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Biomedical subjects

Q Shi

Publications and source records attributed to Q Shi.

At least 163 records · Page 9Linked to original sources
PubMed1998

Polymorphism of DXS102 locus in Chinese population and its application to gene diagnosis in hemophilia B family.

OBJECTIVE: To establish the polymorphism of DXS102 locus from Xq26.3-27.1 in Chinese population for the gene diagnosis in Hemophilia B family. METHODS: DNA was extracted from blood samples obtained from Shanghai unrelated volunteer donors with phenol-chloroform method. A total of 23x chromosomes (154 from females, 80 from males) were studied. A hemophilia B family in which a hemophilia B patient has received gene therapy was analyzed. The polymorphism of DXS102 locus in Chinese population was determined with amplified fragment length polymorphisms assay (Amp-FLP), denaturing polyacrylamide gel electrophoresis, silver stain detection. Short tandem repeats (STRs) linkage analysis was used to conduct gene diagnosis in hemophilia B family. RESULTS: Eight alleles were found at DXS102 locus, of which two alleles were first reported. The repeated number of AC dinucleotide ranges from 13 to 21. And the values of the observed heterozygosity, calculated heterozygosity and polymorphism information content(PIC) were 0.87, 0.80, 0.80 respectively. It was also found that the difference of the allele frequencies of DXS102 in Chinese and European populations was significant. By using the linkage analysis of the DXS102 locus, a family with a hemophilia B patient receiving gene therapy in 1994 was analyzed and meanwhile a carrier in that family was then detected. CONCLUSIONS: The polymorphism of DXS102 locus reveals significant difference between Chinese and European populations. DXS102 locus can be used as a promising marker for gene diagnosis in hemophilia B family.

Alleles↗
PubMed1998

[DNA content, expression of proliferating cell nuclear antigen and surgical margins in locally advanced laryngeal cancer].

OBJECTIVE: To probe into the relationship between DNA content, PCNA and prognosis. METHODS: Surgical margins in 34 patients with locally advanced laryngeal cancers were observed by means of antibody to PCNA and DNA content measurements. RESULTS: The results demonstrated that at neoplasm margins of 1.0, 0.5, and 0 cm, PCNA indices were 8.62%, 17.76% and 50.32% and DNA indices were 0.98, 1.082 and 1.436 respectively. There is a significant difference between 1.0, 0.5 and 0 cm. CONCLUSION: Moderate and severe atypical hyperplasia of epithelium (AHE) was different from mild AHE and simple hyperplasia of epithelium in the biologic behavior of fission, proliferation and so on and surgical margins should be kept outside the area of moderate AHE.

Adult↗
PubMed1998

[Investigation on resources and commodities of Chinese medicinal herbs of Jili].

OBJECTIVE: To make clear the current situation of application of the Chinese medicinal herbs of Jili. METHOD: The medicinal resources and commodities of the berbs in some provinces and cities in China were investigated. RESULT: The species Tribulus terrestris included in is widely distributed in China. The fruit of T. terrestris which is widely used in China as a traditional medicine is commonly called Yingjili. Being only the fruit of Atriplex entralasiatica Ruanjili is produced mainly in Shandong Province and customarily used as Fructus Tribuli in some areas of Shandong and Hebei Provinces. CONCLUSION: The current situation of medicinal application of Jili basically conforms to the requirements of . The main variety is Yingjili.

China↗
PubMed1998

[Study on the supercritical-CO2 fluid extraction of Prunus mandshurica oils].

The paper reported technological study on extracting Prunus mandshurica oils by supercritical-CO2 fluid, mainly researched the influence of pressure, temperature, time and flow rate of CO2 on the oil yield, determined optimum technology of extracting the oils, compared the oils from SFE-CO2 and traditional technology.

Carbon Dioxide↗
PubMed1998

[Parental origin of extra chromosome 21 in Down syndrome detected by using short tandem repeat DNA polymorphisms after PCR amplification].

