Biomedical subjects
Publications and source records attributed to L Qiu.
The atractylenolide III content in different processed products of Atractylodes macrocephala was determined by HPLC. The result has shown that the content in the processed products appears higher than that in the crude herb, with the yellow-fried product being the highest.
In order to find out the quantitative relationship between physicochemical properties of drugs and their nasal absorption, diltiazem hydrochloride and paracetamol were selected as model drugs and their octanol-water partition coefficient was determined. In situ nasal recirculation method at different pH values was used to estimate the rate constant of nasal drug absorption in rats. Results showed that quantitative relationship existed between partition coefficient and nasal absorption constant, with correlation coefficient being 0.9761(n = 9). Besides methods of partition coefficient determination, the in situ nasal recirculation manipulation was also improved.
Ananalgestic effect of different processed products of Sparganium stoloniferum in mice is observed in hot plate and acetic acid induced writhing test, The experimental results shows that water extract of S. stoloniferum and its processed products can inhibit the pain. The analgestic effect of its processed product with vinegar is the most powerful and lasting.
OBJECTIVE: To study the effect of IFN-alpha on the expressions of perforin and granzymes in IL-2-activated-lymphocytes. METHODS: NK and LAK activities were assayed by 4 hour standard 51Cr release test, the activity of perforin was detected by hemolysis method, expression of granzyme B was measured by ABC immunohistological method, expression of granzyme A was measured by BLT method. RESULTS: IFN-alpha significantly augmented the activities of NK and LAK in IL-2 activated peripheral blood lymphocytes (PBL) after 1 day culture. Perforin activity in lymphocytes was increased after 1 day exposure to IL-2 or IFN-alpha, and was enhanced when exposed to the combination of IL-2 and IFN-alpha. After 3 day culture, the perforin activity remained high in lymphocytes activated by IL-2 alone or in combination with IFN-alpha, while declined to control level in IFN-alpha exposed group. IL-2 and IFN-alpha alone or in combination had no effect on expression of granzyme A and B. CONCLUSION: IFN-alpha enhances the cytotoxicity of lymphocytes activated by IL-2. The mechanism might be that IFN-alpha upregulates the perforin expression.
OBJECTIVE: To investigate the reversal of homoharringtonine (HHT)-resistance by tamoxifen (TAM) or verapamil (VER) alone or in combination. METHODS: The drug-sensitivity test was performed with semisolid agar culture. RESULTS: The cytotoxicity of HHT to K562/S cells was not enhanced by TAM or VER alone or in combination,but HHT resistance in HHT resistant cells (K562/H20) was reversed by VER and TAM at nontoxic doses (4micromol/L or 8micromol/L). The IC50 of K562/H20 for HHT decreased from 446.8 +/- 0.08microg/L to 45.1 +/- 0.02microg/L in the presence of 4micromol/L of VER, to 22.4 +/- 0.03microg/L in 8micromol/L of VER, to 85.1 +/- 0.03microg/L in 4micromol/L of TAM and to 26.4 +/- 0.02microg/L in 8micromol/L of TAM. In the presence of combinations of 2micromol/L VER and 4micromol/L TAM and of 2micromol/L VER and 8micromol/L TAM, IC50 of K562/H20 decreased to 30.4 +/- 0.02microg/L and 4.3 +/- 0.04microg/L, respectively. CONCLUSION: HHT-resistance could be reversed by VER or TAM alone, and the combination of the two drugs showed a synergistic effect.
OBJECTIVE: In order to investigate the life quality of patients with hematological diseases after bone marrow transplantation (BMT). METHODS: Pituitary-thyroid and adrenocortical function were de tected in 29 patients with hematological diseases before and after BMT. RESULTS: All patients had normal thyroid and adrenocortical functions before BMT. As compared with pre-BMT,the median serum TSH levels were significantly increased at 3, 6, 12 (P<0.01), 24-60 (P<0.05) months post-BMT. The adrenocortical function had no change in all patients, and thyroid dysfunction were found in 6 of 24 patients detected at a median time of 6 month post-BMT. Five of the 6 thyroid dysfuntion patients developed hypothyroidism, and one hyperthyroidism. CONCLUSION: There was little effect of conditioning regimen on adrenocortical function, while thyroid dysfunction was common after BMT. Long term follow up on thyroid function in patients after BMT is necessary.
