Biomedical subjects

L Nie

Publications and source records attributed to L Nie.

At least 55 records · Page 3Linked to original sources

PubMed1999

[Effect of promoter-PTH4 on Streptomyces sporulation].

233 bp of promoter--PTH4 was subcloned into Streptomyces plasmid pIJ4083. pIJ4470 was obtained after recombinant plasmid was introduced into Streptomyces coelicolor J1501. Both Streptomyces coelicolor J1501/pIJ4083 and J1501/pIJ4470 were grown on minimal medium (MM) agar containing mannitol and 10 micrograms/mL thiostrepton for 6 days, and then ultrathin sections of colonies were prepared and stained with silver proteinate to observe glycogen biosynthesis in cell by the electron microscope. Result showed that the glycogen was produced by J1501/pIJ4083, whereas little glycogen was produced by J1501/pIJ4471. The dark color spores were not observed in J1501/pIJ4470 with lead stain in contrast with dark color spores which were produced by J1501/pIJ4083, indicating that promoter PTH4 may play an important role in physiological and morphological differentiation of Streptomyces.

Gene Expression Regulation, Bacterial↗

PubMed1998

Role of the transmembrane sequence of spleen focus-forming virus gp55 in erythroleukemogenesis.

The membrane glycoprotein encoded by the env gene of either the polycythemia- or anemia-inducing spleen focus-forming virus (SFFVp or SFFVa, respectively) is responsible for the induction of erythroleukemia in mice. It has been shown that the SFFVp glycoprotein, gp55, interacts with the erythropoietin receptor (EPO-R) and promotes EPO-independent proliferation of an EPO-R-expressing hematopoietic cell line, Ba/F3 (Li et al., Nature 343:762, 1990). We show here that when residues within the transmembrane (TM) sequence of an SFFVp gp55 are altered based on the sequences of the anemia-inducing gp55s by a methionine-to-isoleucine (M-I) substitution, a di-leucine deletion (dLL), or both, the resulting mutants display an attenuated phenotype that resembles an SFFVa: they induce milder erythroproliferative disease without polycythemia in vivo and are unable to promote EPO-independent cell proliferation in vitro. The dLL mutation directly interferes with EPO-R binding by decreasing the affinity of gp55 for the receptor. On the other hand, the M-I mutation hampers the full mitogenic activation of EPO-R while having no effect on receptor binding and asserts a dominant negative effect over the wild-type SFFVp gp55. Two other sequence changes within the TM sequence did not affect the biological activities of the SFFVp gp55. These results indicate that the TM sequence of the SFFV env glycoprotein plays a prominent role in SFFV-induced erythroleukemogenesis through its influence on the mitogenic activation of EPO-R.

3T3 Cells↗

PubMed1998

Use of a bioabsorbable polymer for the delivery of ofloxacin during experimental osteomyelitis treatment.

This study was performed to investigate the potential utility of ofloxacin-impregnated bioabsorbable polymers for osteomyelitis therapy. Pseudomonas aeruginosa osteomyelitis was induced in 48 New Zealand White rabbits. Four weeks after infection, the animals were randomized to one of four treatment groups: drug-free polymer, ofloxacin polymer, systemic ofloxacin, or ofloxacin polymer plus systemic ofloxacin. Twenty-eight days later, radiographs were taken of the affected area, the animals were killed, and bone was obtained for histologic evaluation, culture, and determination of ofloxacin concentrations. The percentage of sterile bone cultures was 33, 83, 75, and 91 for the groups treated with drug-free polymer, ofloxacin polymer, systemic ofloxacin, and ofloxacin polymer plus systemic ofloxacin, respectively. When compared with the drug-free polymer, both the ofloxacin polymer and the ofloxacin polymer plus systemic ofloxacin significantly improved the rate of sterilization. The mean concentrations of the drug in bone for the groups treated with ofloxacin polymer, systemic ofloxacin, and ofloxacin polymer plus systemic ofloxacin were 34.9 (range: 2-160), 1.9 (range: 0.8-3), and 26.0 microg/g (range: 9-100 microg/g), respectively. These data suggest that the DL-lactide:glycolide polymer studied is a suitable vehicle for the delivery of high local concentrations of ofloxacin and that these concentrations result in eradication of the bacterial pathogen in this rabbit model.

