Search PubMed⌕ Search

Biomedical subjects

X Ma

Publications and source records attributed to X Ma.

At least 289 records · Page 16Linked to original sources
PubMed1996

Colocalization of cell division proteins FtsZ and FtsA to cytoskeletal structures in living Escherichia coli cells by using green fluorescent protein.

In the current model for bacterial cell division, FtsZ protein forms a ring that marks the division plane, creating a cytoskeletal framework for the subsequent action of other proteins such as FtsA. This putative protein complex ultimately generates the division septum. Herein we report that FtsZ and FtsA proteins tagged with green fluorescent protein (GEP) colocalize to division-site ring-like structures in living bacterial cells in a visible space between the segregated nucleoids. Cells with higher levels of FtsZ-GFP or with FtsA-GFP plus excess wild-type FtsZ were inhibited for cell division and often exhibited bright fluorescent spiral tubules that spanned the length of the filamentous cells. This suggests that FtsZ may switch from a septation-competent localized ring to an unlocalized spiral under some conditions and that FtsA can bind to FtsZ in both conformations. FtsZ-GFP also formed nonproductive but localized aggregates at a higher concentration that could represent FtsZ nucleation sites. The general domain structure of FtsZ-GFP resembles that of tubulin, since the C terminus of FtsZ is not required for polymerization but may regulate polymerization state. The N-terminal portion of Rhizobium FtsZ polymerized in Escherichia coli and appeared to copolymerize with E. coli FtsZ, suggesting a degree of interspecies functional conservation. Analysis of several deletions of FtsA-GFP suggests that multiple segments of FtsA are important for its localization to the FtsZ ring.

Adenosine Triphosphate↗
PubMed1996

Normalization of diabetes in spontaneously diabetic cynomologus monkeys by xenografts of microencapsulated porcine islets without immunosuppression.

Porcine pancreatic islets were microencapsulated in alginate-polylysine-alginate capsules and transplanted intraperitoneally into nine spontaneously diabetic monkeys. After one, two, or three transplants of 3-7 x 10(4) islets per recipient, seven of the monkeys became insulin independent for periods ranging from 120 to 804 d with fasting blood glucose levels in the normoglycemic range. Glucose clearance rates in the transplant recipients were significantly higher than before the graft administration and the insulin secretion during glucose tolerance tests was significantly higher compared with pretransplant tests. Porcine C-peptide was detected in all transplant recipients throughout their period of normoglycemia while none was found before the graft administration. Hemoglobin A1C levels dropped significantly within 2 mo after transplantation. While ketones were detected in the urine of all recipients before the graft administration, all experimental animals became ketone free 2 wk after transplantation. Capsules recovered from two recipients 3 mo after the restoration of normoglycemia were found physically intact with enclosed islets clearly visible. The capsules were free of cellular overgrowth. Examination of internal organs of two of the animals involved in our transplantation studies for the duration of 2 yr revealed no untoward effect of the extended presence of the microcapsules.

Animals↗
PubMed1996

Sequences outside the homeodomain of bicoid are required for protein-protein interaction.

The Drosophila morphogenetic protein Bicoid (Bcd) is required for the development of anterior structures of the embryo. Bcd, a homeodomain protein, is distributed as an anterior-to-posterior gradient in the embryo. It stimulates the expression of the hunchback (hb) gene in the anterior half in an all-or-none fashion. We have recently shown that Bcd binds cooperatively to a hb enhancer element and proposed that cooperative DNA binding is facilitated by an interaction between Bcd molecules. In this report, we further analyze the interaction between Bcd molecules and define regions important for protein-protein interaction. We show that the homeodomain of Bcd alone fails to interact with another Bcd molecule efficiently. The protein sequence flanking either side of the homeodomain restores the protein-protein interaction function. Mutations in the homeodomain that affect DNA binding do not adversely affect the protein-protein interaction function, suggesting that the surfaces for DNA binding and protein-protein interaction are separable. Finally, we demonstrate that the homeodomain of Bcd alone, unlike the intact Bcd, fails to bind DNA cooperatively. These results further support the notion that cooperative DNA binding is facilitated by the interaction between Bcd molecules. They strongly suggest that protein-protein interaction is an important property of Bcd for its biological activities.

DNA Footprinting↗
PubMed1996

Differential production of interleukin 10 during human immunodeficiency virus infection.