OBJECTIVE: To evaluate the killing effect of IL-2 and IFN-alpha activated bone marrow cells on K562 cells. METHODS: Semi-solid colony assay was used. RESULTS: Bone marrow from leukemia patients in remission was activated in vitro with IL-2 for 3 days. The activated bone marrow (ABM) displayed killing effects of 0.31 approximately 2.30 logs on K562 cells, this killing effects was further increased to 0. 30 approximately > 3.15 logs when IFN-alpha added with IL-2 to the marrow for activation. IL-2 alone or in combination with IFN-alpha showed no inhibition of CFU-GM and K562 cells. Compared with IL-2 or IFN-alpha alone, the combination of the two cytokines could more effectively maintain the killing effect of ABM on leukemic cells. CONCLUSION: IFN-alpha can augment the purging effect of IL-2 ABM and combination of the two cytokines can effectively maintain the cytotoxicity of ABM.
Two hTPO cDNA segments (N-terminal and C-terminal) were amplified from human fetal liver mRNA by using separate reverse-transcription PCR reactions, and cloned into pUC19. Their sequences were identical with that previously reported. Then the full length cDNA of hTPO was obtained from the two cDNA fragments, cloned into the shuttle vector pSVK3 and transiently expressed in COS-7 cells. The activity of the expression product was demonstrated with the stimulation of CFU-Meg.
To examine hammerhead ribozyme structure and vulnerability to cellular nucleases, ribozymes were incubated with soluble extracts from Escherichia coli, and cleavage sites were identified by primer extension analysis. Mapping of endonuclease-sensitive sites revealed that the most sensitive were in the 3'-substrate-binding region of the ribozyme. The catalytic domain was much less susceptible, although some cleavage preference was seen at two positions known to be twisted out of parallel stacking in a ribozyme-substrate analogue complex. Changes in substrate-binding domain nucleotide sequence had no effect on cleavage patterns of catalytic domains. Hammerhead ribozymes, in solution and free from substrate, appear to have structurally independent, asymmetrically arranged domains.
To understand the relationship of inflammatory and cellular stress responses, phospholipase A2 (PLA2) was examined for its role in the first phase of the transcriptional response to cellular stress. Electromobility shift analysis revealed heat shock transcription factor (HSF1)-DNA binding when HeLa S3 and Jurkat cells were exposed to exogenous PLA2. Although PLA2-inducible HSF1-DNA binding was comparable to thermal stress, it did not induce maximal heat shock gene expression. PLA2-induced HSF1 was not hyperphosphorylated relative to the heat-inducible form, thus suggesting that exogenous PLA2 affects the signal for HSF1 multimerization but not its phosphorylation. Because inflammation often involves elevated temperatures, the effect of PLA2 on thermal regulation of HSF1-DNA binding activity was examined. PLA2 exposure altered the thermal threshold for HSF1 activation, and pore-gradient gel analysis indicated that either conformational changes or other modifications of HSF1 are being induced when cells are treated by PLA2, thus creating a synergistic environment for HSF1 activation into its DNA-bound state. Surprisingly, the monocyte-like cell line, U-937, was insensitive to the action of exogenous PLA2. Neither HSF1-DNA binding or lowering of the temperature threshold for HSF1 activation was observed in PLA2-treated U-937 cells. These data suggest that inflammatory mediators such as PLA2 partially affect transcriptional switches mediating thermal stress in some cell types but not others. The purpose of HSF1 activation during inflammation and its differential induction are discussed relative to these observations.