Animals↗

PubMed1998

Prophylaxis of acute osteomyelitis with absorbable ofloxacin-impregnated beads.

We investigated the use of an ofloxacin-impregnated bioabsorbable composite for the prevention of acute Staphylococcus aureus osteomyelitis. New Zealand White rabbits were anesthetized, the femur was exposed, and a cortical hole was drilled. Animals were randomly given drug-free composites or ofloxacin-impregnated composites; the composites were placed at the site of injury, and the incision was closed. One hour later, all animals were intravenously inoculated with 5 x 10(4) CFU of S. aureus and observed for 28 days. Bone culture data revealed that S. aureus was isolated from 3 of 12 rabbits in the ofloxacin composite group and 9 of 11 animals in the control group (P = 0.02). Radiographic evaluation revealed that the drug-free group had a significantly (P = 0.01) greater degree of radiographic evidence of infection than the group given ofloxacin composites. Although a limited number of histologic samples were available, these data also paralleled the radiographic and culture data. This study demonstrates the effectiveness of the implantable ofloxacin bioabsorbable composites to prevent the development of acute osteomyelitis.

Absorption↗

PubMed1998

Activation of calcium-permeable cation channel by insulin in Chinese hamster ovary cells expressing human insulin receptors.

The present study was conducted to examine the ability of insulin receptor to activate the calcium signaling system in Chinese hamster ovary (CHO) cells expressing human insulin receptor (CHO-IR cells). In these cells, insulin evoked the elevation of cytoplasmic free calcium concentration, [Ca2+]c, measured by using fura-2. Insulin-induced increase in [Ca2+]c was blocked by reducing the extracellular calcium concentration to 1 microM or by adding nickel chloride, an inorganic inhibitor of calcium entry. Insulin did not elevate [Ca2+]c in parental CHO cells or in CHO cells expressing mutant insulin receptor lacking an ATP-binding site. When the transmembrane calcium current was measured by perforated whole-cell patch clamp, adding insulin to the bath solution markedly augmented the inward calcium current. In a cell-attached patch, a single channel activity appeared when insulin was included in the pipette. In contrast, insulin added outside the patch was ineffective. The current/voltage relationship demonstrated that insulin activated a voltage-independent calcium-permeable cation channel with a single-channel conductance of 10 pS. Exposing CHO-IR cells to pertussis toxin abolished the subsequent insulin effect on [Ca2+]c and activation of the calcium-permeable channel. Mastoparan activated the 10-pS calcium-permeable cation channel. In an inside-out patch, insulin activated the calcium-permeable channel when the bath solution contained both GTP and ATP. Nonhydrolyzable ATP could substitute for ATP. These results indicate that in CHO-IR cells, insulin elevates [Ca2+]c by activating the 10-pS calcium-permeable cation channel. Activation by the insulin receptor involves pertussis toxin-sensitive G protein.

Adenosine Triphosphate↗

PubMed1998

[The combined effects of MCYST and water organic pollutants on the co-induction of SHE cell transformation and expression of Ras P21 protein].

The authors studied the combined carcinogenic activities of MCYST and organic pollutants in water of Dianshan Lake, using in vitro Syrian hamster embryo (SHE) cell transformation and immunohistochemical assays. The results showed that MCYST alone could not induce SHE cell transformation, but it increased the effects of low dose organic pollutants in cell transformation in a dose-dependent manner. In the cells isolated from type II transformed foci, ras P21 protein showed a positive immunohistochemical staining. It suggests MCYST may be a tumor promoter, which has a synergistic carcinogenic activity organic pollutants in water and the activation of ras oncogene may be one of the factors inducing malignant transformation.

Animals↗

PubMed1998

Body mass index and hepatic lipase gene (LIPC) polymorphism jointly influence postheparin plasma hepatic lipase activity.

