Modulation of nitric oxide on lymphokine-activated killer cells in patients with bladder cancer.
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Cell Line, Tumor↗
Biomedical subjects
S Fu
Publications and source records attributed to S Fu.
Deficient iNOS in inflammatory bowel disease intestinal microvascular endothelial cells results in increased leukocyte adhesion.
Microvascular endothelial cells play a key role in inflammation by undergoing activation and recruiting circulating immune cells into tissues and foci of inflammation, an early and rate-limiting step in the inflammatory process. We have previously [Binion et al., Gastroenterology112:1898-1907, 1997] shown that human intestinal microvascular endothelial cells (HIMEC) isolated from surgically resected inflammatory bowel disease (IBD) patient tissue demonstrate significantly increased leukocyte binding in vitro compared to normal HIMEC. Our studies [Binion et al., Am. J. Physiol.275 (Gastrointest. Liver Physiol. 38):G592-G603, 1998] have also demonstrated that nitric oxide (NO) production by inducible nitric oxide synthase (iNOS) normally plays a key role in downregulating HIMEC activation and leukocyte adhesion. Using primary cultures of HIMEC derived from normal and IBD patient tissues, we sought to determine whether alterations in iNOS-derived NO production underlies leukocyte hyperadhesion in IBD. Both nonselective (N(G)-monomethyl-L-arginine) and specific (N-Iminoethyl-L-lysine) inhibitors of iNOS significantly increased leukocyte binding by normal HIMEC activated with cytokines and lipopolysaccharide (LPS), but had no effect on leukocyte adhesion by similarly activated IBD HIMEC. When compared to normal HIMEC, IBD endothelial cells had significantly decreased levels of iNOS mRNA, protein, and NO production following activation. Addition of exogenous NO by co-culture with normal HIMEC or by pharmacologic delivery with the long-acting NO donor detaNONOate restored a normal leukocyte binding pattern in the IBD HIMEC. These data suggest that loss of iNOS expression is a feature of chronically inflamed microvascular endothelial cells, which leads to enhanced leukocyte binding, potentially contributing to chronic, destructive inflammation in IBD.
Cell Adhesion↗
Reactions of hypochlorous acid with tyrosine and peptidyl-tyrosyl residues give dichlorinated and aldehydic products in addition to 3-chlorotyrosine.
The toxicity of hypochlorous acid (HOCl) generated from activated neutrophils has been associated with several pathological processes such as atherosclerosis. Formation of 3-chlorotyrosine (Cl-Tyr) has been used as a marker for assessing the involvement of HOCl in such processes. In this study, we aimed to investigate the formation of Cl-Tyr from reaction of HOCl with tyrosine (both free and peptide-bound) and the fate of Cl-Tyr under such conditions. Tyrosine, N-acetyltyrosine, bovine serum albumin, and human low density lipoproteins were incubated with a range of reagent hypochlorite concentrations for varying periods in 10 mM phosphate buffer (pH 7.4) at 22 degrees C. The reaction products, and several biological samples, were hydrolyzed (in the case of proteins), isolated, and purified by high pressure liquid chromatography and characterized or quantitated by mass spectrometry and NMR. A significant amount of 3,5-dichlorotyrosine (diCl-Tyr) was obtained from the bovine serum albumin, low density lipoprotein, and some biological samples, in addition to Cl-Tyr, indicating that Cl-Tyr competes effectively for HOCl even when tyrosine is present in great excess. Cl-Tyr and diCl-Tyr were also formed from free tyrosine but then reacted further with HOCl. This finding differs from a claim in the literature that Cl-Tyr was not formed in such a system. The further reaction products of Cl-Tyr and diCl-Tyr with HOCl were elucidated as their corresponding mono- and dichlorinated 4-hydroxyphenylacetaldehydes. These results indicate the importance of assessing other products of HOCl action in addition to Cl-Tyr.
Animals↗
BP1, a new homeobox gene, is frequently expressed in acute leukemias.