OBJECTIVE: To detect the parental origin of extra chromosome 21 in Down syndrome, using two (GT)n polymorphic markers-D21S215 and D21S120. METHODS: The alleles of D21S215 and D21S120 were amplified by polymerase chain reaction and identified with denaturing polyacrylamide gel electrophoresis followed with Ag-staining. The parental origin of extra chromosome 21 was determined by comparing genotypes of probands with their parents. RESULTS: The parental origin of extra chromosome 21 was determined in 17 Down syndrome out of 24, with 12 and 5 inherited from mother and father respectively. CONCLUSION: The parental origin of extra chromosome 21 in Down syndrome can be determined by analyzing the polymorphisms of D21S215 and D21S120. Etiological study of trisomy 21 should be focused on maternal meiosis.

Chromosomes, Human, Pair 21↗
PubMed1998

Interaction of ethanol and nitric oxide in the hypothalamic-pituitary-gonadal axis in the male rat.

Ethanol (EtOH) exerts deleterious actions on reproductive function at all three levels: the hypothalamus, pituitary, and gonad (HPG). Nitric oxide (NO), a newly identified messenger molecular in a variety of biological systems, has been suggested as playing a role in HPG hormone regulation. NO stimulates luteinizing hormone releasing hormone secretion from the hypothalamus and has variable effects on luteinizing hormone release from the pituitary. NO is inhibitory to testosterone production, and it may also directly inhibit some steroidogenic enzymes. Related studies in the accompanying paper have demonstrated that inhibiting NO synthase (NOS) using various NOS inhibitors can prevent the EtOH-induced suppression of testosterone on the male HPG axis, and this action is mainly, although not entirely, due to a direct gonadal effect. To further investigate the role of NO in the HPG axis, we assessed the HPG NO-NOS system by determining NOS mRNA levels, protein levels, and enzyme activity in the presence and absence of EtOH. At the testicular level, EtOH's action did not appear to be mediated by increasing NO content. However, EtOH was able to potentiate NO's suppressive effect on the testicular synthesis system. One locus where EtOH and NO interacted was at the steroidogenic enzyme level. N(G)-nitro-L-arginine methyl ester, a NOS inhibitor, was found to antagonize the EtOH-induced fall on P-450 17alpha-hydroxylase/C17-20 lyase mRNA levels when administered along with EtOH. EtOH had no apparent effect on the pituitary NO-NOS system and its effects on the hypothalamic NO-NOS system do not explain its ability to reduce luteinizing hormone releasing hormone secretion.

Animals↗
PubMed1998

Effect of nitric oxide synthase inhibitors on preventing ethanol-induced suppression of the hypothalamic-pituitary-gonadal axis in the male rat.

Ethanol (EtOH) suppression of the hypothalamic-pituitary-gonadal (HPG) axis results in broad reproductive malfunction. In the HPG axis, the suppressive effects of EtOH are manifested by decreased serum testosterone, reduced testicular luteinizing hormone (LH) receptor numbers, lowered serum LH and pituitary beta-LH mRNA levels (in castrated animals), and impaired luteinizing hormone releasing hormone (LHRH) release from the hypothalamus. Increasing evidence has suggested that nitric oxide (NO) plays a role in regulation of the HPG axis. NO was shown to stimulate LHRH secretion from the hypothalamus and to have variable effects on LH release from the pituitary. At the gonadal level, NO is inhibitory to testosterone production. NO may directly inhibit some testicular steroidogenic enzymes. To investigate the effect of EtOH, NO, and their interaction on the male HPG axis, three NO synthase (NOS) inhibitors, N(G)-nitro-L-arginine methyl ester, N(G)-nitro-L-arginine, and 7-nitro indazole were used to study overall HPG function in the presence and absence of EtOH. Animals were given intraperitoneal injections of saline, EtOH, various NOS inhibitors, or EtOH, along with NOS inhibitors 2 hr before sacrifice. Serum testosterone and LH concentrations, pituitary beta-LH mRNA levels, hypothalamic LHRH mRNA levels, and LHRH content were determined. It was found that blocking NOS by these NOS inhibitors prevented EtOH-induced suppression of testosterone and, in some cases, serum LH. However, this was not accompanied by concurrent changes with NOS blockade on LHRH mRNA, hypothalamic pro-LHRH or LHRH content or pituitary LH beta mRNA levels. It appears that the protective effect of NOS blockade was largely, although not completely, due to a direct effect at the gonadal level.