Interleukin 10, a product of T and B cells and monocytes, displays many Th2-like properties through inhibition of Th1 cell functions. Interleukin 10 is thought to play a major role in the immune dysfunction seen in HIV-infected individuals. In this study, we evaluated in detail the production of IL-10 during HIV infection. Although the constitutive production of IL-10 did not differ in PBMCs from healthy donors and HIV-infected individuals, IL-10 was differentially produced in response to polyclonal activators. The overall plasma IL-10 levels were similar in 32 controls and 67 patients at different stages of the disease and receiving different antiretroviral drugs. However, patients with low CD4 T cell count (< 200/mm3) secreted approximately three-fold more IL-10 than did patients with high CD4 T cell count (> 500/mm3). Competitive/quantitative PCR revealed similar levels of mRNA expression in PBMCs from controls and HIV-infected individuals. In vitro HIV infection rapidly and transiently induced IL-10 production in PBMCs and monocytes, and the low level of endogenously secreted IL-10 failed to inhibit HIV replication in acutely infected monocytes. On the other hand, HIV infection of selected CD4+ T cell clones generated in a Th1- or Th2-like environment, differentially up-regulated IL-10 production, with significantly higher production by Th2 clones. Together, our data indicate that IL-10 production is more complex than previously thought, and may depend on several factors such as producer cells, nature of the stimuli, as well as viral isolates.

CD4 Lymphocyte Count↗
PubMed1996

The retinoblastoma gene product inhibits TGF-beta1 induced apoptosis in primary rat hepatocytes and human HuH-7 hepatoma cells.

Transforming growth factor-beta1 (TGF-beta1) can induce rapid growth arrest and apoptosis in hepatic cells. Its growth suppressive effects appear to be linked to decreased phosphorylation of the protein product of the retinoblastoma gene, pRb. To characterize the role of pRb in apoptosis, we examined endogenous retinoblastoma gene (Rb) expression following treatment with TGF-beta1, okadaic acid, or antisense Rb S-oligonucleotides in cultured primary rat hepatocytes and human hepatoma HuH-7 cells. We also investigated the effects on apoptosis of Rb overexpression following transfection with vectors containing wild-type Rb in HuH-7 cells. Our results indicated that transfection with Rb antisense S-oligonucleotides blocked the expression of pRb in cultured primary hepatocytes and induced apoptosis. Treatment of HuH-7 cells with TGF-beta1 inhibited expression and phosphorylation of pRb, and also induced apoptosis. Furthermore, 93% of viable preapoptotic cells were arrested in the G1 phase of the cell cycle. Incubation with the phosphatase inhibitor okadaic acid maintained pRb in its phosphorylated state, and resulted in significant apoptosis. Overexpression of wild-type Rb inhibited TGF-beta1 induced apoptosis in HuH-7 cells. In contrast, overexpression of transcription factor E2F-1, a known target for the activity of pRb, caused significant apoptosis. However, coexpression of Rb suppressed E2F-1 induced apoptosis in HuH-7 cells. Our results suggest that inhibition of pRb expression is associated with hepatocyte apoptosis. Furthermore, E2F-1 appears to be a target in the pathway through which pRb modulates the apoptotic threshold in hepatic cells. Finally, the data suggest that these cells exit the cell cycle during the G1 phase before progressing into apoptosis and pRb may be a negative regulator of this process.

Animals↗
PubMed1996

The interleukin 12 p40 gene promoter is primed by interferon gamma in monocytic cells.