In this paper beta-globin gene mutations were detected in 452 beta-thalassemia carriers from 13462 unselected individuals (6731 pregnant women and their husbands) who were screened for the beta-thalassemia trait in the Guangzhou area of China. The incidence of beta-thalassemia was calculated as 3.36%. This is higher than found in previous studies performed in southern China. Using reverse dot blot analysis, we found 11 types of mutations and identified the mutations in 446 (98.7%) of the 452 cases. Direct sequencing was carried out on the 6 unknown alleles, and a novel amber mutation in a beta 0-thalassemia gene (beta 37TGG --> TAG) was found in one of them. Thus, the prevalence and spectrum of beta-thalassemia mutations were obtained for this region. Twelve couples were detected at risk for thalassemia, and prenatal diagnosis was carried out in 11 of them. This is the largest number of Chinese subjects studied by DNA analysis to date and is the first report on the prospective diagnostic trial for beta-thalassemia in a Chinese population. In addition, we have performed 80 prenatal diagnoses based on screening for beta-thalassemia retrospectively.
Hammerhead ribozymes are small catalytic RNA molecules that can be designed to specifically cleave other RNAs. These ribozymes have exhibited low efficiency when examined inside cells, perhaps in part because of their sensitivity to intracellular RNases. In an effort to better understand intracellular degradation of small, foreign RNAs and to develop more stable ribozymes, the ability of Escherichia coli RNase mutants to digest ribozymes was examined. In soluble extracts, most (80 to 90%) of the endonucleolytic activity was due to RNases I and I*, since degradative activity was inhibited by Mg2+ and by the rna-2 mutation. Degradation by exonucleolytic activities was temperature sensitive in extracts from an rna pnp rnb(Ts) triple mutant but not in extracts from an rna rnb(Ts) double mutant. Thus, the products of rnb and pnp, RNase II and polynucleotide phosphorylase, respectively, appear to be the major exonucleases that degrade hammerhead ribozymes. Examination of intracellular degradation revealed that RNases I and I* contributed to about half of the degradative activity as judged by comparison of the rate of ribozyme decay in wild-type and rna-2 mutant cells. Little additional effect was observed in rne(RNase E) and rnc (RNaseIII) mutants. Taken together, these data indicate that hammerhead ribozymes are digested largely by the degradative class of RNase (RNases I, I* and II and polynucleotide phosphorylase).
In order to study the relation of ethmoid labyrinth to the adjacent structures, continuous axial CT scan of the ethmoid labyrinth was performed on 9 normal subjects. Meanwhile, coronal and axial CT scans of ethmoid labyrinth were obtained in 20 normal subjects. On an average, 22 scan sections were done on each subject. The relation of ethmoid labyrinth to adjacent structures was observed and the remarkable intersubject variations in this anatomic area were noted. The relation of posterior ethmoid cells or sphenoid sinus to the optic canal, and the relation of internal carotid artery to the sphenoid sinus were clearly demonstrated on axial scans. The detailed anatomic structure of the ostiomeatal complex and the anatomic relation of ethmoid labyrinth to orbit and anterior skull base were revealed by coronal CT scans. CT scan of the paranasal sinuses was of great importance in clinical practice for the purpose of etiological analysis of sinusitis, design of endoscopic sinus surgery and prevention of complication.
The Sac7 proteins from the hyperthermophile Sulfolobus acidocaldarius are a heterogeneous mixture of small, thermostable, nonspecific DNA-binding proteins. One of these proteins, Sac7d, has been overexpressed in Escherichia coli to provide a homogeneous preparation for structure, stability, and function studies. We present here essentially complete sequence-specific 1H NMR assignments for Sac7d, a delineation of secondary structural elements, and the high-resolution solution structure obtained from a full relaxation matrix refinement. The final structure provides an excellent fit to the NMR data with an NOE R-factor of 0.27 for backbone NOEs. The structure has a compact globular fold with 82% of the sequence involved in regular secondary structure: an antiparallel two-stranded beta-ribbon with a tight turn, followed by a short 3(10) helix, an antiparallel three-stranded beta-sheet, another short 3(10) helix, and finally four turns of alpha-helix. The amphipathic alpha-helix packs across the hydrophobic face of the three-stranded beta-sheet in an open-faced sandwich arrangement with at least one turn of the helix exposed beyond the sheet. The hydrophobic face of the beta-ribbon packs against a corner of the twisted beta-sheet. The single tryptophan responsible for the 88% fluorescence quenching upon DNA binding is exposed on the surface of the three-stranded beta-sheet. Lysines 5 and 7, whose monomethylation may be associated with enhanced thermostability, are highly solvent exposed along the inner edge of the two-stranded ribbon. The structure of Sac7d differs in many respects from that reported for the homologous native Sso7d [Baumann et al. (1994) Nature Struct. Biol. 1, 808] with a backbone RMSD greater than 3.0 A, largely due to the packing and length of the C-terminal alpha-helix which may be important in Sac7d DNA binding.