Animals↗
PubMed1997

Antitumor agents. 172. Synthesis and biological evaluation of novel deacetamidothiocolchicin-7-ols and ester analogs as antitubulin agents.

A series of novel 7-O-substituted deacetamidothiocolchicine derivatives has been synthesized and evaluated for their inhibitory activity against tubulin polymerization, the binding of [3H]-colchicine to tubulin, and the growth of human Burkitt lymphoma cells. Of these new derivatives, thiocolchicone (8), wherein an acetamido group in thiocolchicine is replaced by a carbonyl oxygen at C(7), was obtained from deacetythiocolchicine (6) by Schiffs base equilibration and acid hydrolysis. Reduction of thiocolchiocone with sodium borohydride yielded the racemic alcohol 9, the structure of which was verified by X-ray crystallographic analysis. Optically pure alcohols 9a,b were obtained by treatment of 9 with the optically pure reagent (1S)-(-)-camphanic chloride followed by chromatographic separation of the camphanate esters and hydrolysis of the diastereomers. X-ray crystallographic analysis established the aS,7S-configuration of 9a. Racemic and optically active esters 11-15, 11a,b, 12a, 14a, and 15a were obtained by esterification of the corresponding alcohols. The compounds showing activity equivalent to or greater than (-)-thiocolchicione (2a) in all the biological assays were three (-)-aS,7S optically pure enantiomers: the alcohol 9a, the acetate 11a (an oxygen isostere of thiocolchicine), and the isonicotinoate 15a. In addition, the ketone 8 and two (-)-aS,7S enantiomers (12a, 14a) had high activity in the biochemical assays with tubulin but reduced antiproliferative activity. In all cases, optically pure isomers with the (-)-aS,7S configuration exhibited greater biological activity than racemic mixtures or isomers or isomers with the (+)-aR,7R configuration.

Antineoplastic Agents↗
PubMed1997

Modulation of the specificity and activity of a cellular promoter in an adenoviral vector.

Most gene therapy studies with recombinant adenoviruses employ viral promoters and lack tissue specificity. To determine whether a tissue-specific cellular promoter inserted into the adenoviral genome can direct the expression of a reporter gene in a tissue-specific manner, recombinant adenoviruses containing a nuclear lacZ gene driven by a human ventricular/slow muscle myosin light chain 1 promoter with and without a muscle creatine kinase enhancer were constructed. The ability of these viruses to express the reporter genes in infected myogenic and nonmyogenic cell lines was studied. Intramuscular injection of these viruses into mice showed that little or no reporter gene expression occurred in muscle fibers, although a relatively high level of lacZ gene expression was observed in surrounding connective tissue. Insertion of adenovirus sequences from the 5' inverted terminal repeat (ITR) region and/or the protein IX region into plasmids resulted in decreased reporter gene expression from myosin light chain 1 promoter in transfected C2C12 myotubes and 293 cells, as well as in injected muscles. These results suggested that negative elements are present in the adenoviral genome. This negative effect seems neither tissue nor species specific. Adenovirus cis-elements that may affect the specificity and activity of a cellular promoter are discussed.

Adenoviridae↗
PubMed1997

Accelerated endothelialization model for the study of Dacron graft healing.