Interleukin (IL) 12 is a proinflammatory cytokine produced by phagocytic cells, B cells, and other antigen-presenting cells that modulates adaptive immune responses by favoring the generation of T helper type 1 cells. IL-12 mediates some of its physiological activities by acting as a potent inducer of interferon (IFN) gamma production by T and natural killer cells. IFN-gamma enhances the ability of the phagocytic cells to produce IL-12 and other proinflammatory cytokines. Thus, IL-12-induced IFN-gamma acts in a positive feedback loop that represents an important amplifying mechanism in the inflammatory response to infections. We show here that IFN-gamma enhances IL-12 production mostly by priming phagocytic cells for lipopolysaccharide (LPS)-induced transcription of the IL-12 p40 gene, which encodes the heavy chain of the IL-12 heterodimer; furthermore, IFN-gamma directly induces transcription of the IL-12 p35 gene, which encodes the light chain of IL-12, and has at least an additive effect with LPS stimulation in inducing its transcription. The priming effect of IFN-gamma on the LPS-induced p40 gene transcription requires preincubation of the cells with IFN-gamma for at least 8 h to obtain a maximal effect. The priming effect of IFN-gamma for IL-12 production is predominantly at the transcriptional level for both the p40 and the p35 gene, and no evidence for a major role of posttranscriptional or translational mechanisms was found. A 3.3-kb human IL-12 p40 promoter construct transfected into cell lines recapitulated the tissue specificity of the endogenous gene, being silent in two human T cell lines, constitutively active in two human Epstein-Barr virus-positive B lymphoblastoid cell lines, and LPS inducible in the human THP-1 and mouse RAW264.7 monocytic cell lines. Because the RAW264.7 cell line is easily transfectable and regulates the endogenous IL-12 p40 gene in response to IFN-gamma or LPS similarly to human monocytes, it was used for analysis of the regulation of the cloned human IL-12 p40 promoter. A requirement for the region between -222 and -204 in both LPS responsiveness and IFN-gamma priming was established. This region contains an ets consensus sequence that was shown to mediate activation of the promoter by IFN-gamma and LPS, as well as by a cotransfected ets-2. The -222 construct was also regulated in a tissue-specific manner. Two other elements, IRF-1 located at -730 to -719, and NF-IL6 at -520 to -512, were also studied by deletion analysis, which did not result in decreased response to IFN-gamma and LPS stimulation.

Animals↗
PubMed1996

Polyenylphosphatidylcholine attenuates non-alcoholic hepatic fibrosis and accelerates its regression.

BACKGROUND/AIMS: Polyenylphosphatidylcholine protects against alcoholic cirrhosis in the baboon. This study assesses whether the antifibrotic effect also pertains to a species other than the baboon and to agents other than alcohol. METHODS: Rats were injected with either CC14 in peanut oil or peanut oil alone, and pair-fed nutritionally adequate liquid diets, with or without polyenylphosphatidylcholine. Other rats were injected with heterologous albumin instead of CC14. To assess whether polyenylphosphatidylcholine is active on established fibrosis, rats were also given CC14 for 8 weeks, and then divided into two groups and pair-fed a diet with or without polyenylphosphatidylcholine. RESULTS: After 8 weeks of CC14, the animals were sacrificed; chromotrope aniline blue and Sirius red stains of liver revealed fibrosis or cirrhosis in animals given CC14 alone, whereas the effect was attenuated in the polyenylphosphatidylcholine-supplemented animals. Hepatic collagen content was decreased by 25 to 32% (p < 0.05) and serum ALT and AST were significantly less increased. The expression of liver collagen type I mRNA was significantly increased in CC14 treated rats and was not significantly affected by polyenylphosphatidylcholine although there was a trend towards a lesser increase polyenylphosphatidylcholine also attenuated liver fibrosis produced by the injection of heterologous albumin. CC14-induced liver fibrosis regressed more rapidly in polyenylphosphatidylcholine-treated animals than controls, both histologically and by measurement of collagen (p < 0.05). CONCLUSIONS: Polyenylphosphatidylcholine (a) attenuates hepatic fibrosis induced by CC14 or human albumin in rats; and (b) accelerates the regression of pre-existing fibrosis.

Animals↗
PubMed1996

Nitric oxide metabolite levels in acute vaso-occlusive sickle-cell crisis.

OBJECTIVES: 1) To measure nitric oxide (NO) metabolite levels in patients presenting to the ED in acute vaso-occlusive sickle-cell crisis (SCC), and 2) to determine whether a relationship exists between NO metabolite levels and pain. METHODS: A prospective, observational study of patients with documented sickle-cell anemia (SCA), aged > or = 18 years, presenting in typical, acute SCC was conducted in an urban, university teaching hospital. Excluded were those with atypical pain or acute, coexistent disease (as evidenced by fever, tachycardia, tachypnea, or hypotension). Pain scores were measured by a 10-cm visual analog scale (VAS). Blood NO metabolite levels for SCC patients and control subjects (healthy volunteers, n = 9; SCA control subjects not in SCC, n = 10) were determined using an NO-specific chemiluminescence technique that measured plasma nitrite and nitrate, the stable end-products of NO. The acute SCC patients were divided into 3 groups, with the range for the SCC-normal (n = 5) group defined as within 2 SD of the healthy volunteer control patients. The SCC-low patients (n = 21) had NO metabolite levels below this range and the SCC-high (n = 21) patients had levels above this range. RESULTS: The SCA and healthy volunteer control groups had similar NO metabolite levels (25.3 vs 22.6 mumol; p = 0.10). The 3 acute SCC groups had the following mean NO levels: 1) SCC-normal = 21.3 +/- 1.6 mumol; 2) SCC-low = 7.2 +/- 1.1 mumol; and 3) SCC-high = 43.7 +/- 3.5 mumol. The SCC-high NO-level group had significantly lower VAS pain scores when compared with the SCC-low and SCC-normal NO-level groups (6.52 +/- 1.85 cm vs 8.76 +/- 0.83 cm, and 8.62 +/- 1.29 cm, p = 0.02). CONCLUSION: NO metabolite levels vary in SCC patients. Elevated levels are associated with lower pain scores, while lower levels are associated with higher pain scores, indicating that NO metabolites may potentially represent a marker for compensatory mechanisms in SCC tissue ischemia. Further work is needed to delineate the usefulness of NO metabolites in assessing the severity of SCC.

