confamilial taxa show that the orthorrhaphous Brachycera were some of the most ancient pollinators. These data thus imply that angiosperms originated during the Late Jurassic and were represented by at least two floral types.

Journal Article↗

A mutation affecting dihydropyridine-sensitive current levels and activation kinetics in Drosophila muscle and mammalian heart calcium channels.

The Dmca1D gene encodes a Drosophila calcium channel alpha1 subunit. We describe the first functional characterization of a mutation in this gene. This alpha1 subunit mediates the dihydropyridine-sensitive calcium channel current in larval muscle but does not contribute to the amiloride-sensitive current in that tissue. A mutation, which changes a highly conserved Cys to Tyr in transmembrane domain IS1, identifies a residue important for channel function not only in Drosophila muscle but also in mammalian cardiac channels. In both cases, mutations in this Cys residue slow channel activation and reduce expressed currents. Amino acid substitutions at this Cys position in the cardiac alpha1 subunit show that the size of the side chain, rather than its ability to form disulfide bonds, affects channel activation.

Amiloride↗

Fibroblast growth factor-2-toxin induced cytotoxicity: differential sensitivity of co-cultured vascular smooth muscle cells and endothelial cells.

Recombinant FGF-2-SAP is a mitotoxin consisting of the plant-derived ribosome-inactivating toxin saporin (SAP) fused to basic fibroblast growth factor (FGF-2). FGF-2-SAP targets and kills cells bearing upregulated FGF receptors. In vivo, FGF-2-SAP inhibits smooth muscle cell hyperplasia in models of restenosis. The present study examined the potential for a differential effect of FGF-2-SAP on canine vascular endothelial cells (EC) and smooth muscle cells (SMC) separately as well as in a novel co-culture model. Canine vascular SMC and EC cultures were established separately and made quiescent once cells reached 80% confluence. Following the release from growth arrest, both cell types were treated with FGF-2-SAP, or FGF-2, or SAP alone for 48 h. [3H]TdR incorporation was used to determine the growth response of SMC and EC. The co-culture system was created by plating canine vascular SMC and EC on either side of a microporous 13 microm thick polyester membrane insert. Both cell types were grown to 80% confluence and independently made quiescent. Following the release from growth arrest, cells were treated with FGF-2-SAP, or FGF-2, or SAP alone. Negative and positive control groups were untreated wells containing phosphate buffered saline and complete growth media, respectively. After 48 h, both [3H]TdR incorporation and total DNA content, by fluorometric measurement, were quantitated in SMC and EC independently. FGF-2-SAP showed a concentration-dependent cytotoxicity in both canine SMC and EC but cytotoxicity for EC required substantially higher concentrations. In co-cultured SMC, FGF-2-SAP significantly decreased both [3H]TdR uptake and total DNA content at 0.5, 5, 50, and 500 ng/ml (0.01-10 nM) compared to positive controls. In co-cultured EC, FGF-2-SAP decreased [3H]TdR uptake at 50 and 500 ng/ml and total DNA content at 500 ng/ml compared to positive controls. Neither SAP alone nor FGF-2 alone showed a significant effect on [3H]TdR uptake or DNA content of either SMC or EC. In this unique co-culture model, which better replicates the relationship between SMC and EC in vivo, we demonstrated a dose-response range of FGF-2-SAP at which both the proliferation and total cell number of SMC, but not EC, is significantly reduced. These data suggest that FGF-2-SAP may have therapeutic utility in inhibiting myointimal hyperplasia in the absence of a deleterious effect on regenerating endothelium following vascular reconstructions.

Animals↗

Genetic and developmental characterization of Dmca1D, a calcium channel alpha1 subunit gene in Drosophila melanogaster.

To begin unraveling the functional significance of calcium channel diversity, we identified mutations in Dmca1D, a Drosophila calcium channel alpha1 subunit cDNA that we recently cloned. These mutations constitute the l(2)35Fa lethal locus, which we rename Dmca1D. A severe allele, Dmca1D(X10), truncates the channel after the IV-S4 transmembrane domain. These mutants die as late embryos because they lack vigorous hatching movements. In the weaker allele, Dmca1D(AR66), a cysteine in transmembrane domain I-S1 is changed to tyrosine. Dmca1D(AR66) embryos hatch but pharate adults have difficulty eclosing. Those that do eclose have difficulty in fluid-filling of the wings. These studies show that this member of the calcium channel alpha1 subunit gene family plays a nonredundant, vital role in larvae and adults.