Aberrant expression of homeobox genes has been described in primary leukemia blasts. We recently cloned a new cDNA, BP1, which is a member of the homeobox gene family. BP1 expression was investigated in bone marrow samples from acute myeloid leukemia (AML), acute T cell lymphocytic leukemia (ALL) and pre-B cell ALL. Expression levels of two apparent isoforms of BP1, DLX7 and DLX4, were measured in the same samples. They are weakly if at all detectable in normal bone marrow, PHA-stimulated T cells or B cells. BP1 RNA was highly expressed in 63% of AML cases, including 81% of the pediatric and 47% of the adult cases, and in 32% of T-ALL cases, but was not found in any of the pre-B ALL cases. Coexpression of BP1, DLX7 and DLX4 occurred in a significant number of leukemias. Our data, including co-expression of BP1 with c-myb and GATA-1, markers of early progenitors, suggest that BP1 expression occurs in primitive cells in AML. Analysis of CD34+ and CD34- normal bone marrow cells revealed BP1 is expressed in CD34- cells and virtually extinguished in CD34+ cells. Ectopic expression of BP1 in the leukemia cell line K562 increased clonogenicity, consistent with a role for BP1 in leukemogenesis. The presence of BP1 RNA in leukemic blasts may therefore be a molecular marker for primitive cells and/or may indicate that BP1 is an important upstream factor in an oncogenic pathway.
Acute Disease↗
Mobilization of hematopoietic stem cells.
Hematopoietic stem cell transplantation has been extensively exploited as a therapeutic and research modality and has revolutionized current patient care. At present, more and more medical centers use peripheral blood progenitor cells for transplantation by mobilizing hematopoietic stem cells from bone marrow to peripheral blood because of potential advantages of peripheral blood stem cell transplantation over bone-marrow transplantation. Different effective mobilization regimens have been developed recently with chemotherapeutic agents, hematopoietic growth factors or their combination. This article reviews current developments related to hematopoietic stem cell mobilization including the biology of hematopoietic stem cells, strategies for mobilization, management for mobilization failure, mechanisms of mobilization, and side effects during mobilization. Finally, the Initiation-Amplification-Emigration-Adaptation Model is proposed to help aid understanding of the mechanisms of hematopoietic stem cell mobilization and to stimulate development of novel and optimal mobilization strategies for patient care.
Hematopoietic Stem Cell Mobilization↗
Effects of basic fibroblast growth factor (bFGF) on early stages of tendon healing: a rat patellar tendon model.
We studied the effects of basic fibroblast growth factor (bFGF) on cell proliferation, type III collagen expression, ultimate stress and the pyridinoline content in the early stages of healing in rat patellar tendon. 96 male Sprague Dawley rats were injected with increasing doses of basic fibroblast growth factor (bFGF) at 3 days after a "window defect" was induced in the mid-part of the patellar tendon. They were killed at 7 and 14 days after the injury. A dose-dependent increase in the number of proliferating cells and the level of expression of type III collagen was demonstrated at only 7 days post-injury. On the other hand, we found no effects of bFGF on ultimate stress and the pyridinoline content of healing tendons. Only time significantly affected both strength-associated parameters. We showed that in vivo supplementation with bFGF affected the initial events of healing such as cell proliferation and type III collagen expression.
Amino Acids↗
Distribution of a common methylenetetrahydrofolate mutation in six Chinese population groups.
The distribution of the C677T polymorphism was analyzed by the PCR-RFLP technique in the Northeast Han, the Oroqen, the Ewenki, the Daur as well as in Koreans and Mongolians. The results were compared with each other. They revealed that the frequencies of the T allele are quite different (17-47%) among the tested groups and are much higher in Chinese population groups than in others.
Asian People↗
Analysis of DYS19 and DYS287 polymorphisms in the Han population and three other ethnic groups of northeast China.
The allelic distribution of the Y-chromosome specific microsatellite DYS19 in the Han population and in the Daur, Oroqen and Ewenki ethnic groups (Northeast China) was analyzed by PCR and denatured polyacrylamide gel electrophoresis. The allelic distribution in the Han population group is as follows: A = 2.90%, B = 26.09%, C = 26.09%, D = 29.98%, E = 15.94%. This allelic distribution differs statistically significant from that observed in the three other ethnic groups (p < 0.05). Furthermore the polymorphism of the Y-chromosome specific Alu insert sequence DYS287 was tested in these four groups. However, no Alu sequence insert was found.
