A new animal model for Epstein-Barr virus pathogenesis.
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Biomedical subjects
Publications and source records attributed to F Wang.
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ATP was added to the cultured sensory neurons obtained from the dorsal root ganglia of the neonatal rats and PBS was added to serve as control. MTT assays were conducted to evaluate the survival and activity of the cultured neurons. And the silicone regenerative chamber was used after the sciatic nerve incision of the mature SD rat. 1 mmol/L ATP was injected into the left chamber and 0.09% natrium chloride was injected into the right chamber as controls. The changes of nitric oxide synthase (NOS) activity in the corresponding dorsal root ganglia were measured histochemically and image analysis was also performed 4 days after the sciatic nerve injury. The results showed that extracellular ATP could enhance the survival of the neurons and the number of NOS positive neurons were significantly different between the ATP and control groups (P < 0.05). It was suggested that extracellular ATP had neurotrophic effect on neurons survival and could inhibit the NOS activity of the sensory neurons after the peripheral nerve incision, hence exerting the protective effect on the neurons, which was valuable for nerve regeneration after nerve injury.
We report here surveillance data on the bacterial resistance of clinical isolates from 11 Shanghai hospitals in 1999, for guidance in the clinical use of antibacterial agents. Of the 14,855 strains collected, 5130 (34.5%) were Gram-positive cocci and 9725 (65.5%) were Gram-negative bacilli. The most common organisms in descending order of frequency, were: Escherichia coli (16%), coagulase-negative staphylococci (CNS; 14.3%), Klebsiella spp. (12.3%), Staphylococcus aureus (11.5%), Pseudomonas aeruginosa (9.2%), Acinetobacter spp. (8.1%), and Enterococcus spp. (6.6%). Methicillin-resistant strains accounted for 64% and 77% of S. aureus and CNS, respectively. The methicillin-sensitive strains were susceptible to most agents tested, while most methicillin-resistant strains were resistant to these agents. No vancomycin-resistant staphylococci were identified. Vancomycin-resistant strains accounted for 3.6% of Enterococcus fecalis and 1.7% of E. fecium. E. coli strains resistant to piperacillin, gentamicin, and fluoroquinolones accounted for 50% or more of the strains, and the resistance rates of Klebsiella spp., Enterobacter spp., Citrobacter spp., and Acinetobacter spp. to third-generation cephalosporins had increased markedly compared with rates in recent years. Resistance rates of P. aeruginosa to ceftazidime and imipenem (27% and 20%, respectively) had also increased compared with rates in recent years. A national strategy on the limited and prudent use of antibiotics is urgently needed.
BACKGROUND: Patients undergoing coronary endarterectomy during coronary artery bypass grafting (CABG) are at increased risk of perioperative myocardial infarction due to coronary intimal disruption. Data assessing the safety of the antifibrinolytic drug tranexamic acid (TA) in patients undergoing this procedure are lacking. METHODS: From September 1997 to December 1999, 221 patients underwent nonemergency primary CABG with endarterectomy of the right coronary artery alone in 149, the left anterior descending in 35, or both right and left anterior descending in 27. TA was administered intraoperatively to 87 patients (TA group: average total dose 62 +/- 4.4 mg/kg; range 20 to 109 mg/kg), and was not administered to 134 patients (No TA group). RESULTS: The patient characteristics of the 2 groups were similar. In-hospital mortality consisted of 2 patients in the TA group and 4 patients in the No TA group. Perioperative myocardial infarction rates were 2% and 5% in the TA and No TA groups, respectively (p = 0.49). The relative risk for any type of perioperative cardiac ischemic event in the TA group versus the No TA group was 0.77 (95% CI; 0.4, 1.2). Patients in the TA group had a significant reduction in postoperative chest tube drainage (685 versus 894 mL in the TA versus No TA groups, respectively) and in the use of fresh-frozen plasma (p = 0.03). CONCLUSIONS: These results suggest that the clinical effectiveness of tranexamic acid in reducing postoperative blood loss in patients undergoing coronary endarterectomy is not associated with a higher incidence of myocardial ischemia-related complications.