Accelerated endothelialization was studied by creating a vascular tissue environment around porous Dacron grafts. Three study groups, each containing 10 dogs, were divided equally into 2- and 4-week implant periods, with 8 mm x 6 cm knitted Dacron grafts implanted in the abdominal aorta. Grafts in group 1, the control group, were implanted conventionally. In group 2 each implanted graft was completely wrapped in a resected segment of the autogenous inferior vena cava, with its intima against the wall. In group 3 the adventitial side of the vein was wrapped against the wall. The vein wrap produced accelerated endothelialization as follows: endothelial-like cell coverage scores at 2 and 4 weeks were, respectively, 78% and 98% for group 2 and 80% and 95% for group 3, compared to only 14% and 50% for group 1 (p < 0.05). The neointima, which contained smooth muscle cells, was formed as early as 2 weeks in the vein-wrapped grafts. There were no differences in the speed of healing or in healing patterns according to whether the intimal or the adventitial side of the inferior vena cava was placed against the graft. Histologic findings did not support the hypothesis that accelerated flow surface endothelialization results in direct migration of endothelial cells from the intima of the vein wrap, and there was no clear correlation between the surface endothelial-like cell coverage and microostia. To gain further insight into why accelerated healing occurs in this model, earlier observations accompanied by molecular biology analysis are needed, and vein wrap studies provide a method of comparison for this work.

Animals↗
PubMed1997

Early presence of endothelial-like cells on the flow surface of porous arterial prostheses implanted in the descending thoracic aorta of the dog.

The purpose of this study was to investigate early arterial graft healing and its sources in porous Dacron prostheses after very short implantation periods in the dog, using extensive histologic examination of serial sections. Preclotted Dacron prostheses 6 cm long and 8 mm in diameter were implanted in the descending thoracic aorta (DTA) of 14 dogs, and retrieved at 7 days (n = 6), 10 days (n = 4), and 14 days (n = 4). The flow surface was assessed for thrombus coverage, endothelial-like cell (ELC) coverage, and the number of microvessel ostia. Where an ELC island was identified under the stereomicroscope, full-wall longitudinal tissue samples were taken, and embedded in resin for light microscopy study of 6-microns, H&E-stained serial sections to determine general healing and interstitial tissue presence. If there was more than one ELC island, another full-wall sample was taken and embedded in paraffin for staining with laminin, collagen IV, and smooth muscle alpha-actin antibodies, and PTAH. All grafts were patent with very little thrombus. Islands of endothelial-like cells were found for each time period, and on all 14-day grafts. Endothelial-like cell coverage was highest at 14 days. On 7- and 10-day grafts, cells proved to be endothelium were found in the middle of the flow surface, unconnected to either pannus or perigraft tissue ingrowth. Healing occurs as early as 7 days in porous knitted Dacron grafts. The source at periods earlier than 10 days appears to be cells from the blood stream.

Animals↗
PubMed1997

Removal of endotoxin from recombinant protein preparations.

OBJECTIVES: To develop an effective method to remove endotoxin from large scale E. coli recombinant protein purifications. DESIGN AND METHODS: Triton X-114 phase separation, affinity chromatography utilizing immobilized polymyxin B or immobilized histidine, were used to remove endotoxin from purified preparations of recombinant CK-BB, CK-MB, CK-MM, myoglobin, and cardiac troponin I. Endotoxin levels were measured by a Limulus Amebocyte Lysate gel-clot assay. The immunoactivity of these protein preparations was determined by BIAcore analysis using a panel of in-house generated monoclonal antibodies and by a Stratus Fluorometric Analyzer. In the case of troponin I, the BIAcore was also utilized to measure troponin C interactions. RESULTS: Phase separation with Triton X-114 was the most effective method in reducing the amount of endotoxin present in the protein preparations compared to either polymyxin B or histidine affinity chromatography. With Triton X-114, the reduction in endotoxin levels was greater than 99% and recovery of the proteins after endotoxin removal was greater than 90%. All three procedures for removing endotoxin had no deleterious effects on the immunoactivity of majority proteins when tested with a panel of monoclonal antibodies. Troponin I also retained its ability to bind to troponin C in the presence of Ca2+. Recombinant CK-BB and CK-MM which were expressed in the soluble fraction of E. coli cell lysates, contained significantly higher endotoxin levels than recombinant CK-MB, myoglobin and cardiac troponin I which were expressed in the form of inclusion bodies. CONCLUSION: Of the three methods tested, Triton X-114 phase separation was the most effective way of removing endotoxin from recombinant proteins.