Alleles↗
Analysis of polymorphic mitochondrial DNA in the Han population and three other ethnic groups of northeast China.
By means of the PCR-RFLP technique the polymorphism of a 465 fragment of mtDNA D loop was analyzed in 36 individuals of the Northeast Han, 30 individuals of the Daur, 32 individuals of the Oroqen and 30 individuals of the Ewenki. All these individuals were randomly selected and are healthy. 27 restriction patterns were obtained by assaying the RFLP fragments of the mtDNA D loop. The mean nucleid acid substitute frequencies at each site are the following: Northeast Han = 0.0291, Oroqen = 0.0477, Ewenki = 0.0563, Daur = 0.0541. The genetic relationship between these four populations was analyzed by the UPG cluster analysis.
Asian People↗
The therapeutic effect of Ginkgo biloba extract in SHR rats and its possible mechanisms based on cerebral microvascular flow and vasomotion.
This paper aimed to investigate the therapeutic effect of an extract of Ginkgo biloba leaves (EGb 761) on hypertension and its possible mechanisms in the view of cerebral microcirculation. Twenty normotensive rats and 24 SHR rats were used. Surgical preparation was made to produce a cranial window for observation of the capillary network on the cerebral cortex. The intravital videomicroscopy equipped with digital image processing system and laser Doppler flowmeter were used for this study. The arterial blood pressure, red cell velocity (V), microvacular diameter (D), number of open capillaries (OCN), circulating endothelial cells (CEC) in blood, relative blood flow (Flow) and frequency (Fc), amplitude (AMP) of vasomotion were measured. The obtained data were compared between EGb-treated rats that received per os 100 mg/kg/d for 9 days and placebo control rats. Untreated SHR rats showed very severe dysfunction in the microcirculation with high blood pressure (213 +/- 16.7 mmHg). The blood pressure decreased significantly to 153 +/- 20 mmHg in EGb-treated SHRs group, compared with those of untreated rats (p < 0.01). Both normotensive and hypertensive rats increased the blood flow velocity and LDF flow after EGb-treatment. The vasomotion property, the CEC and OCN changed greatly in EGb-treated SHR rats, but no significant difference was observed in normotensive rats. It was suggested that EGb 761 had therapeutic effect on SHR rats by increasing blood perfusion, regulating vasomotion function, opening efficiently capillaries and releasing the peripheral resistance. The injured vascular endothelium of SHR rats was also partly reversed by EGb-treatment. It was concluded that EGb 761 could be used to regulate hypertension and to protect the cerebral microcirculatory function.
Animals↗
[Retrovirus vector expresses luciferase and TPO gene under the control of doxycycline in NIH3T3 cells].
OBJECTIVE: To use the retrovirus vector Tet-On regulation system to control the expression of TPO and luciferase gene by doxycycline (Dox). METHODS: Packing the recombinant Tet-On retrovirus, and infecting it to NIH3T3 cells to establish recombined Tet-On retrovirus cell lines, RevTet-On3T3. To construct the pRevTRETPO, the new cell line was transferred to PT67 cell to package the recombinant retrovirus which contained both the TRE and TPO. This virus was RevTet-On3T3 used to establish the two inserting recombinant retrovirus cell strain RevTet-On3T3TPO. Doxycycline was used to control the expression of TPO in RevTet-On3T3. RESULTS: When Dox (5 mg/L) was added to the RevTet-On3T3 cells, a higher activity of luciferase was seen (mean, 5.9 x 10(4) RLU.S) compared with the mean activity (2.6 x 10(3) RLU.S) in the absence of Dox. When Dox was added to the RevTet-On3T3TPO cells, cell populations had high TPO expression (mean, 12.8 ng/ml) in 5 mg/L of Dox (On state), and a low TPO expression (mean, 0.56 ng/ml) in the absence of Dox (Off state). CONCLUSION: The RevTet-On gene expression system could modulate the expression of multiple genes by tetracycline and its derivatives, providing a useful tool for gene therapy.
3T3 Cells↗
[Relationship between light and regeneration character of Huangshan pine].