Diagnosis of acute osteomyelitis is often challenging but can be made by plain radiograph, bone scan, or MR imaging. This diagnosis may be more problematic in small bones, in diabetic or immunocompromised patients, those partially treated, post-traumatic, previous surgery, or with pre-existing marrow conditions and associated soft tissue infections. CT is the modality of choice for revealing sequestra and cortical erosions in chronic osteomyelitis. Nonenhanced and enhanced STIR or fat-saturated sequences are essential to reveal the marrow abnormality and its extension for diagnosis of subtle cases with neuropathic or other associated conditions. Combined radionuclide scintigraphy becomes necessary in complicated situations.
Bacterial resistance of Streptococcus pneumoniae against erythromycin and clindamycin and resistance phenotypes of erythromycin-resistant Streptococcus pneumoniae were investigated. The MICs of erythromycin and clindamycin against 345 strains of Streptococcus pneumoniae were tested with agar dilution method; the phenotypes of erythromycin-resistant Streptococcus pneumoniae were detected by double-disk test. One hundred and eighty-three and 171 of 345 (53.0% and 49.6%) of isolates had MICs > or =1 microg/ml for erythromycin and for clindamycin. Among erythromycin-resistant Streptococcus pneumoniae, the percentage of cMLS, iMLS and M phenotype was 90.3% (159/176), 5.7% (10/176) and 4.0% (7/176), respectively. The incidence of erythromycin-resistant Streptococcus pneumoniae is very high in Shanghai. The main phenotype is cMLS.
A personal computer-based on-line monitoring and controlling system was developed for the fermentation of microorganism. The on-line HPLC system for the analysis of glucose and ethanol in the fermentation broth was connected to the fermenter via an auto-sampling equipment, which could perform the pipetting, filtration and dilution of the sample and final injection onto the HPLC through automation based on a programmed procedure. The A/D and D/A interfaces were equipped in order to process the signals from electrodes and from the detector of HPLC, and to direct the feed pumps, the motor of stirrer and gas flow-rate controller. The software that supervised the control of the stirring speed, gas flow-rate, pH value, feed flow-rate of medium, and the on-line measurement of glucose and ethanol concentration was programmed by using Microsoft Visual Basic under Microsoft Windows. The signal for chromatographic peaks from on-line HPLC was well captured and processed using an RC filter and a smoothing algorithm. This monitoring and control system was demonstrated to be effective in the ethanol fermentation of Zymomonas mobilis operated in both batch and fed-batch modes. In addition to substrate and product concentrations determined by on-line HPLC, the biomass concentration in Z. mobilis fermentation could also be on-line estimated by using the pH control and an implemented software sensor. The substrate concentration profile in the fed-back fermentation followed well the set point profile due to the fed-back action of feed flow-rate control.
Core molecular orbital contribution to the electronic structure of N2O isomers has been studied using quantum mechanical density functional theory combined with a plane wave impulse approximation method. Momentum distributions of wave functions for inner shell molecular orbitals of the linear NNO, cyclic and linear NON isomers of N2O are calculated through the (e, 2e) differential cross sections in momentum space. This is possible because this momentum distribution is directly proportional to the modulus squared of the momentum space wave function for the molecular orbital in question. While the momentum distributions of the NNO and cyclic N2O isomers demonstrate strong atomic orbital characteristics in their core space, the outer core molecular orbitals of the linear NON isomer exhibit configuration interactions between them and the valence molecular orbitals. It is suggested that the frozen core approximation breaks down in the prediction of the electronic structure of such an isomer. Core molecular orbital contributions to the electronic structure can alter the order of total energies of the isomers and lead to incorrect conclusions of the stability among the isomers. As a result, full electron calculations should be employed in the study of N2O isomerization.
