Biomedical subjects
Publications and source records attributed to N Liu.
This paper dealed with the effect of media fertilization on the quality of Gastrodia eleta according to the results of tissue culture. The results showed that contents of nutrient elements, gastrodin and free amino acids in G. elata were affected by various nutrients solutions. The result of comprehensive analysis showed that the 1/2 MS medium was the fittest treatment of fertilization.
OBJECTIVE:To study the effect of N(2)O inhalation and propofol intravenous anaesthesia in tooth extraction. METHODS: 40 dental outpatients with the extraction of the mandibular wisdom tooth,ASAI,were randomly allocated to two groups:N(2)O group with the age of 20-36 years old and propofol group with the age of 21-35 years old.All cases undergoing dental extraction were given ultra-light anaesthesia as an adjunct local anaesthesia. RESULTS: The two groups were gained various levels of sedation,there were no significant difference among BP,HR,RR,SPO(2) before,in,and after the operation.Dental treatment could be carried out in cooperating conditin throughout the whole procedure.There was a certain extent of amnesia postoperation,the incidence of postoperative side effects was low,the recovery time was gained within 30 minutes. CONCLUSION: The analgesic effects of N(2)O group were superior to the propofol group.
The cdc25C , cdc2 and cyclin A promoters are controlled by transcriptional repression through two contiguous protein binding sites, termed the CDE and CHR. In the present study we have identified a factor, CDF-1, which interacts with the cdc25C CDE-CHR module. CDF-1 binds to the CDE in the major groove and to the CHR in the minor grove in a cooperative fashion in vitro , in a manner similar to that seen by genomic footprinting. In agreement with in vivo binding data and its putative function as a periodic repressor, DNA binding by CDF-1 in nuclear extracts is down-regulated during cell cycle progression. CDF-1 also binds avidly to the CDE-CHR modules of the cdc2 and cyclin A promoters, but not to the E2F site in the B- myb promoter. Conversely, E2F complexes do not recognize the cdc25C CDE-CHR and CDF-1 is immunologically unrelated to all known E2F and DP family members. This indicates that E2F- and CDF-mediated repression is controlled by different factors acting at different stages during the cell cycle. While E2F-mediated repression seems to be associated with genes that are up-regulated early (around mid G1), such as B- myb , CDE-CHR-controlled genes, such as cdc25C , cdc2 and cyclin A , become derepressed later. Finally, the fractionation of native nuclear extracts on glycerol gradients leads to separation of CDF-1 from both E2F complexes and pocket proteins of the pRb family. This emphasizes the conclusion that CDF-1 is not an E2F family member and points to profound differences in the cell cycle regulation of CDF-1 and E2F.
B- myb and cdc25C exemplify different groups of genes whose transcription is consecutively up-regulated during the cell cycle. Both promoters are controlled by transcriptional repression via modules consisting of an E2F binding site (E2FBS) or the related CDE plus a contiguous CHR co-repressor element. We now show that the B- myb repressor module, which is derepressed early (mid G1), is preferentially recognized by E2F-DP complexes and that a mutation selectively abolishing E2F binding impairs regulation. In contrast, the cdc25C repressor module, which is derepressed late (S/G2), interacts selectively with CDE-CHR binding factor-1 (CDF-1). E2F binding, but not CDF-1 binding, requires specific nucleotides flanking the E2FBS/CDE core, while CDF-1 binding, but not E2F binding, depends on specific nucleotides in the CHR. Swapping these nucleotides between the two promoters profoundly changes protein binding patterns and alters expression kinetics. Thus predominant CDF-1 binding leads to derepression in late S, predominant E2F binding results in up-regulation in late G1, while promoters binding both E2F and CDF-1 with high efficiency show intermediate kinetics. Our results support a model where the differential binding of E2F and CDF-1 repressor complexes contributes to the timing of promoter activity during the cell cycle.