The -514 polymorphism in the hepatic lipase gene (LIPC) is associated with decreased hepatic lipase activity. In the present study, the interaction between body mass index (BMI), the -514 polymorphism, and hepatic lipase activity was examined in 118 white men and in 51 African American men. BMI was significantly positively correlated with hepatic lipase activity in both populations. BMI was similar in men with genetic differences in hepatic lipase activity, indicating that high hepatic lipase activity did not cause increased BMI. The data therefore suggest that high BMI leads to increased hepatic lipase activity. The actions of BMI and the -514 polymorphism on hepatic lipase activity appear to be additive and independent, rather than synergistic. This finding indicates that hepatic lipase activity is a multifactorial trait, determined in part by polymorphism within the LIPC gene as well as by factors that influence BMI.

Adolescent↗

PubMed1998

Three polymorphisms associated with low hepatic lipase activity are common in African Americans.

We have shown previously that a hepatic lipase allele (designated -514T) is common among African Americans and contributes to low hepatic lipase activity in this population. To identify other hepatic lipase alleles associated with low hepatic lipase activity in this population, the coding region and intron-exon boundaries of the hepatic lipase gene were sequenced in 20 African American men with low hepatic lipase activity. Two polymorphisms (N193S and L334F) were associated with low post-heparin plasma hepatic lipase activity and were much more common in African Americans than in whites. This finding, together with our previous data on the -514T allele, indicates that at least three different hepatic lipase polymorphisms associated with low hepatic lipase activity are common among African Americans. Analysis of hepatic lipase haplotypes revealed that 97% of African Americans have at least one hepatic lipase allele that is associated with low hepatic lipase activity.

Adult↗

PubMed1997

Activation of a calcium-permeable cation channel CD20 expressed in Balb/c 3T3 cells by insulin-like growth factor-I.

CD20 functions as a calcium-permeable cation channel. When expressed in Balb/c 3T3 cells, CD20 accelerates the G1 progression induced by insulin-like growth factor-I (IGF-I). To further characterize how CD20 modulates the action of IGF-I, we investigated whether the activity of CD20 channel was affected by IGF-I. In quiescent cells expressing CD20, IGF-I increased cytoplasmic free calcium concentration, [Ca2+]c, which was reversed by the removal of extracellular calcium. In contrast, IGF-I did not increase [Ca2+]c in cells that did not express CD20. In perforated patch clamp recordings, addition of IGF-I to the bath solution augmented the Ca2+ permeability, which was reversed by anti-CD20 antibody. In cell-attached patch, calcium-permeable channel activity with unitary conductance of 7 picosiemens was detected, which was abolished by anti-CD20 antibody. The single channel activities were markedly enhanced when IGF-I was included in the pipette solution, whereas IGF-I added to the bath solution was ineffective. When cells were first exposed to pertussis toxin, activation of the channel by IGF-I was blocked. Transfection of cDNA for Gip2, a constitutive active form of alphai2, activated the CD20 channel. These results indicate that the CD20 channel is regulated by the IGF-I receptor by a mechanism involving pertussis toxin-sensitive G protein.

3T3 Cells↗

PubMed1997

Inhibition of proliferation of MCF-7 breast cancer cells by a blocker of Ca(2+)-permeable channel.

In MCF-7 breast cancer cells, insulin-like growth factor-1 (IGF-1) increased the calcium-permeability of the cells by activating a voltage-independent calcium-permeable channel. IGF-1 also induced oscillatory elevation of cytoplasmic free calcium concentration in these cells. An anti-allergic compound, tranilast, reduced the calcium-permeability augmented by IGF-1 in a dose-dependent manner and blocked the oscillatory elevation of cytoplasmic free calcium concentration. Tranilast did not affect early intracellular signals activated by IGF-1, including receptor autophosphorylation, activations of Ras, mitogen-activated protein kinase and phosphatidylinositol 3-kinase. Tranilast inhibited increases in [3H]-thymidine incorporation, DNA content and cell number induced by IGF-1. The ID50 for [3H]-thymidine incorporation and DNA content were about 10 microM. The inhibitory effect of tranilast was reversible, and cell viability was not affected. Treatment with tranilast increased the number of cells in the G1 phase suggesting that this compound induced G1 arrest. Tranilast also reduced the phosphorylation of the retinoblastoma protein. These results indicate that tranilast inhibits the IGF-1-induced cell growth in MCF-7 cells by blocking calcium entry.