Biochemistry↗
PubMed1997

Analytical performance and clinical utility of a sensitive immunoassay for determination of human cardiac troponin I.

OBJECTIVES: To determine the serum and plasma level of human cardiac troponin I (cTnI) resulting from myocardial damage, we have developed a sensitive and specific one-step enzyme immunoassay to measure cardiac troponin I. DESIGN AND METHODS: The COBAS cTnI assay is a semi-automated one-step solid phase immunoassay compatible with the COBAS Core. The assay is performed in a sandwich type format using a polyclonal goat antibody capture and two highly specific horseradish peroxidase conjugated monoclonal antibody detectors directed against different epitopes of the cTnI molecule. Calibrators were made with purified recombinant cTnI. RESULTS: The level of cTnI was determined in 84 healthy donors with no evidence of myocardial injury, resulting in a lower limit of detection (LLD) of 0.09 microgram/L. The upper reference limit (URL) of the normal reference range was calculated as 0.20 microgram/L. The dynamic range of the consequent EIA was between 0.09 and 6.0 micrograms/L with a total assay time of 45 min. Intra-assay and inter-assay variances (CVs) were < or = 4%. Cross-reactivity with fast and slow skeletal troponin I was absent in concentrations up to 2.0 mg/L. Common interferents yielded negative results in the cTnI assay. Clinical utility was confirmed by measuring the circulating serum or plasma levels of cardiac troponin I in serial samples from marathon runners, clinical samples from trauma patients, and patients presenting to the Emergency Department with complaints of chest pain. Results were further evaluated using clinical diagnosis at discharge and quantified concentrations of other cardiac markers by a Stratus analyzer and ELISA procedures. CONCLUSIONS: Results from normal and clinical samples assayed in house for cTnI concentrations indicate that the Spectral EIA is a highly sensitive means of quantifying cTnI levels in serum and plasma for acute cardiac syndrome. The cardiac specificity of cTnI over other well-known cardiac markers is reflected in experimental results and parallel clinical diagnosis.

Animals↗
PubMed1997

Effect of altered blood flow on the caliber and morphology of the internal thoracic artery in the dog.

OBJECTIVE: The purpose of this study was to evaluate in dogs the effect of blood flow alteration on caliber and morphology of in situ internal thoracic arteries. METHODS: Six dogs underwent creation of a unilateral distal arteriovenous fistula between the internal thoracic artery and vein at the sixth rib to create high flow, and in six others the internal thoracic artery was unilaterally skeletonized and dissected. For both groups the contralateral internal thoracic artery served as the control; sides were alternated among cases. Blood flow was measured for shear stress calculation before and after surgical alteration. After 2 months, internal thoracic arteries were harvested with the entire anterior chest plate, which was dynamically inflated and fixed with 10% formalin at a controlled pressure of 120 mm Hg after angiography had been done at the same pressure. The luminal diameters were then measured at eight levels on the angiograms. Arterial tissue samples were taken at three levels and embedded, sectioned, and treated with hematoxylin-eosin and Verhoeff-van Gieson stains. Digital imaging analysis was used for quantitative morphometric studies. RESULTS: All fistulas remained patent. In comparison with control arteries, high-flow internal thoracic arteries dilated and low-flow internal thoracic arteries narrowed, which was associated with significant change in shear stress for both groups. There were no substantial structural changes in the walls of either group. CONCLUSION: In the dog, the luminal diameter of the internal thoracic artery responds to altered blood flow without intimal thickening or other undesirable wall changes.

Animals↗
PubMed1997

Implant site influence on arterial prosthesis healing: a comparative study with a triple implantation model in the same dog.