Bovine prochymosin (chymosin) contains three disulfide bonds: Cys45-Cys50, Cys206-Cys210 and Cys250-Cys283 (pepsin numbering). We have demonstrated that Cys250-Cys283 is not intimately involved in the catalytic mechanism of chymosin but is required for the correct refolding of prochymosin. To elucidate the functional implications of another disulfide bond of Cys45-Cys50, these two cysteine residues were replaced separately or simultaneously by site-directed mutagenesis. Like the wild-type prochymosin all the seven mutants generated (C45A, C50A, C45A/C50A, C45D, C50S, C45D/C50S, C45A/C50S) exhibit the activity of autocatalytic activation after refolding, indicating that Cys45-Cys50 is dispensable in prochymosin refolding. Spectroscopic analyses and urea-induced denaturation studies of prochymosin and four mutants tested (C45A, C50A, C45A/C50A, C45D/C50S) show that: (1) they share similar far-UV CD spectra and similar fluorescence emission spectra; (2) mutation results in a perturbance of tryptophan environment and somewhat destabilization of prochymosin conformation. However, quenching studies reveal that the only one tryptophan residue inaccessible to acrylamide is still buried in the mutated molecules. All these results suggest that the overall conformation of prochymosin is maintained after mutation. As for the enzymatic properties of pseudochymosin, the activation product of prochymosin, it has been found that elimination of Cys45-Cys50 causes a marked drop of thermostability and an alteration of substrate specificity.
GH is synthesized at multiple extrapituitary sites suggestive of an autocrine/paracrine mechanism of action. We have investigated a possible autocrine/paracrine mechanism of GH action, compared the cellular response to exogenous versus endogenously produced GH, and determined the nature of the interaction between external stimuli and endogenously produced GH. BRL cells expressing the GH receptor were transiently transfected with expression plasmids containing either the hGH or the bGH gene and the response of the cell was measured by CAT reporter plasmids requiring either STATs 1 and 3 or STAT5 for their response. Transient transfection of the hGH gene resulted in hGH accumulation in the cell and secretion into the media. The functional response through STATs 1 and 3 and STAT5 obtained with endogenously produced hGH was comparable or greater in magnitude to that obtained with the maximal stimulatory dose of exogenous hGH. Similar results were obtained with an expression plasmid containing the bGH gene. Endogenously produced hGH interacted in an additive manner when combined with submaximal doses of both exogenous hGH and serum. Such results were also observed in a more physiologically relevant mammary carcinoma cell line (MCF-7). The nonreceptor-dimerizing hGH antagonist, hGH-G120R, used in cells expressing the homologous receptor extracellular domain was able to only partially inhibit the response of the cell to endogenously produced hGH, in contrast to full inhibition of exogenous hGH. We therefore conclude that GH can function in an autocrine/paracrine manner, additive in effect to external stimuli.
The Chinese hamster ovary (CHO) mutant UV40 cell line is hypersensitive to UV and ionizing radiation, simple alkylating agents, and DNA cross-linking agents. The mutant cells also have a high level of spontaneous chromosomal aberrations and 3-fold elevated sister chromatid exchange. We cloned and sequenced a human cDNA, designated XRCC9, that partially corrected the hypersensitivity of UV40 to mitomycin C, cisplatin, ethyl methanesulfonate, UV, and gamma-radiation. The spontaneous chromosomal aberrations in XRCC9 cDNA transformants were almost fully corrected whereas sister chromatid exchanges were unchanged. The XRCC9 genomic sequence was cloned and mapped to chromosome 9p13. The translated XRCC9 sequence of 622 amino acids has no similarity with known proteins. The 2.5-kb XRCC9 mRNA seen in the parental cells was undetectable in UV40 cells. The mRNA levels in testis were up to 10-fold higher compared with other human tissues and up to 100-fold higher compared with other baboon tissues. XRCC9 is a candidate tumor suppressor gene that might operate in a postreplication repair or a cell cycle checkpoint function.