Breast Neoplasms↗

PubMed1997

Detection and analysis of the growth characteristics of Proteus vulgaris with a bulk acoustic wave ammonia sensor.

A bulk acoustic wave ammonia sensor to monitor the growth of Proteus vulgaris is described. The corresponding bacterial growth response equation was derived based on the basic growth characteristics of P. vulgaris, and was verified experimentally. The application of the equation to the estimation of kinetic parameters (such as the asymtote, the maximum specific growth rate and the lag time) was accomplished successfully. The generation time is 33.3 min at 37 degrees C. The influence of temperature on bacterial growth was also investigated and the activation energy of P. vulgaris growth was estimated. Furthermore, the initial bacterial concentrations were determined based on the linear relationship between the inflection time, ti, and the logarithm of the initial bacterial concentration in the range 10(2)-10(7) (cells ml-1). The results were in good agreement with those from the pour plate count method and the precision was better than that of the frequency detection time method.

Acoustics↗

PubMed1996

Kinetic assay of antitrypsin in human serum by a surface acoustic wave(SAW)-impedance sensor.

Antitrypsin in human serum was determined by using both the SAW-impedance sensor system and spectrophotometry, indicating that the mean value for women was significantly higher than the mean value for men; the value for acute pancreasis patients is about 2-folds of the normal values, and there is no significant difference between the acute pancreasis patients and the pancreatic cancer patients.

Journal Article↗

PubMed1996

A new urea sensor based on combining the surface acoustic wave device with urease extracted from green soya bean and its application--determination of urea in human urine.

The urea sensor was prepared by combining a surface acoustic wave (SAW) device, in which a SAW resonator operating at 61 MHz and a pair of parallel electrodes were used in series, with urease extracted from green soya bean. The Michaelis constant and maximum reaction rate of the urease were estimated as 2.14 mM and 27.18 kHz min-1, respectively, at pH 7.0 and 25.0 degrees C. Influences of pH, temperature and effectors on the response properties of the SAW urea sensor were investigated. Recovery of the sensor ranged from 95 to 105% and the detection limit of urea was 1.0 micrograms ml-1 (1.7 x 10(-5) M). The proposed sensor has been successfully applied to the rapid determination of urea in human urine samples. The results are consistent with the reported values and also support the clinical diagnosis.

Acoustics↗

PubMed1996

Behaviour of surface acoustic wave interdigitated array electrode sensor in non-aqueous solution and determination of blood plasma recalcification time.

A novel SAW-IDA sensor system was constructed for the first time by connecting the IDA electrodes in series with a SAW resonator. The frequency characteristics of the SAW-IDA sensor in non-aqueous solution were investigated. The effects of the parallel capacitance and cell constant were studied, and were calculated with the circuit network theory. These calculations provide guiding rules for design of the SAW-IDA sensor system. The SAW-IDA sensor was applied to determination of recalcification time and activated partial thromboplastin time of blood plasma. The sensor offers a new and effective way of studying clinical and laboratory haemostasis.

Acoustic Stimulation↗

PubMed1996

Distinct local anesthetic affinities in Na+ channel subtypes.