BACKGROUND: Tissue-specific promoters are becoming increasingly important in light of their effects on gene expression in gene therapy experiments. The regulation of gene expression may be as important as the delivery of the gene itself. OBJECTIVES: To determine the effects of the involucrin (INV), keratin 14 (K14) and cytomegalovirus (CMV) promoters on the expression of the reporter gene beta-galactosidase. METHODS: In vivo, plasmid DNA was introduced to BALB/c mice by gene gun. Skin biopsies were taken after 24 h for histology and beta-galactosidase staining. In tissue culture cells, plasmid DNA was introduced by transient transfection to cell lines 293 (transformed primary human embryonal kidney cells), NIH 3T3 (immortalized mouse fibroblasts) and human keratinocytes. Reporter gene expression was assayed by histochemical staining and chemiluminescence. RESULTS: The K14 and INV promoter constructs showed beta-galactosidase gene expression only in the epidermis, while the CMV promoter showed gene expression in both the dermis and epidermis. In cell culture, the INV and K14 promoter constructs demonstrated significant beta-galactosidase expression in human keratinocytes, but minimal expression in 293 and NIH 3T3 cell types. CMV promoter constructs demonstrated significant expression in all cell types. CONCLUSIONS: Gene expression can be regulated by different promoters both in vivo and in cell culture. Based on the physiological expression of the different promoters, gene expression can be restricted to certain cell types, tissues and skin layers.
Clinically indicated endoscopic examinations of 56 patients with spinal cord injury (SCI) (31 for bleeding) were performed over a 3-year period, of which 3 (6%) showed solitary rectal ulcer syndrome (SRUS). The presentation was rectal bleeding or mucoid discharge. The endoscopic appearance was multiple pseudopolyps and occasional mucosal ulcers extending proximally 8 to 40cm from the anus. Mucosal biopsy specimens showed distorted mucosal glands and displaced smooth muscle fibers wrapping around the glands, the hallmark of SRUS. The affected patients had routinely used suppositories and digital stimulation for bowel care and had been paralyzed 7 to 50 years. None had rectal prolapse. These cases show that SRUS (colitis cystica profunda) can be found among patients with SCI.
UNLABELLED: Carbon disulfide is a well-known occupational hazard in the viscose industry, and studies have shown considerable health effects when workers are exposed to high concentrations of this reagent. At exposure levels below the TLV-TWA(31 mg/m3), findings remain contradictory, probably due to deficient exposure data. The present study tries to identify the occupational hazards and thoroughly assess the exposure levels in a Chinese viscose rayon plant. METHODS: An industrial hygienic field survey and a sampling campaign were carried out, including multi-gas monitoring, on-line measurements, and stationary assessment in the spinning hall as well as personal exposure sampling for spinners (by charcoal tube absorbing and GC-FPD analysis). All data was introduced into Foxpro database, and analyzed by Epi info (6.04) and SPSS. RESULTS: On-line measurement showed that the geometric mean (GM) of carbon disulfide exposure amounted to 12.73 mg/m3 in 'exposure' and 0.08 mg/m3 in 'non-exposure' worksites. These concentrations in the air were related to the subject's activities showing the highest levels when they had to open the shield windows of the spinning machines. Stationary exposure measurements of carbon disulfide in the spinning hall amounted to 23.29 mg/m3 GM (range 5.8-97.94 mg/m3). Personal exposure of spinners was about 17.3 mg/m3 GM. Comparing these methods, the personal exposure sampling could exactly express the exposure levels of the worker's contacting situation. The on-line measurement by multi-gas monitor might also be recommended to the factory as it has its own advantages of rapid and independent assessment, but it under-estimates the exposure level.