Structural studies have demonstrated that the extracellular L3 loop of porin constricts the channel and suggest that this loop might be involved in channel selectivity and gating. We previously showed that positively charged polyamines can induce changes in porin gating kinetics by stabilization of closed states. Here we report the effects of the mutation of two different aspartate residues of Escherichia coli OmpC porin on the polyamine sensitivity of the channel. Aspartate 105 or aspartate 118 on the L3 loop was replaced by glutamine by site-directed mutagenesis. The gating activity of the wild-type and mutant channels were studied by patch-clamp of liposomes containing reconstituted outer membrane fractions, in the absence or the presence of either polyamine spermine or cadaverine. Porin channels with a D118Q mutation, at the root of L3, still showed some, albeit milder, sensitivity to polyamine modulation. On the other hand, the D105Q mutation, at the tip of L3, abolished the increase in closing frequency which is typically observed in the presence of polyamines. We conclude that aspartate 105 primarily, but not aspartate 118, plays an important role in mediating the polyamine-induced changes in gating kinetics that result in the inhibition of the OmpC channel.
A 42 residue artificial peptide that binds to the HIV-1 enhancers has been described previously. The specificity of interaction of the peptide with its target DNA sequence has been demonstrated by a variety of techniques. Naturally occurring regulatory proteins frequently bind to DNA as dimers, thereby increasing the strength and specificity of the interaction, the dimer interface often being provided by a leucine zipper type coiled coil. As a suitable binding site for this kind of system is located to the 5' end of the HIV enhancer region, it was decided to design and synthesize a fusion peptide that not only contained the DNA binding sequence of the original 42 residue peptide but also incorporated a leucine zipper based on that of the GCN4 transcriptional activator, that should, therefore, be capable of dimerizing. The resultant peptide, LZ66, has now been shown to be fully active in band shift and in vitro transcription assays and to exhibit about double the inhibitory activity of the parent 42 residue peptide. Preliminary CD measurements revealed that the peptide has a high alpha-helical content and that it adopts a stable conformation down to the low micromolar peptide concentration range. Sedimentation equilibrium studies confirmed that the principles involved in the design of the peptide are valid and that the peptide is indeed dimeric in solution.
To improve our understanding of phenobarbital (PB) mediated induction of CYP6D1 we examined the genetic linkage of PB induction using PB responsive (aabys) and non-responsive (LPR) strains of house flies. PB induction was linked to autosome 2, indicating that CYP6D1 is trans regulated by this PB inducible element. Our results are discussed relative to a previous hypothesis that the same regulatory genes are responsible for both induction and monooxygenase-mediated insecticide resistance.
The effect of ara A (9-beta-D-arabino furanosyladenine) a potent inhibitor of DNA synthesis on the frequencies of chromatid breaks induced by restriction endonucleases (RE) has been investigated in normal human and ataxia telangiectasia (AT) lymphoblastoid cells. PvuII, PstI and BamHI which cause blunt-ended, 3'-overhang and 5'-overhang cohesive-ended DNA double-strand breaks (dsb) respectively, were introduced into two AT cell lines (AT-KM and AT-PA) and a normal human (N-SW) cell line by the use of streptolysin-O poration. Controls were exposed to gamma-irradiation and similarly treated with or without ara A. Both AT cell lines were found to exhibit higher frequencies of chromatid breaks when treated with RE alone as compared with the normal cell line. The pattern of chromatid response to the three RE was shown to be similar in all three cell lines i.e. PvuII was most clastogenic while PstI and BamHI were both less effective at inducing chromosomal aberrations. Incubation of cells with ara A resulted in an increase in frequencies of chromatid breaks in PvuII and PstI treated cells but no increase was observed in BamHI treated cells. Normal cells showed most response to ara A following treatment with PvuII and PstI (enhancement ratios 4.63 and 3.75 respectively) while AT cells were affected by ara A to a lesser extent indicating a reduced expression of damage by ara A in these lines. Since ara A is a potent inhibitor of DNA synthesis, it was concluded from the elevated frequency of chromosomal aberrations in the presence of ara A that rejoining of RE-induced dsb in genomic DNA of human cells involves nucleotide insertion at dsb termini prior to ligation.