Acute Disease↗
PubMed1996

Detection of two clusters of mechanical properties of smooth muscle along the airway tree.

Heterogeneity of function of airway smooth muscle along the airways may be of great importance in regulating regional ventilation and in the pathogenesis of asthma. To investigate the distribution of mechanical properties of airway smooth muscle along the airway, muscle strips free of cartilage and epithelium from the trachea down to bronchial generation 6 were studied by employing electrical field stimulation. Results showed that smooth muscle mechanical performance decreased progressively down the airway tree. Cluster analysis further indicated that smooth muscle from these airways could be divided into two groups: 1) an extrapulmonary group, which contains muscle from the trachea and bronchial generations 1 and 2 and is characterized by higher maximum shortening capacity and zero-load velocity of shortening (V0) in early shortening, the expected decrease of V0 values (the so-called latch phase) in the later phase of shortening, and lower sensitivity to stimulation; and 2) an intrapulmonary group, which contains bronchi from generations 3-6 and has a lower maximum shortening capacity and V0 in early shortening but higher sensitivity to stimulation. The relatively lower mechanical performance of intrapulmonary bronchial smooth muscle may represent a safety device that prevents excessive smooth muscle shortening in vivo.

Animals↗
PubMed1996

Activation of purinergic receptors triggers oscillatory contractions in adult rat ventricular myocytes.

Extracellular ATP is an important neurotransmitter that modulates cardiac function by activation of purinergic receptors. In this study, the effect of P2 purinergic receptor activation on contractions and on [Ca2+]i was investigated in adult rat ventricular myocytes. Fura 2 was used to measure [Ca2+]i, and video edge detection was used to measure contraction. Superfusion of 2-methylthio-adenosine-5'-triphosphate (2-M-S-ATP) over quiescent myocytes induced oscillations in contraction and in [Ca2+]i. The frequency of the oscillatory contractions increased with increasing concentrations of 2-M-S-ATP, but the amplitude of contractions varied from cell to cell and was independent of the concentration of 2-M-S-ATP. During electrical stimulation, activation of purinergic receptors in myocytes potentiated the amplitude of contraction and induced arrhythmias. In populations of quiescent myocytes, the plateau phase of the [Ca2+]i signal evoked by 2-M-S-ATP could be shown to represent summed oscillations in [Ca2+]i in individual cells. Pretreatment of quiescent myocytes with thapsigargin or caffeine reduced or abolished the oscillations in contractions and in [Ca2+]i triggered by 2-M-S-ATP, indicating a dependence of the oscillations on uptake and release of Ca2+ by the sarcoplasmic reticulum. These data demonstrate the novel phenomenon that activation of purinergic receptors in quiescent myocytes stimulates oscillations in [Ca2+]i and contraction. In electrically stimulated myocytes, activation of purinergic receptors triggers oscillatory contractions and potentiates the amplitude of electrically triggered contractions.

Adenosine Triphosphate↗
PubMed1996

The Drosophila morphogenetic protein Bicoid binds DNA cooperatively.