Sodium salicylate has the unusual property of partially inducing the human heat shock response (Jurivich, D. A., Sistonen, L., Kroes, R., and Morimoto, R. I. (1992) Science 255, 1243-1245). Salicylate induces the DNA binding state of the human heat shock transcription factor (HSF), but this is insufficient to elevate heat shock gene expression. Because it is not known how HSF enhances heat shock gene expression, further analysis of the transcriptionally inert, salicylate-induced HSF was undertaken to potentially identify components of the heat shock response that are necessary for full transcriptional induction. Like thermal stress, exposure of HeLa cells to salicylate led to the induction of HSF1 into a DNA-bound state. Despite continued exposure of cells to salicylate, HSF1.DNA binding attenuated much more rapidly than a continuous heat shock. Western blot analysis revealed that the salicylate-induced form of HSF1 was not hyperphosphorylated like the heat-induced form. Furthermore, supershifts of the HSF1 bound to an heat shock element (HSE) oligonucleotide by monoclonal antibodies to phosphoamino acids revealed that salicylate induced threonine phosphorylation of HSF1, whereas heat led to a predominance of HSF1 serine phosphorylation. These data suggest that salicylate-independent signals are necessary to convert HSF1 into a transactivator of heat shock gene expression and that brief acquisition of DNA binding by this factor is insufficient to maximally enhance transcription.
The CDKN2 gene, encoding the cyclin dependent kinase inhibitor p16, is a tumour suppressor gene involved in melanoma and maps to chromosome band 9p22. Mutations or interstitial deletions of this gene have been found both in the germline of familial melanoma cases and somatically in melanoma cell lines. Previous mutation analyses of melanoma cell lines have indicated a high frequency of C:G to T:A transitions, with all of these mutations occurring at dipyrimidine sites. Including three melanoma cell lines carrying tandem CC to TT mutations, the spectrum of mutations so far reported indicates a possible role for u.v. radiation in the mutagenesis of this gene in some tumours. To further examine this hypothesis we have characterised mutations of the CDKN2 gene in 30 melanoma cell lines. Nineteen lines carried complete or partial homozygous deletions of the gene. Of the remaining cell lines, eight were shown by direct sequencing of PCR products from exon 1 and exon 2 to carry a total of nine different mutations of CDKN2. Two cell lines carried tandem CC to TT mutations and a high rate of C:G to T:A transitions was observed. This study provides further evidence for the role of u.v. light in the genesis of melanoma, with one target being the CDKN2 tumour suppressor gene.
One hundred anterior branches of the lateral cutaneous nerve of the thigh were dissected and measured in 50 adult cadavers. The results show that this nerve was present in all sides, and the average length from the point of the nerve piercing the fascia lata to the point of its first branch was about 79 mm, the average width was 2.25 mm, and the average thickness was 0.79 mm. It was found that the nerve was approximately along the line between the anterior superior iliac spine and the midpoint of the upper margin of the patella. This study presents a simple method of locating this nerve for clinical reference. The results also reveal that this nerve is a possible donor for facial nerve repair.
Cellular oncogenes have been implicated in head and neck cancer development since 1986. More recently interest has focused on chromosome 11q13; oncogenes therein undergoing ongoing investigation include Bcl-1/Prad-1, Hst-1 and Int-2. Our laboratory has studied the Int-2 oncogene for several years, primarily in the breast. This paper presents our investigations of Int-2 in the head and neck. Thirty-four paraffin-embedded primary squamous cell carcinomas were studied for Int-2 gene amplification using a carefully controlled method of sequence quantification by DNA dot blots. Amplification, mostly low level, was identified in 62 per cent of samples studied. No clinical correlation to amplification could be found. Further studies are underway looking for evidence of expression of Int-2 in fresh tissues and for amplification and expression of other oncogenes on this amplicon.