Lidocaine is a widely used local anesthetic and antiarrhythmic drug that is believed to exert its clinically important action by blocking voltage-gated Na+ channels. Studies of Na+ channels from different species and tissues and the complexity of the drug-channel interaction create difficulty in understanding whether there are Na+ channel isoform specific differences in the affinity for lidocaine. Clinical usage suggests that lidocaine selectively targets cardiac Na+ channels because it is effective for the treatment of arrhythmias with few side effects on muscle or neuronal channels except at higher concentrations. One possibility for this selectivity is an intrinsically higher drug-binding affinity of the cardiac isoform. Alternatively, lidocaine may appear cardioselective because of preferential interactions with the inactivated state of the Na+ channel, which is occupied much longer in cardiac cells. Recombinant skeletal muscle (hSkM1) and cardiac sodium channels (hH1) were studied under identical conditions, with a whole-cell voltage clamp used to distinguish the mechanisms of lidocaine block. Tonic block at high concentrations of lidocaine (0.1 mM) was greater in hH1 than in hSkM1. This was also true for use-dependent block, for which 25-microM lidocaine produced an inhibition in hH1 equivalent to 0.1 mM in the skeletal muscle isoform. Pulse protocols optimized to explore inactivated-state block revealed that hSkM1 was five to eight times less sensitive to block by lidocaine than was hH1. The results also indicate that relatively more open-state block occurs in hSkM1. Thus, the cardiac sodium channel is intrinsically more sensitive to inhibition by lidocaine.

Anesthetics, Local↗

PubMed1996

[Changes in blood gas in burned patients in Xining].

The changes in arterial blood gas were observed and analysed in twenty-seven burned patients at high altitude. It was showed that a serious hypoxemia existed in burn patients, both in survivors and dead cases, and that hypoxemia occurred earlier in the cases with inhalation injury or dead cases than without inhalation injury. These results suggest that patients with inhalation injury should undergo tracheostomy as early as possible to ensure oxygen supply, and patients without inhalation injury should be give oxygen therapy routinely. ARDS occurred in all inhalation injury cases, and high frequency jet ventilation was effective in improving hypoxemia in such cases.

Adolescent↗

PubMed1996

Blockade of DNA synthesis induced by platelet-derived growth factor by tranilast, an inhibitor of calcium entry, in vascular smooth muscle cells.

The present study was conducted to establish a pharmacological method of controlling growth of vascular smooth muscle cells (VSMC) by blocking calcium entry. In cultured rat VSMC, 1 nM platelet-derived growth factor (PDGF) induced a biphasic elevation of cytoplasmic free calcium concentration, ([Ca2+]c). The second sustained phase of [Ca2+]c was dependent on extracellular calcium. At lower concentrations, PDGF induced oscillatory changes in [Ca2+]c, and reduction of extracellular calcium attenuated the oscillation. An antiallergic compound, tranilast, abolished the sustained phase of [Ca2+]c induced by 1 nM PDGF. Tranilast also inhibited the oscillatory changes in [Ca2+]c induced by 200 pM PDGF. In addition, PDGF-induced calcium influx in the late G1 phase, as assessed by measuring the initial uptake of 45Ca, was inhibited by tranilast in a concentration-dependent manner. Tranilast also inhibited PDGF-augmented DNA synthesis; the ID50 for the inhibition of DNA synthesis was nearly identical to that for calcium influx. Although tranilast blocked PDGF-induced calcium entry, it did not affect PDGF-mediated autophosphorylation of the PDGF receptor, activation of phosphatidylinositol 3-kinase, activation of Ras or mitogen-activated protein kinase. Similarly, PDGF-induced elevation of diacylglycerol was not affected by tranilast. These results suggest that the antiallergic drug tranilast inhibits PDGF-induced DNA synthesis by blocking PDGF-mediated calcium entry. Tranilast may be of use in controlling PDGF-induced DNA synthesis in VSMC.

Animals↗

PubMed1995

Assay of pancreatic lipase with the surface acoustic wave sensor system.

A new type of surface acoustic wave sensor system for assaying the activity of pancreatic lipase has been proposed. The assay of this enzyme is based on the change in conductance of the solution caused by the release of a fatty acid, using triolein as a substrate. A linear relationship between frequency response and enzyme concentration is obtained. Kinetic parameters of pancreatic lipase, i.e., the Michaelis constant for triolein and the corresponding maximum initial rate, are estimated to be 5.46 mM and 7551.6 Hz/min, respectively. The effects of temperature and pH value are also investigated.

Acoustics↗