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Biomedical subjects

M S Li

Publications and source records attributed to M S Li.

At least 37 records · Page 2Linked to original sources
PubMed1996

The Vpr protein of human immunodeficiency virus type 1 binds to nucleocapsid protein p7 in vitro.

The Vpr protein of human immunodeficiency virus type 1 (HIV-1) is incorporated into the virion by the Gag polyprotein precursor Pr55gag. The importance of the p6gag sequence at the C-terminal end of Pr55gag has a crucial role in Vpr incorporation. To identify the Gag sequences directly involved in Vpr binding, we compared the Vpr binding affinities of the 71 amino acid nucleocapsid protein p7, the C-terminal peptide (35-71) p7C and p6gag by affinity chromatography. p7 and p7C have the strongest Vpr binding activities compared to p6gag. These results suggest that the nucleocapsid protein and its C-terminal domain may be important for the incorporation of Vpr into the mature HIV-1 virion and the subsequent localisation of viral nucleic acid to the cell nucleus by Vpr.

Blotting, Western↗
PubMed1996

Topical glucocorticoids modulate the lesion interface after cerebral cortical stab wounds in adult rats.

A lesion interface, consisting of a glia limitans lined by a laminin-rich basal lamina and leptomeningeal cells, forms within 2-3 weeks after penetrating wounds to the adult mammalian central nervous system (CNS). This interface prevents the growth of axons across the lesion. We have examined the effects of topically applied steroids on the formation of such an interface after stab wounds to the adult rat cerebral cortex. Immediately after lesioning, the surface of cortex in the region of the wound was treated with a topical application of either 0.1% halcinonide or 0.05% betamethasone dipropionate or their respective placebos. Cryostat sections through the lesioned area were obtained 3 weeks later and assessed by immunofluorescence. Steroid treatment attenuated all components of the lesion. The continuous anti-laminin labeling along the lesion in untreated rats became patchy after steroid treatment. The number of leptomeningeal cells that infiltrated into the wound was reduced in the laminin-negative regions in steroid-treated rats. In addition, astrocytic processes in the laminin-negative regions after steroid treatment were loosely arranged, compared with the tightly packed parallel processes forming the glia limitans in laminin-positive regions in controls. The mechanism of steroid-mediated attenuation of the lesion interface was examined in vitro. Betamethasone but not halcinonide reduced laminin secretion slightly in leptomeningeal cell cultures, but both steroids reduced cell proliferation. These results suggest that steroids modulate the formation of the lesion interface after CNS injury, at least in part by decreasing leptomeningeal cell proliferation. Such modulation of the lesion interface by steroids or other agents may permit the growth of axons across the lesion site and thus could enhance the overall degree of axon regeneration if other factors such as neurotrophic support and neutralization of axon growth inhibitory molecules are optimized.

Animals↗
PubMed1995

Human eosinophil major basic protein, a mediator of allergic inflammation, is expressed by alternative splicing from two promoters.

Human eosinophil major basic protein (MBP) is one of the principal mediators of injury to parasites and tissues in allergic inflammation. MBP is stored in eosinophil crystalloid granules and released with other granule constituents during eosinophil action. Previous studies have identified an MBP gene promoter that generates a 1.0 kb mRNA transcript encoding MBP preproprotein which undergoes processing to the mature storage form. To investigate how the MBP gene is regulated, we have examined the identity and levels of the MBP transcripts both in precursor cells and in blood eosinophils. It was found that the gene was expressed from two upstream promoters, a distal promoter P1 in addition to the previously described promoter P2. Evidence for the second promoter was initially provided by isolation from a human HL-60 leukaemic cell cDNA library of a novel 1.6 kb MBP cDNA that was distinct from the known 1.0 kb cDNA. The complete nucleotide sequence of the 1.6 kb cDNA was determined, and showed that the two cDNAs had identical coding and 3' untranslated regions but differed in their 5' sequences. By isolating and sequencing MBP genomic clones from an arrayed chromosome 11 library, it was demonstrated that the MBP gene is composed of nine upstream exons and five coding exons. The 1.6 and 1.0 kb cDNAs arise by differential splicing of alternate MBP transcripts from promoters P1 and P2 respectively, located 32 kb apart in the genomic DNA. Primer extension analysis identified two transcription start sites at P1, neither associated with a typical TATA box motif. Northern blotting and reverse-transcription PCR analysis showed that the 1.0 kb mRNA was present at higher levels than the 1.6 kb species in immature cells including HL-60 and bone-marrow cells. By contrast, low levels of 1.6 kb mRNA transcripts predominated in differentiated blood eosinophils. The results are compatible with differential use of P1 and P2 promoters as a mechanism for regulation of MBP expression during eosinophil maturation.