The inheritance of cytochrome P450 monooxygenase-mediated (i.e., CYP6D1) resistance in the LPR strain of house fly, Musca domestica L., was investigated by comparing levels of monooxygenase-mediated permethrin resistance and CYP6D1 messenger RNA in 7 populations of house flies. Results indicate that monooxygenase-mediated resistance, and overexpression of CYP6D1 in the LPR strain are controlled by an incompletely recessive trans acting factor(s) on autosome 2 and a nearly completely dominant cis acting factor(s) on autosome 1. The polygenic nature of CYP6D1-mediated resistance and the relative dominance of the resistance factor(s) on autosome 1 suggests this type of resistance could develop relatively quickly.
UNLABELLED: The aim of this study was to assess whether hypoxemia during one-lung ventilation (OLV) can be prevented by inhaled nitric oxide (NO) (Part I) or by its combination with intravenous (IV) almitrine (Part II) in 40 patients undergoing thoracoscopic procedures. In Part I, 20 patients were divided into two groups: one received O2 (Group 1) and one received O2/NO (Group 2). In Part II, 20 patients were divided into two groups: one received O2 (Group 3) and one received O2/NO/almitrine (Group 4). In Groups 2 and 4, NO (20 ppm) was administered during the entire period of OLV, and almitrine was continuously infused (16 microg x kg(-1) x min[-1]) in Group 4. Arterial blood gases were measured during two-lung ventilation with patients in the supine position, after positioning in the lateral decubitus position, and then every 5 min for a 30-min period during OLV. During OLV, Pao2 values decreased similarly in Groups 1 and 2. After 30 min of OLV, the mean Pao2 values in Groups 1 and 2 were 132 +/- 14 mm Hg (mean +/- sem) and 149 +/- 27 mm Hg (not significant [NS]), and the Pao2 value was less than 100 mm Hg in four patients in Group 1 and five patients in Group 2. Pao2 values were greater in Group 4 than in Group 3 after 15 and 30 min of OLV. After 30 min of OLV, the mean Pao2 values were 146 +/- 16 mm Hg in Group 3 and 408 +/- 33 mm Hg in Group 4 (P < 0.001). Pao2 was less than 100 mm Hg during OLV (NS) in four patients in Group 3 and in no patient in Group 4. We conclude that NO inhalation alone has no effect on Pao2 evolution during OLV, although its combination with IV almitrine limits the decrease of Pao2 during OLV. This beneficial effect of NO/almitrine could be attributed to an improvement in ventilation-perfusion relationships. IMPLICATIONS: Decrease in oxygenation during one-lung ventilation is quite common. Our study showed that inhaled nitric oxide alone did not influence Pao2 evolution. We then tried adding intravenous almitrine to nitric oxide with amazingly good results on Pao2. This nonventilatory technique should be of great use during special thoracic acts, such as thoracoscopic procedures.
The objective of the study was to evaluate the level of 8-hydroxydeoxyguanosine (8-OHdG) in DNA of peripheral-blood leukocytes as a biological marker of asbestos exposure and/or its fibrotic effects in an occupational population exposed to asbestos. The setting was a large-scale asbestos plant in China producing brake linings, asbestos rubber, and textile using chrysotile. From a base population of active and retired workers with various levels of cumulative exposure to asbestos and grades of asbestosis, 39 study subjects were randomly selected to reflect incremental grades of asbestosis based on Chinese diagnostic standards. They consisted of 19 "normal" (control) and ten "suspected" and ten "definite" asbestosis-grade subjects, group-matched for age and sex. Leukocytic DNA was extracted from 5-mL samples of peripheral blood and 8-OHdG level measured by high-pressure liquid chromatography. A cumulative asbestos exposure index (CEI) was calculated for each subject as the summed product of duration and level of asbestos exposure per job, incorporating a job-exposure matrix. Geometric mean 8-OHdG levels showed a positive gradient in relation to increasing grades of asbestosis (control: 1.78, suspected: 2.21, definite: 2.58), with a significant difference between the control and definite-asbestosis subgroups (p < 0.05). The 8-OHdG level of the two subgroups combined as one "asbestosis" group was significantly higher than that of the control group (control: 1.78, asbestosis: 2.39, p = 0.01). Further, 8-OHdG levels were moderately correlated with CEIs for all subjects (r = 0.35, p < 0.05) and with grades of asbestosis for all (r = 0.47, p < 0.01) and for male subjects (r = 0.43, p < 0.05). In multiple regression analyses, grade of asbestosis explained 27% of the total variation in 8-OHdG and was a better predictor than CEI or duration of exposure. Thus, the 8-OHdG level in leukocytic DNA is related to grade of asbestosis and to individual cumulative exposure and may serve as a biologic marker reflecting the status of oxidative DNA damage by asbestos.