The Drosophila morphogenetic protein Bicoid, encoded by the maternal gene bicoid, is required for the development of the anterior structures in the embryo. Bicoid, a transcriptional activator containing a homeodomain, is distributed in an anterior-to-posterior gradient in the embryo. In response to this gradient, the zygotic gene hunchback is expressed uniformly in the anterior half of the embryo in a nearly all-or-none manner. In this report we demonstrate that a recombinant Bicoid protein binds cooperatively to its sites within a hunchback enhancer element. A less than 4-fold increase in Bicoid concentration is sufficient to achieve an unbound/bound transition in DNA binding. Using various biochemical and genetic methods we further demonstrate that Bicoid molecules can interact with each other. Our results are consistent with previous studies performed in the embryo, and they suggest that one mechanism to achieve a sharp on/off switch of gene expression in response to a morphogenetic gradient is cooperative DNA binding facilitated by protein-protein interaction.

Animals↗
PubMed1996

Observations on the vascular structure of malignant tumours of the mandible using specimen arteriography.

OBJECTIVES: To investigate the vascular structure of different malignant tumours of the mandible. METHODS: Specimen arteriography was performed on seven cases of squamous cell carcinoma of gingiva and four sarcomas. RESULTS: Malignant tumours could be divided into two groups: (1) the hypervascular sarcomas in in which the arteries were dilated, distributed evenly and arranged in a bush-like pattern; and (2) hypovascular squamous cell carcinoma with scanty arteries distributed unevenly without dilatation; in some areas, the shape of arteries was irregular and arranged in a network pattern. CONCLUSIONS: The variation in vascular structure of malignant tumours of the mandible may be related to the outcome of regional chemotherapy.

Adolescent↗
PubMed1996

Cloning of the human erythropoietin exons and their expression in COS-7 cells.

Human erythropoietin exons (hEPO-E) were isolated from a Chinese fetal liver DNA library by using the PCR method with gene recombination. One nucleotide mutation was found in exon 3 leading to the Leu in position 62 being changed to Ser. The hEPO-E coding for the full-length mature protein was inserted into different clone sites of PSV-2-dhfr to create different transferring plasmids (pSV2-dhfr/F1, pSV2/F2, pSV2-dhfr/F3, pSV2-dhfr/F4, pSV2-dhfr/G1, and pSV2-dhfr/G3) which were transfected into COS-7 cells. Results obtained from the comparative experiments indicate that biological activity of hEPO was found in all culture supernatants and its expression level was higher than that of its genome.

Amino Acid Sequence↗
PubMed1996

Chronic suppurative parotitis: a proposed classification.

OBJECTIVE: To propose a practical and reasonable classification of chronic suppurative parotitis (CSP) on the basis of the various entities. MATERIAL AND METHODS: Clinical, laboratory, sialographic, scintigraphic, histopathologic (including ultrastructural) study of recurrent parotid swellings (RPS) was performed in 291 patients over a 10-year period. RESULTS: It is suggested that CSP should be classified into recurrent parotitis in childhood (RPC), recurrent parotitis in adults (RPA), chronic obstructive parotitis (COP) and should be differentiated from other subdivisions including subclinical Sjögren's syndrome (SCSS), chronic parotid swelling of Sjögren's syndrome and sialadenosis with retrograde infection. RPA is a continuation of recurrent parotid swelling from childhood (RPC) to adulthood. Remission can take place spontaneously in RPC and RPA so that self-conservative therapy is mainly used for reducing the parotid swellings. COP is recurrent parotid swellings and/or purulent discharge resulting from various obstructive factors. Mild COP can recover completely with the use of conservative methods, severe COP is often resistant to conservative treatment and should be treated with surgical modality or injection of methyl violet into the diseased gland. Treatment with methyl violet is considered as a convenient and practical method with a definite effect. SCSS is an autoimmune disease and should be treated as systemic disease. CONCLUSIONS: Because there exists confusion in the nomenclature of RPS this revised classification is based on the various entities and can be used as a guide in the diagnosis and treatment of RPS.

Adolescent↗
PubMed1996

[The application of expanded delto-pectoral flaps in the faciocervical region].

The delto-pectoral flap has been used to repair burned scar in the faciocervical region for some years. However, its limited size restricts its application. Furthermore, direct transfer of the flap may result in a swelling and inexpressive face and the donor site needs skin grafting. To avoid the above disadvantages, we have tried pre-expansion of the flap. In this article, the authors report the experiences in the application of the method to eighteen cases, including the surgical procedure, the applied anatomy, and typical cases as well. Also included in the article are the comparison between various therapies to the burned scar of the face, the key points for successful pre-expansion of the delto-pectoral flap, accurately maintaining the desired position. The method has been proved to have many advantages and can be widely applied.

Adolescent↗