PURPOSE: The purpose of this study was to develop a cost-effective canine graft healing model that gives information on various implant sites and controls for variable factors between graft locations and between animals and to compare the influence of implant site (retropleural, retroperitoneal, and subcutaneous areas) on arterial graft healing in the same subject under such controlled study conditions. METHODS: Five mongrel dogs were studied for 8 weeks, and one was studied for 3 years. Each received three porous Dacron grafts during the same surgery: a carotid-femoral bypass (C-FB) and interposition grafts in the descending thoracic aorta and abdominal aorta. To produce comparable shear stress calibers of the C-FB and abdominal aorta grafts were 2 mm less than those of the descending thoracic aorta, and a distal arterio-venous fistula was created to further increase the C-FB flow. For comparable blood aggregation status platelet aggregation was preevaluated and adjusted with antiplatelet agents. Graft flow surfaces were assessed for thrombus-free surface and endothelial-like cell coverage scores. Tissue samples were studied with hematoxylin-eosin, factor VIII/ von Willebrand factor, smooth muscle alpha-actin staining, and scanning electron microscopy and transmission electron microscopy. RESULTS: All grafts were patent. Shear stress for the three grafts and platelet aggregation among the study subjects were comparable. Healing of descending thoracic aorta and abdominal aorta grafts was similar, but C-FB healing was slow, incomplete, and uneven, with a high incidence of seroma. Eight-week and 3-year results were comparable. CONCLUSIONS: This model gives broad healing information about the areas where grafts are often implanted in humans. Eight weeks appears to be a sufficient period to reflect basic and general healing characteristics. Grafts heal better in the retropleural and retroperitoneal areas than in the subcutaneous tissues.

Animals↗
PubMed1997

Endothelium on the flow surface of human aortic Dacron vascular grafts.

PURPOSE: We have previously observed endothelium on two human vascular prostheses explanted under optimal conditions for flow surface preservation. In this study we sought to further verify the hypothesis that endothelialization can occur on clinical grafts and that it can be detected in specimens that have been promptly removed and properly preserved in a timely manner. METHODS: We studied 29 aortic grafts. Of these, 11 Dacron bypass grafts were in a condition suitable for analysis with light microscopy and immunocytochemistry staining, and scanning and transmission electron microscopy. RESULTS: Three grafts had endothelium beyond the pannus, identified by factor VIII/ vWF, Ulex europaeus agglutinin, and collagen IV positivity. Specimen A, a knitted 6-year implant, was preserved by embalming 3 hours after the patient's death and had a firmly attached outer capsule with fibroblasts, collagen, giant cells, and microvessels in the interstices. Specimen B, a woven 18-year implant, was retrieved at reoperation and immediately fixed in 10% formalin; it had no outer capsule and no tissue ingrowth. Specimen C, a woven 7-year implant, was removed and fixed 5 hours after the patient's death; it had a firmly attached outer capsule but no tissue ingrowth beyond the outer portion of the wall. CONCLUSIONS: The rapidity with which the specimens were fixed probably enabled identification of endothelium. These findings suggest that endothelialization of synthetic arterial grafts may occur more frequently in human beings than previously recognized.

Aged↗
PubMed1997

Antitumor agents--CLXXV. Anti-tubulin action of (+)-thiocolchicine prepared by partial synthesis.

(+)-Thiocolchicine (2b) was prepared from (+/-)-colchicine (1) in a five-step reaction sequence that included chromatographic separation of appropriate camphanylated diastereomers. Acid hydrolysis of the (+)-diastereomer, followed by acetylation, yielded the desired product 2b. (+)-Thiocolchicine has 15-fold lower inhibitory activity against tubulin polymerization than (-)-thiocolchicine, and is 29-fold less potent for inhibiting growth of human Burkitt lymphoma cells. The enantiomer 2a, prepared from the (-)-camphanylated diastereomer, had potent activity in all assays comparable to that of (-)-thiocolchicine prepared by other methods. These results support the hypothesis that the proper configuration of colchicine-related compounds is an important requirement for their anti-tubulin action.

Antineoplastic Agents↗