BACKGROUND: In patients after lung transplantation, dysfunction of pulmonary venous and artery anastomoses leading to reoperation is described. METHODS AND RESULTS: Pulmonary artery and vein anastomoses were evaluated intraoperatively by monoplane transesophageal echocardiography (TEE) in 18 patients undergoing lung transplantation (nine right, five left single lung transplantations, and four bilateral transplantations). All 13 right pulmonary artery anastomoses and all 22 pulmonary vein anastomoses could be visualized by TEE. None of the nine left pulmonary anastomoses could be visualized. Of the 13 right pulmonary anastomoses, 12 were considered normal, their diameter ranging from 1 to 1.7 cm (mean, 1.26 +/- 0.24 cm). A moderate stenosis of one pulmonary artery anastomosis was identified but did not require reoperation. Of the 22 pulmonary vein anastomoses, 16 were considered normal, their diameter being > 0.5 cm and the peak systolic flow velocity < or = 1 m/s at the location of the anastomoses. In five cases, the anastomoses were not considered normal, but reoperation was not indicated. In one case, a severe stenosis of pulmonary vein associated with graft dysfunction led to an early reoperation. CONCLUSION: Intraoperative TEE during lung transplantation contributes to the immediate evaluation of pulmonary vein and right pulmonary artery anastomoses and allows immediate surgical correction. Further investigations are necessary to establish threshold values requiring reoperation.
OBJECTIVE: To observe the protective effect of Huanshaodan (HSD) in mouse brain in a steady ischemia-reperfusion model. METHODS: Step-through experiment, receptor binding test and choline acetyl-transferase (CAT) activity determination was performed. RESULTS: After HSD treatment, mice made much less mistakes in step-through experiment than sham groups. Meanwhile the 3H-MK801 binding of cerebral cortex and hippocampus tissues was significantly decreased. CAT activity of cerebral cortex and hippocampus tissue were increased respectively. CONCLUSION: HSD might antagonize ischemia injury of brain through modulating excessive activation of glutamate N-methyl-D-Aspartic acid receptors.
OBJECTIVE: To explore the modulation of cell proliferation induced by growth factors in cultured human retinal pigment epithelial (RPE) cells. METHODS: A well-cultured system of human fetal RPE cells was established to determine the effects of several growth factors, such as epidermal growth factor (EGF), basic-fibroblast growth factor (b-FGF), insulin-like growth factor (IGF-I), on the cultured human RPE cells by using 3H-thymidine incorporation and cell count. RESULTS: (1) EGF, b-FGF, IGF-I significantly increased the DNA 6.6-12.2 times in human RPE cells in comparison with the control, the rank order of potency of the factors being EGF > b-FGF > IGF-I. (2) When the factors were used in combination, the above factors caused a significant enhancement of 3H-Tdr incorporation by times. CONCLUSION: It is suggested that the synergetic effects of growth factors and their complex interactions play a role in the development of proliferative retinopathy.
The incorporation of [3H]thymidine obviously tends to decrease in the primary cultured hepatocytes of rats when the cells are treated with carbon tetrachloride. Meanwhile the activity of ALT markedly increases in the medium. But these pathologic alterations are inhibited by Salvianic acid B(SA-B) and the effect of 10(-6) mol/L SA-B is the best.