Alternative Splicing↗
PubMed1995

Expression in Escherichia coli and purification of human eosinophil-derived neurotoxin with ribonuclease activity.

Eosinophil-derived neurotoxin (EDN) is a ribonuclease with neurotoxic and helminthotoxic properties. It is present in the crystalloid granules of human eosinophils. We report the expression and characterization of a functionally active recombinant human EDN using the pMAL-cRI expression system. A cDNA for mature EDN was obtained by PCR and inserted in pMAL-cRI downstream of the malE gene encoding maltose binding protein. Induction of the ptac promoter of the plasmid in Escherichia coli strain BL21(DE3) resulted in high level expression of soluble MAL-EDN fusion protein. Cleavage of affinity purified fusion protein with Factor Xa protease released recombinant EDN which comigrated with native EDN on SDS-polyacrylamide gels and cross-reacted with a polyclonal anti-EDN antiserum on Western blots. IN contrast to previous attempts at EDN expression, denatured and refolded EDN had ribonuclease activity and was prepared in microgram amounts. The availability of recombinant human EDN should facilitate studies of its structure and biological functions.

ATP-Binding Cassette Transporters↗
PubMed1994

In-vitro activity of quinolones and macrolides against mycobacteria.

The activities of eight quinolones (ciprofloxacin, clinafloxacin, levofloxacin, ofloxacin, A-80556, sparfloxacin, temafloxacin and tosufloxacin) and three macrolides (azithromycin, clarithromycin and erythromycin) against 98 clinical isolates of Mycobacterium tuberculosis and 120 isolates of five different atypical mycobacterial species including 20 Mycobacterium kansasii, 25 Mycobacterium scrofulaceum, 25 Mycobacterium avium/intracellulare, 25 Mycobacterium chelonae and 25 Mycobacterium fortuitum were determined with the Middlebrook 7H9 broth macrodilution method. Sparfloxacin, clinafloxacin, levofloxacin, ciprofloxacin and ofloxacin were active against M. tuberculosis (MIC90 0.06-0.5 mg/L; MBC90 0.125-2.0 mg/L). However, higher MIC90S and MBC90S of these quinolones were obtained for strains of multi-drug resistant M. tuberculosis. The macrolides tested had poor activity against M. tuberculosis isolates (MIC90 > 8.0 mg/L). Furthermore, high MIC90S of the quinolones and macrolides (2.0 to 8.0 mg/L) were obtained for clinical isolates of atypical mycobacteria, with the exception of clarithromycin against M. kansasii (MIC90 = 1.0 mg/L) and sparfloxacin against M. scrofulaceum (MIC90 = 1.0 mg/L).

4-Quinolones↗
PubMed1994

Nonpenetrating trauma to the carotid artery: seven cases and a literature review.

Nonpenetrating carotid trauma is uncommon and frequently missed on initial examination. The cases of seven patients seen over a period of 21 years are presented and 100 cases from the most recent literature are reviewed. Causes and mechanisms of injury, clinical presentation, investigations, management, and outcome are discussed. Causes of injury were three motor vehicle collisions, two falls, one sports injury, and one blow to the face. Clinical presentation was early in four and delayed in three. The earliest symptoms and signs were a change in mental status, headache, unprovoked fall, focal weakness, neglect, and dysphasia. Doppler studies may be useful in screening, but a definitive diagnosis is made with the help of angiography. Two patients were treated surgically; one died, one with delayed symptoms from a pseudoaneurysm recovered completely. Five patients were given anticoagulants; all survived with permanent deficits related to their pretreatment neurologic status. The outcome in 100 recent cases from the literature has improved compared with previous reports. The overall mortality was 12%. The outcome in our seven cases supports recent trends toward a strategy of early anticoagulation and selective surgical treatment.

Adult↗
PubMed1994

Interleukin-5 mRNA levels in blood and bone marrow mononuclear cells from patients with the idiopathic hypereosinophilic syndrome.