Alleles↗

Functions of the N- and C-terminal domains of human RAP74 in transcriptional initiation, elongation, and recycling of RNA polymerase II.

Transcription factor IIF (TFIIF) cooperates with RNA polymerase II (pol II) during multiple stages of the transcription cycle including preinitiation complex assembly, initiation, elongation, and possibly termination and recycling. Human TFIIF appears to be an alpha2beta2 heterotetramer of RNA polymerase II-associating protein 74- and 30-kDa subunits (RAP74 and RAP30). From inspection of its 517-amino-acid (aa) sequence, the RAP74 subunit appears to comprise separate N- and C-terminal domains connected by a flexible loop. In this study, we present functional data that strongly support this model for RAP74 architecture and further show that the N- and C-terminal domains and the central loop of RAP74 have distinct roles during separate phases of the transcription cycle. The N-terminal domain of RAP74 (minimally aa 1 to 172) is sufficient to deliver pol II into a complex formed on the adenovirus major late promoter with the TATA-binding protein, TFIIB, and RAP30. A more complete N-terminal domain fragment (aa 1 to 217) strongly stimulates both accurate initiation and elongation by pol II. The region of RAP74 between aa 172 and 205 and a subregion between aa 170 and 178 are critical for both accurate initiation and elongation, and mutations in these regions have similar effects on initiation and elongation. Based on these observations, RAP74 appears to have similar functions in initiation and elongation. The central region and the C-terminal domain of RAP74 do not contribute strongly to single-round accurate initiation or elongation stimulation but do stimulate multiple-round transcription in an extract system.

Amino Acid Sequence↗

[Effect of various levels of vitamin K intake on bone metabolism of rat].

OBJECTIVE: To explore the effect of various levels of vitamin K intake on bone development. METHODS: Forty weanling Wistar male rats were divided into four groups. In one group, 1% sulfadiazine was added to regular diet (vitamin K 50 micrograms/kg) to induce vitamin K deficiency. In the other three groups, the vitamin K levels in diets were 50 micrograms/kg, 300 micrograms/kg, 2,550 micrograms/kg respectively. Twelve weeks later, the rats were killed and the effects of the different levels of vitamin K intake on bone development were evaluated by the parameters of calcium metabolism, bone metabolic biochemistry, and bone mineral density (BMD). RESULTS: Vitamin K did not affect the intestinal absorption of calcium. Vitamin K deficiency led to the high levels of urinary calcium and hydroxyapatite excretion, suggesting an increase of bone absorption. Different levels of dietary vitamin K significantly affect circulating osteocalcin and OCbound content. The level of serum2 osteocalcin, OCbound and BMD elevated with the increase of dietary VK levels, whereas the parameters of PTH (thrombo plastin time) were not different between all groups. CONCLUSION: Vitamin K can enhance the bone development of rat. The rat vitamin K requirement may be higher than that of the current recommendation (50 micrograms/kg).

Animals↗

Functional expression and characterization of skeletal muscle dihydropyridine receptors in Xenopus oocytes.

Dihydropyridine receptors in vertebrate skeletal muscle serve a dual role: as voltage sensors for excitation-contraction coupling and as voltage-activated calcium channels. Although they were the first of six classes of calcium channels to be cloned, skeletal muscle dihydropyridine receptors remain the only ones not functionally expressed as calcium channels in Xenopus oocytes, leading to the hypothesis that an interacting component is missing. Using beta1b, an isoform previously found in brain, we have for the first time reconstituted skeletal muscle calcium channel function in Xenopus oocytes. We show that this beta subunit is necessary for functional expression and that the alpha2delta subunit significantly enhances the expressed current. The majority of the alpha1 subunit in skeletal muscle is a truncated form. Here we show that both the full-length and truncated forms produce functional calcium channels in Xenopus oocytes, but the truncated form gives significantly larger currents. In addition, we show that the beta1b transcript is expressed in rat skeletal muscle, although at a much lower level than the abundant beta1a isoform.

Animals↗

Effect of amlodipine on the growth of vascular smooth muscle cells of spontaneously hypertensive rats.