The growth and physiological characters of 1-3 years old Huangshan pine seedlings in pure and mixture plots with four different transparancy were determined, and the germination percentage of seeds and their complex stressing vigour were tested. Meanwhile, the regeneration mechanism of Huangshan pine in natural forest was studied in this paper. The results showed that the plumpness of seeds was 65%, the germination percentage was 27%, and the complex stressing vigour was 26%. The lower germination percentage affected the reproduction of Huangshan pine. The photosynthetic rate was more affected by solar energy, but less by humidity and surface temperature. With high light composation, Huangshan pine's regeneration under forest was difficult. The seedlings of Huangshan pine in mixed coniferous and broadleave tree plots were at a disadvantage because of slow growth and insufficient light.
Ecosystem↗
[Effect of effective fraction of Radix Salviae Miltiorrhizae on procollagen gene expression in fracture healing].
OBJECTIVE: To investigate effect of effective fraction of Radix Salviae Miltiorrhizae (RS9403) on procollagens and transforming growth factor beta 1(TGF beta 1) gene expression in fracture callus, and to explore the mechanism of RS9403 in enhancing fracture healing. METHODS: Standardized radial fractures were performed in 24 Wistar rats, which were randomized into three groups: the RS9403 group, the Sanhua Jiegu powder (SHJGP) group and the blank control model group. They were fed orally with RS9403, SHJGP and normal saline respectively, and were sacrificed on the 3rd day, 1st week, 2nd week and 4th week after fracture in batches. In situ localization of procollagens and TGF beta 1 gene expression were examined on the cryosection of rat fracture callus. RESULTS: On the 3rd day after fracture, the TGF beta 1 gene expression at the site of fracture in the RS9403 group and the SHJGP group was significantly higher than that in the blank control group, and type-III procollagen gene expression was observed in the two groups. At the end of 1st week after fracture, the pro alpha 1(III) expression in fibroblast and chondrocyte-like cells was dominant. The local type II and I procollagen gene expressions in the RS9403 and the SHJGP group were enhanced simultaneously with TGF beta 1 gene expression compared with those in the blank control group. At the end of 2nd week the RS9403 and the SHJGP group were characterized by a marked increase in the mRNA levels of type I procollagen, the numbers of hypertrophic chondrocytes was larger than that in the control group, and the type II procollagen expression declined markedly. The shared phenotype expression was confirmed at this stage, especially in the RS9403 and the SHJGP group. At the end of 4th week, the cartilagi nous callus was almost all replaced by the bone tissue. In the RS9403 and the SHJGP group, the replacement was nearly complete, few type I procollagen mRNA positive osteoblasts and hypertrophic chondrocytes were found scattering in the woven bone and remnants of cartilaginous callus. CONCLUSION: RS9403 could affect the procollagens expression to enhance the healing of bone fracture as SHJGP do, maybe, by modulating the TGF beta 1 expression.
Animals↗
[Effects of B-ultrasonic radiation during pregnancy on reproductive outcome].
OBJECTIVE: To study the effects of B-ultrasonic scanning during pregnancy on reproductive outcome, including pregnant outcome and neonatal status. METHODS: Data of reproductive outcome and B-ultrasonic examination were extracted from the questionnaires and obstetric records of 12,224 primigravidas in eight hospitals of Beijing during May 1992 to April 1993. Their reproductive outcomes were analyzed according to frequency and gestational weeks they had B-ultrasonic examination taken, adjusted for potential confounding factors, such as age at pregnancy, smoking and drinking during pregnant period and history of spontaneous abortion, by logistic regression. RESULTS: There was no statistically significant difference in reproductive outcomes between groups, who had B-ultrasonic examination taken for two, three and four times or more and who had only once, respectively, who had more than four times and who had less than three times, and who had their first examination before and after the eighth week of gestation, or before and after the 12th week of gestation. CONCLUSION: No adverse reproductive outcome due to exposure to diagnostic dose of B-ultrasound radiation during pregnancy has been found.
Abortion, Spontaneous↗
[Toxicity to neural cell development of lead and its relation to glutathione].