High blood eosinophil counts in humans are usually due to parasitic infections, allergic processes or malignant diseases. Interleukin-5 (IL-5) is thought to be the principal eosinopoietic stimulus in most of these patients. As the causes of persistent eosinophilia in patients with the idiopathic hypereosinophilic syndrome (HES) are (by definition) unknown, a semi-quantitative assay for IL-5 mRNA in eosinophils and mononuclear cells was carried out using samples from 11 patients with HES. In three patients, unstimulated peripheral blood mononuclear cells (PBMC) and bone marrow mononuclear cells (BMMC) had markedly raised levels of IL-5 mRNA. In four of five patients studied, phorbol myristate acetate (PMA) stimulation of PBMC induced them to produce more IL-5 mRNA than four healthy subjects. In five patients, bone marrow IL-5 mRNA levels were related to blood eosinophil counts. Blood eosinophils from all six patients tested, and bone marrow granulocytes from four patients had undetectable levels of IL-5 mRNA. It was concluded that HES can be separated into two groups. One has high levels of IL-5 mRNA and/or an enhanced IL-5 mRNA response to stimulation. They may well respond to treatments which inhibit the effects of this cytokine on eosinophil progenitor cells. The second group appears to have a disease which is IL-5 independent.

Adult↗
PubMed1994

Expression of lacto-N-fucopentaose III (CD15)- and sialyl-Lewis X-bearing molecules and their functional properties in eosinophils from patients with the idiopathic hypereosinophilic syndrome.

As the carbohydrate lacto-N-fucopentaose III (CD15 antigen or X-determinant) and its sialylated derivative sialyl-Lewis X are involved in the adhesion of cells rolling along the surface of endothelial cells, experiments were done to study the presence of these molecules on human eosinophils from patients with the idiopathic hypereosinophilic syndrome. Normal-density eosinophils from some patients showed higher levels of expression for lacto-N-fucopentaose III than light-density eosinophils. In contrast, sialyl-Lewis X was highly expressed by light-density eosinophils. Activation of normal-density eosinophils with calcium ionophore A23187 resulted in increased expression of these molecules for a short time. Monoclonal antibodies to these carbohydrates stimulated eosinophils to secrete eosinophil cationic protein, but not eosinophil peroxidase, and acted as costimulatory signals for C3b-induced degranulation of eosinophil cationic protein. It was suggested that CD15 and sialyl-Lewis X might contribute to eosinophil-mediated tissue injury in patients with the idiopathic hypereosinophilic syndrome.

Adult↗
PubMed1994

Evaluation of three culture media and their combinations for the isolation of Mycobacterium tuberculosis from pleural aspirates of patients with tuberculous pleurisy.

The abilities of three culture systems to detect Mycobacterium tuberculosis from 480 pleural aspirate specimens in 347 patients were compared. The systems were the Bactec 12B radiometric detection system (Bactec 12B), the Middlebrook 7H9 broth (7H9), and two solid media including one pyruvate enriched Lowenstein-Jensen slope (LJP) and one non-selective Middlebrook 7H11 agar slope (7H11). In the 116 specimens obtained from 85 patients of confirmed pleural tuberculosis for the isolation of Mycobacterium tuberculosis, the diagnostic yields were 33 (39%) cases from Bactec 12B, 24 (28%) cases from 7H9 and 21 (25%) cases from LJP/7H11, with median detection times of 18, 47.5 and 44 days respectively. Thirteen isolates were recovered by the Bactec 12B only, 9 by the 7H9 broth only, and 2 by the LJP/7H11 agar only. The use of a combination of the three culture systems substantially increased the diagnostic yield to 47% over the individual systems. These results show that a significantly greater and faster diagnostic yield can be obtained by including Bactec 12B in pleural aspirate cultures for the recovery of Mycobacterium tuberculosis.

Biopsy, Needle↗
PubMed1993

Screening of esophageal and gastric cancer by occult blood bead detector.

A previously described bead method used to detect blood is simple, cheap, and effective; it can be repeated periodically. In 11 provinces in China, 233,825 persons (age range, 30-70 years) were screened. A positive occult blood test result was found in 28,557 persons (12%). Of these, 16,918 underwent a gastroscopy, resulting in the detection of 581 cancers that were located in similar frequencies in the esophagus, gastric cardia, and gastric body; 70% of the lesions were in an early or moderately advanced stage. Among 119 patients with early-stage cancer, the 3-year survival rate was 98.3%. After preliminary screening of cancer, the 4-year follow-up found a mortality rate in the group with negative occult blood test results that was only 25% of that of the positive group. Preliminary yearly screening would detect more early cancers and fewer advanced cancers. The mortality rate of esophageal and gastric cancer might be reduced drastically.

Adult↗
PubMed1993

In vivo study of bleeding time and arterial hemorrhage in hypothermic versus normothermic animals.