The effect of anti-hypertensive drug amlodipine on regression of cardiovascular hypertrophy due to hypertension was studied by using cultured smooth muscle cells derived from arteries of spontaneously hypertensive rats (SHR) and measuring [3H]-TdR and [3H]-Leucine binding. 48 h after adding amlodipine, [3H]-TdR binding in arterial smooth muscle cells from SHR in vitro was reduced by 50.5% and [3H]-Leucine binding was reduced by 56.2% as compared with neuropeptide Y (NPY)-treated group. However, there was no significant change in cell number. The results showed that amlodipine could effectively inhibit increase of DNA and protein synthesis of vascular smooth muscle cell (VSMC) due to NPY. It indicates that amlodipine is of great significance on regression of genesis and development of cardiovascular hypertrophy due to hypertension.

Amlodipine↗

Effects of albumin coating of knitted Dacron grafts on transinterstitial blood loss and tissue ingrowth and incorporation.

Transinterstitial blood loss at implantation and the degree of graft incorporation and inner capsule thickening was compared in serial explants of albumin-coated Dacron versus blood preclotted Dacron grafts in the canine thoracoabdominal aortic position (8 mm internal diameter x 30 cm length). The coated grafts were Bard DeBakey Vasculour II knitted Dacron prostheses impregnated with carbodiimide-cross-linked human albumin. Control grafts were otherwise identical and preclotted with the recipients' whole blood before heparinization during surgery. Transinterstitial blood loss after establishing flow was measured by weighing sponges wrapped around the grafts. Albumin pretreatment resulted in significantly less median blood loss (5.1 g versus 11 g, P=0.04; Mann-Whitney rank sum test). Grafts were explanted at 1 week, 4 weeks, 10 weeks, and 20 weeks. Patency was 100% in both groups. Graft incorporation at explantation was graded by the surgeon as: 1 = none, 2 = minimal, 3 = moderate, or 4 = extensive. No significant differences were noted at any time period. Inner capsule thickness measurements were made every 2.5 mm along the length of all explants. Grafts explanted at 1 week displayed no inner capsules. By 20 weeks, median inner capsule thickness was significantly less in albumin-coated grafts (190 microm versus 235 microm; P<0.0001). These inner capsules in both groups formed as islands, containing abundant myofibroblasts and collagen, covered by endothelial cells and surrounded by residual fibrin coagula. In conclusion, albumin-coated knitted Dacron grafts displayed less transinterstitial blood loss at implantation, and qualitatively similar incorporation, but significantly thinner inner capsules at 20 weeks.

Animals↗

[The change of serotonin of venous blood in pregnancy induced hypertension].

OBJECTIVE: To provide evidence to the hypothesis that the involvement of serotonin (5-HT) takes part in the pathogenesis of pregnancy induced hypertension (PIH). METHOD: 31 pregnant women with PIH and 20 normotensive pregnant women were studied in the present study. Venous plasma 5-HT and platelet 5-HT from all subjects were measured with spectrometry. RESULTS: The mean platelet 5-HT level in PIH group (24.69 +/- 17.79 micrograms/L) was less than that in normotensive group (91.06 +/- 47.77 micrograms/L) (P < 0.01). The mean plasma 5-HT level in PIH group (48.65 +/- 19.29 micrograms/L) was significantly higher than that in normotensive group (35.39 +/- 12.88 micrograms/L) (P < 0.01). CONCLUSION: Plasma levels of 5-HT in PIH were elevated significantly, this might be largely caused by the increased release of 5-HT by platelet. The results show that elevated level of 5-HT in patients with PIH might take part in the pathogenesis of PIH.

Adult↗

[Immunohistochemical locating and quantitative studies of 5-hydroxytryptamine and 5-hydroxytryptamine receptor in placental villi of patients with pregnancy induced hypertension].

OBJECTIVE: To supply evidence of relationship between pregnancy induced hypertension (PIH) and 5-hydroxytryptamine (5-HT) and 5-hydroxytryptamine receptor (5-HTR) synthesized by placental villi. METHODS: Five normal placentas and 15 placentas of PIH were used for studying localization and quantification of 5-HT and 5-HTR in placental villi by using immunohistochemical and in situ quantitative methods. RESULTS: Syncytiotrophoblasts and capillary endothelium in placental villi of control group showed 5-HT and 5-HTR positive immunoreaction, but in placental villi of group with PIH, syncytiotrophoblasts appeared 5-HT and 5-HTR weak positive immunoreaction and capillary endothelium appeared 5-HT and 5-HTR weak positive or negative immunoreaction. The results of in situ quantitation showed that the content of 5-HT and 5-HTR in placental villus syncytiotrophoblasts of group with PIH were significantly lower than those in the control group. CONCLUSIONS: The content of 5-HT and its receptor in placenta may be related to PIH.