OBJECTIVE: To investigate the toxicity of lead to embryonic neural cell development and its relation to glutathione level. METHODS: Rat midbrain micromass culture method was used to observe effects of lead on viability, differentiation and glutathione (GSH) content of embryonic neural cells. RESULTS: Lead concentrations at 3.69 micromol/L and 1.03 micromol/L could inhibit neural cell variability and differentiation by 50%, respectively, both in a significant dose-response pattern. Lead also could cause increase level of glutathione. N-acetylcysteine (NAC), an antioxidant, could not only reduce adverse effect of lead on GSH, but also antagonize its toxicity to cell survival and differentiation. CONCLUSION: Lead can specifically inhibit neural cell differentiation and its neurotoxic effects on brain cell development correlated to imbalance in redox status which is mainly mediated by decrease of GSH content.
Acetylcysteine↗
Increased expression and cellular localization of inducible nitric oxide synthase and cyclooxygenase 2 in Helicobacter pylori gastritis.
BACKGROUND & AIMS: Inducible nitric oxide synthase (iNOS) and cyclooxygenase (COX)-2 are important regulators of mucosal inflammation and epithelial cell growth. To determine the role of iNOS and COX-2 in Helicobacter pylori-induced tissue injury, we compared their gene expression in H. pylori-induced gastritis with that in normal gastric mucosa and in non-H. pylori gastritis. METHODS: In 43 patients, we assessed H. pylori infection status, histopathology, messenger RNA (mRNA) and protein expression, and cellular localization of iNOS and COX-2. RESULTS: By reverse-transcription polymerase chain reaction (RT-PCR), antral iNOS and COX-2 mRNA expression was absent to low in normal mucosa (n = 10), significantly increased in H. pylori-negative gastritis (n = 13), and even more markedly increased in H. pylori-positive gastritis (n = 20). Increased iNOS and COX-2 levels were confirmed by Northern and Western blot analysis and were both greater in the gastric antrum than in the gastric body of infected patients. Immunohistochemistry also showed increased expression of both genes in H. pylori gastritis: iNOS protein was detected in epithelium, endothelium, and lamina propria inflammatory cells, and COX-2 protein localized to mononuclear and fibroblast cells in the lamina propria. CONCLUSIONS: iNOS and COX-2 are induced in H. pylori-positive gastritis and thus may modulate the inflammation and alterations in epithelial cell growth that occur in this disease. Higher levels of iNOS and COX-2 in H. pylori-positive vs. -negative gastritis and in gastric antrum, where bacterial density is greatest, suggest that expression of these genes is a direct response to H. pylori infection.
Cyclooxygenase 2↗
The voltammetric behavior of bone marrow of leukaemia and its clinical application.
The electrochemical voltammetric behavior of bone marrow of leukaemia has been investigated by a self-devised cytosensing system. The two anodic peaks of erythrocytes (red blood cells, RBC) in bone marrow of leukaemia appeared at 0.73 +/- 0.03 and 0.83 +/- 0.02V vs. SCE, respectively, on the first scan. The anodic peak of leukocytes (white blood cells, WBC) appeared at 0.32 +/- 0.03V vs. SCE. The anodic peak of RBC at 0.83V disappeared when the patients were cured. The experimental results show that the voltammetric behavior of erythrocytes is in constant contact with the initial stage of leukaemia. The cyclic voltammetric behavior of 40 cases of leukaemia including acute myeloid leukaemia (AML) and chronic myeloid leukaemia (CML) and 10 cases of healthy volunteer peripheral blood was studied. The cyclic voltammetric behavior of erythrocytes may provide a simple and specific marker for diagnosis of leukaemia.
Bone Marrow↗
Stable markers of oxidant damage to proteins and their application in the study of human disease.
The mechanisms of formation and the nature of the altered amino acid side chains formed on proteins subjected to oxidant attack are reviewed. The use of stable products of protein side chain oxidation as potential markers for assessing oxidative damage in vivo in humans is discussed. The methods developed in the authors laboratories are outlined, and the advantages and disadvantages of these techniques compared with other methodologies for assessing oxidative damage to proteins and other macromolecules. Evidence is presented to show that protein oxidation products are sensitive markers of oxidative damage, that the pattern of products detected may yield information as to the nature of the original oxidative insult, and that the levels of oxidized side-chains can, in certain circumstances, be much higher than those of other markers of oxidation such as lipid hydroperoxides.
Amino Acids↗