This in vivo study confirmed impaired hemostasis during hypothermia in a swine model. Group I (normothermic, n = 8) and group II (hypothermic, n = 8) animals were anesthetized and instrumented for continuous peritoneal irrigation and monitoring of heart rate and blood pressure. The effects of hypothermia, hypotension, and inotrope on bleeding time and bleeding from two types of arterial injuries were evaluated. Our findings were that (1) bleeding time was significantly prolonged in hypothermic animals; (2) the differences in blood loss from partially torn artery (PTA) and completely cut artery (CCA) at both normothermic and hypothermic temperatures did not reach statistical significance; and (3) blood loss from PTA was greater than CCA when norepinephrine (Levophed) was infused to elevate blood pressure in hypotensive animals at normal core temperature.

Algorithms↗
PubMed1993

Postantibiotic effects of amikacin and ofloxacin on Mycobacterium fortuitum.

A study of postantibiotic effects (PAE) in vitro of amikacin and ofloxacin on Mycobacterium fortuitum isolates from sternotomy wounds by use of the dilution method for drug removal showed that both drugs exhibited good bactericidal activities, with the PAE of amikacin lasting from 13.5 to 27.6 h and the PAE of ofloxacin lasting from 1.2 to 5.0 h. These laboratory results concur with our experience of the efficacy of once-daily dosing with these drugs in the treatment of infections caused by M. fortuitum. These data may have therapeutic implications in guiding the scheduling of the administration of drugs in these infections, which require a long duration of therapy.

Amikacin↗
PubMed1993

Lack of activities of amikacin and sulphamethoxazole against Mycobacterium avium-intracellulare.

Twenty clinical isolates of Mycobacterium avium-intracellulare were tested with amikacin and sulphamethoxazole for in-vitro susceptibilities. The MICs and MBCs of the former drug ranged from 8 to > 64 mg/L (median MIC: 64 mg/L, median MBC: > 64 mg/L). The MICs and MBCs of the latter drug were found to be > 256 mg/L. Each of eight patients with invasive pulmonary disease due to these organisms was treated with amikacin for six months and with cotrimoxazole (sulphamethoxazole-trimethoprim) for one year. Only one patient had sustained bacteriological conversion. Three patients showed a transient reduction of bacillary load during the period of amikacin administration. The rest all failed to show response. Thus sulphamethoxazole was found to have no activity against Mycobacterium avium-intracellulare, and amikacin has doubtful activity when used alone in treatment of M. avium-intracellulare infection.

Adult↗
PubMed1991

[A clinical analysis of 317 cases of salpingo-anastomoses].

Influences of various factors on status pregnancy in 317 women having had salpingo-anastomoses were analyzed. All these cases, aged 21 to 45, had end to end operation under direct observation. Through 1-5 years' follow up, two of the 317 women were found to be restricted, 295 were pregnant or already delivered. The re-potent rate was 99.3%, while the re-pregnant rate 93.1%. No statistically obvious disparity in pregnancy rate was found as regard to age, tubal ligation technique site of anastomosis. However, there was influence over the pregnancy rate by the length of the remaining tube and oviduct inflammation. This paper also discussed on the anastomosis method, length of contraception period, observation period and the choice of stents.

Adult↗
PubMed1988

Screening for upper digestive tract cancer with an occult blood bead detector. Investigation of a normal north China population.

Screening of the general population for cancer of the upper digestive tract using an occult blood bead detector supplemented by fibergastroscopy was able to detect 126 cancers of the esophagus, gastric cardia, and stomach. This screening was carried out in two of the high-risk areas, Yangzhong and Wun counties, located in central eastern and north China. The occult blood bead detector, swallowed and retained in the stomach for 3 minutes, was able to sort out the suspected persons by its coloration from sky blue (+) to dark blue ( ) on removal. The subsequent fibergastroscopy could make a definite diagnosis by biopsy specimen. A preliminary trial performed on patients with established cancers was 100% (18 of 18) positive for stomach cancer and 81% (13 of 16) positive for esophageal cancer. A public survey of the high-risk areas, screening 38,073 normal persons ranging in age from 35 to 70 years, showed positive results in 9204 (24.2%) persons. Subsequent fibergastroscopy carried out in 4023 occult blood bead detector positive persons showed 126 cancers (41 of the esophagus, 46 of the gastric cardia, and 39 of the stomach). Factors influencing the cancer detection rate were sex, age, and degree of positivity of the occult blood bead detector test. Seventy per cent (89 of 126) of the cancers discovered were in an early stage. The occult blood bead detector costs the primary screened person only 30 cents and may be practical for the large-scale screening of cancer of the upper digestive tract in developing areas.

China↗