Adult↗

Mechanism of cholesterol reduction to coprostanol by Eubacterium coprostanoligenes ATCC 51222.

The mechanism of reduction of cholesterol to coprostanol by Eubacterium coprostanoligenes ATCC 51222 was studied by incubating the bacterium with a mixture of alpha- and beta-isomers of [4-3H,4-14C]cholesterol. Coprostanol, isolated after incubation of [4-3H,4-14C]cholesterol in a growth medium under anaerobic conditions, retained 97% of the tritium originally present in cholesterol. The majority of this tritium (64%), however, was in the C-6 position in coprostanol, which showed that the conversion of cholesterol into coprostanol by E. coprostanoligenes involved the intermediate formation of 4-cholesten-3-one followed by the reduction of the latter to coprostanol. In resting cell assays in which washed bacterial cells were incubated with micellar cholesterol in phosphate buffer at 37 degrees C, both 4-cholesten-3-one and coprostanone were produced in addition to coprostanol. Furthermore, 5-cholesten-3-one, 4-cholesten-3-one, and coprostanone were converted efficiently to coprostanol by E. coprostanoligenes. These results support the hypothesis that the major pathway for reduction of cholesterol by E. coprostanoligenes involves the intermediate formation of 4-cholesten-3-one followed by reduction of the latter to coprostanol through coprostanone as an intermediate.

Carbon Isotopes↗

Biointeractive polymers and tissue engineered blood vessels.

The regulation of endothelial cell (EC) and smooth muscle cell (SMC) proliferation following vascular interventions is critical to clinical efficacy. Our laboratory has developed a method of impregnating biomaterials with suspensions containing bioactive proteins resulting in the capability of differentially modulating EC and SMC growth in vitro and in vivo following implantation. We have previously reported that 60 mu internodal distance ePTFE grafts impregnated with fibrin glue (FG) containing FGF-1 and heparin develop confluent endothelialization with transiently increased EC and SMC proliferation after 4 weeks in dogs. Thoraco-abdominal implants after 20 weeks were developed significantly thicker (139 mu) inner capsules in response to the FGF. To minimize SMC proliferation we studied the effects of FGF-1, heparin, and thrombin concentrations on SMC growth in vitro. FG caused a 182% increase (P < 0.001) in DNA synthesis. Heparin within FG diminished this effect in a dose-dependant manner, with complete inhibition of FG-induced growth at 500 U ml-1 (versus FG alone, P < 0.001). FGF-1 within FG without heparin had no effect, but together, FGF-1 caused a dose-dependant growth increase while increasing heparin concentrations initially increased and then decreased proliferation. FGF-1 and heparin in the medium of quiescent SMCs had similar effects. Only thrombin concentrations > 3.2 U ml-1 stimulated SMC growth and this stimulation was blocked by heparin. A synergism between FGF and heparin on EC proliferation was also found but without EC growth inhibition in response to higher concentrations of heparin. It is thus possible to modulate the relative proliferative activity of ECs versus SMCs by altering the FGF:heparin ratio. This same system may be useful with other proteins to induce other local affects by the applied protein or systemic affects following release of that protein.

Animals↗

[Cloning and sequence analysis of the gene encoding the second region of MSA1 from three Chinese isolates of Plasmodium falciparum].

Gene encoding the second region of merozoite surface antigen (MSA1-R2) of three Chinese isolates of Plasmodium falciparum from Hainan, Yunnan and Anhui were amplified by PCR, cloned into M13 and sequenced. Comparison among MSA1-R2 sequences from different isolates of P. falciparum shows that there is no difference among the three Chinese isolates. The sequence of MSA1-R2 of the Chinese isolates are highly homologous with that of WELLCOME from West Africa. The sequence flanking the upstream of the tandem repeats is exactly the same as that of MAD-20, but sequence downstream the repeats exhibit the characteristics of K1 allele, indicating that intragenic recombination of the two MSA1 allelic families exists in the Chinese isolates.