We characterized the functional and molecular properties of nicotinic acetylcholine receptors (AChRs) expressed by IMR-32, a human neuroblastoma cell line, and compared them to human alpha3 AChRs expressed in stably transfected human embryonic kidney (HEK) cells. IMR-32 cells, like neurons of autonomic ganglia, have been shown to express alpha3, alpha5, alpha7, beta2, and beta4 AChR subunits. From these subunits, several types of alpha3 AChRs as well as homomeric alpha7 AChRs could be formed. However, as we show, the properties of functional AChRs in these cells overwhelmingly reflect alpha3beta4 AChRs. alpha7 AChR function was not detected, yet we estimate that there are 70% as many surface alpha7 AChRs in IMR-32 when compared with alpha3 AChRs. Agonist potencies (EC(50) values) followed the rank order of 1,1-dimethyl-4-phenylpiperazinium (DMPP; 16+/-1 microM) > nicotine (Nic; 48 +/- 7 microM) > or = cytisine (Cyt; 57 +/- 3 microM) = acetylcholine (ACh; 59 +/- 6 microM). All agonists exhibited efficacies of at least 80% relative to ACh. The currents showed strong inward rectification and desensitized at a rate of 3 s(-1) (300 microM ACh; -60 mV). Assays that used mAbs confirmed the predominance of alpha3- and beta4-containing AChRs in IMR-32 cells. Although 18% of total alpha3 AChRs contained beta2 subunits, no beta2 subunit was detected on the cell surface. Chronic Nic incubation increased the amount of total, but not surface alpha3beta2 AChRs in IMR-32 cells. Nic incubation and reduced culture temperature increased total and surface AChRs in alpha3beta2 transfected HEK cells. Characterization of various alpha3 AChRs expressed in HEK cell lines revealed that the functional properties of the alpha3beta4 cell line best matched those found for IMR-32 cells. The rank order of agonist potencies (EC(50) values) for this line was DMPP (14 +/- 1 microM) = Cyt (18 +/- 1 microM) > Nic (56 +/- 15 microM > ACh (79 +/- 8 microM). The efficacies of both Cyt and DMPP were approximately 80% when compared with ACh and the desensitization rate was 2 s(-1). These data show that even with the potential to express several human nicotinic AChR subtypes, the functional properties of AChRs expressed by IMR-32 are completely attributable to alpha3beta4 AChRs.
BACKGROUND: The Cardiac Anesthesia Risk Evaluation (CARE) score is a simple risk classification for cardiac surgical patients. It is based on clinical judgment and three clinical variables: comorbid conditions categorized as controlled or uncontrolled, surgical complexity, and urgency of the procedure. This study compared the CARE score with the Parsonnet, Tuman, and Tu multifactorial risk indexes for prediction of mortality and morbidity after cardiac surgery. METHODS: In this prospective study, 3,548 cardiac surgical patients from one institution were risk stratified by two investigators using the CARE score and the three tested multifactorial risk indexes. All patients were also given a CARE score by their attending cardiac anesthesiologist. The first 2,000 patients served as a reference group to determine discrimination of each classification with receiver operating characteristic curves. The following 1,548 patients were used to evaluate calibration using the Pearson chi-square goodness-of-fit test. RESULTS: The areas under the receiver operating characteristic curves for mortality and morbidity were 0.801 and 0.721, respectively, with the CARE score rating by the investigators; 0.786 and 0.710, respectively, with the CARE score rating by the attending anesthesiologists (n = 8); 0.808 and 0.726, respectively, with the Parsonnet index; 0.782 and 0.697, respectively, with the Tuman index; 0.770 and 0.724 with the Tu index, respectively. All risk models had acceptable calibration in predicting mortality and morbidity, except for the Parsonnet classification, which failed calibration for morbidity (P = 0.026). CONCLUSIONS: The CARE score performs as well as multifactorial risk indexes for outcome prediction in cardiac surgery. Cardiac anesthesiologists can integrate this score in their practice and predict patient outcome with acceptable accuracy.
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Oxidative stress occurs in animals challenged with bacterial endotoxin and can affect the expression of important host inflammatory genes. However, much less is known about the effects of oxidative stress on responses to gram-negative bacteria. The current study compared the effects of redox imbalance on hepatic responses of mice to Escherichia coli bacteria versus purified endotoxic lipopolysaccharide (LPS). Oxidative stress induced by glutathione depletion virtually eliminated hepatic tumor necrosis factor alpha responses to both E. coli and LPS. Inducible NO synthase (iNOS) and intercellular adhesion molecule-1 (ICAM-1) expression was also markedly inhibited by glutathione depletion in LPS-challenged mice, but was unaffected in E. coli-infected animals. Three findings suggested that gamma interferon (IFN-gamma) production explained the differences between LPS and bacterial challenge. Glutathione depletion completely inhibited the IFN-gamma response to LPS, but only partially inhibited IFN-gamma production in infected mice. Exogenous IFN-gamma restored iNOS and ICAM-1 responses to LPS in stressed mice. Conversely, IFN-gamma-deficient, glutathione-depleted mice showed a marked decrease in iNOS and ICAM-1 expression when challenged with E. coli. These findings indicate that both the nature of the microbial challenge and the production of IFN-gamma can be important in determining the effects of redox imbalance during gram-negative bacterial infections.