Amino Acid Sequence↗

[Immunogenicity of recombinant S. typhimurium ex. pressing a hybrid antigen of Plasmodium falciparum].

We have expressed a 74-peptide hybrid Plasmodium falciparum antigen as fusion protein in attenuated Salmonella typhimurium SL3261. Live organisms were orally immunized Rabbits with a dose of 2 x 10(9)cfu. Specific anti-serum were detected by ELISA after immunization. Obvious Delayed type hypersensitivity (DTH) could be induced by PfAg and GZ-C antigen. The recombinant vaccine had no evident side-effects to the hosts. Our studies indicate that attenuated Salmonella typhimurium SL3261 can express synthetic P. falciparum antigen with several epitopes and live organisms can activate special cell-mediated immunity and humoral immunity.

Animals↗

In vivo platelet deposition on polytetrafluoroethylene coated with fibrin glue containing fibroblast growth factor 1 and heparin in a canine model.

BACKGROUND: We previously reported that the coating of expanded polytetrafluoroethylene (ePTFE) with fibrin glue containing fibroblast growth factor 1 (FGF-1) and heparin accelerates endothelial coverage of grafts implanted into animals. We report here the effect of this surface modification on early platelet deposition. MATERIALS AND METHODS: Nine dogs received 7-cm ePTFE grafts, 60-microns internodal distance, 4-mm internal diameter, as bilateral aortoiliac implants, one coated (luminal cross section and abluminal surface) with fibrin glue (fibrinogen 32.1 mg/mliters, thrombin 0.32 U/mliters) containing FGF-1 (11 ng/mliters and heparin (250 U/mliters), the other uncoated. After 5, 30, or 120 minutes of circulation with blood containing autologous platelets radiolabelled with indium 111, gamma emissions were quantitated on explants and correlated to surface areas measured by computerized planimetry. RESULTS: Both global and segmental comparisons showed significantly (P < 0.05, Student's t-test) less platelet deposition on coated than on uncoated grafts after 120 minutes of circulation, but no difference at 5 and 30 minutes. CONCLUSIONS: In this model, ePTFE coating with fibrin glue containing FGF-1 and heparin shows no adverse effect on early platelet deposition.

Animals↗

Selective stimulation of endothelial cell proliferation with inhibition of smooth muscle cell proliferation by fibroblast growth factor-1 plus heparin delivered from fibrin glue suspensions.

BACKGROUND: Pretreatment of expanded polytetrafluoroethylene grafts with fibrin glue (FG) containing fibroblast growth factor-1 (FGF-1) (10 ng/ml) and heparin (50 units/ml) has been shown to induce a transmural angiogenesis with proliferation of both endothelial cells (ECs) and smooth muscle cells (SMCs) in dogs. To induce EC without SMC proliferation, we studied the effects of different FGF-1:heparin ratios within FG in vitro. METHODS: First passage human umbilical vein ECs (factor VIII+) or primary canine carotid artery SMCs (alpha-actin +) were seeded onto 96-well plates coated with FG containing 10 ng/ml FGF-1 and 0, 5, 50, or 500 units/ml heparin. Control wells were coated with FG without FGF-1 or heparin. Cells were fed standard growth medium without soluble FGF-1 or heparin. Tritiated thymidine (1 microCi/well) was added after 1, 2, or 3 days, and proliferation was assayed by scintillation counting 48 hours later. RESULTS: For both ECs and SMCs, proliferation on FG containing FGF-1 but no heparin was not different from control. EC proliferation on FG containing FGF-1 was significantly increased by addition of 5, 50, or 500 units/ml heparin (+68%, +99%, and +106%, respectively; p (0.0001 for all), reflecting the synergism of FGF-1 by heparin. SMC proliferation was also significantly increased by the addition of 5 or 50 units/ml heparin (+85% and +66%, respectively; p (0.0001 for both). However, SMC proliferation with 500 units/ml heparin was significantly decreased from control (-12%; p = 0.014), reflecting heparin's SMC growth inhibitory activity. CONCLUSIONS: FG containing 10 ng/ml FGF-1 and 500 units/ml heparin stimulates EC proliferation while inhibiting SMC proliferation in vitro. Application of this modified FG to vascular grafts or to arteries after direct or transcutaneous interventions may promote endothelialization without intimal hyperplasia.

Cell Division↗