The identification of a novel coregulator for nuclear hormone receptors, designated NRIF3, was recently reported (D. Li et al., Mol. Cell. Biol. 19:7191-7202, 1999). Unlike most known coactivators, NRIF3 exhibits a distinct receptor specificity in interacting with and potentiating the activity of only TRs and RXRs but not other examined nuclear receptors. However, the molecular basis underlying such specificity is unclear. In this report, we extended our study of NRIF3-receptor interactions. Our results suggest a bivalent interaction model, where a single NRIF3 molecule utilizes both the C-terminal LXXIL (receptor-interacting domain 1 [RID1]) and the N-terminal LXXLL (RID2) modules to cooperatively interact with TR or RXR (presumably a receptor dimer), with the spacing between RID1 and RID2 playing an important role in influencing the affinity of the interactions. During the course of these studies, we also uncovered an NRIF3-NRIF3 interaction domain. Deletion and mutagenesis analyses mapped the dimerization domain to a region in the middle of NRIF3 (residues 84 to 112), which is predicted to form a coiled-coil structure and contains a putative leucine zipper-like motif. By using Gal4 fusion constructs, we identified an autonomous transactivation domain (AD1) at the C terminus of NRIF3. Somewhat surprisingly, full-length NRIF3 fused to the DNA-binding domain of Gal4 was found to repress transcription of a Gal4 reporter. Further analyses mapped a novel repression domain (RepD1) to a small region at the N-terminal portion of NRIF3 (residues 20 to 50). The NRIF3 gene encodes at least two additional isoforms due to alternative splicing. These two isoforms contain the same RepD1 region as NRIF3. Consistent with this, Gal4 fusions of these two isoforms were also found to repress transcription. Cotransfection of NRIF3 or its two isoforms did not relieve the transrepression function mediated by their corresponding Gal4 fusion proteins, suggesting that the repression involves a mechanism(s) other than the recruitment of a titratable corepressor. Interestingly, a single amino acid residue change of a potential phosphorylation site in RepD1 (Ser(28) to Ala) abolishes its transrepression function, suggesting that the coregulatory property of NRIF3 (or its isoforms) might be subjected to regulation by cellular signaling. Taken together, our results identify NRIF3 as an interesting coregulator that possesses both transactivation and transrepression domains and/or functions. Collectively, the NRIF3 family of coregulators (which includes NRIF3 and its other isoforms) may play dual roles in mediating both positive and negative regulatory effects on gene expression.
PURPOSE: To investigate the feasibility of an automated implementation of a beam equalization technique and to evaluate the experimental performance of the prototype system. MATERIALS AND METHODS: X-ray beam equalization involved the process of low-dose image acquisition, attenuator thickness calculation, mask generation, mask positioning, equalized image acquisition, and mask reshaping. The entire equalization process was performed in approximately 7 seconds. The equalized images were assessed both qualitatively and quantitatively by using a humanoid phantom and a swine animal model. The general image quality was assessed for the ability to visualize arterial branches and other anatomic structures. The level of equalization was quantitatively assessed by segmenting the images into an 8 x 8 matrix of square regions. RESULTS: The ratio of the root-mean-squared variance for the equalized and unequalized images from humanoid phantom and swine animal studies was 0.49 and 0.59, respectively. Furthermore, qualitative assessment of the images showed substantial improvement in image quality and visualization of arterial branches after beam equalization in phantom and animal studies. CONCLUSION: Automated area beam equalization is feasible and improves image quality in previously underpenetrated regions of the image.
Sialyl-Tn (STn), a mucin-associated disaccharide antigen carried by apomucins such as MUC2, plays an important role in tumor biology. However, little is known about the subcellular localization and compartments involved in STn synthesis. In this study we used immunoelectron microscopy to localize STn and MUC2 apomucin in human colorectal tissues. MUC2 apomucin was localized predominantly in the rough endoplasmic reticulum (RER) in normal colorectal mucosa (n=6), colorectal adenoma (n=8), and colorectal adenocarcinoma (n=10). STn, recognized by monoclonal antibody TKH2, was not readily detectable in normal colorectal mucosa but becomes manifest in both trans-Golgi apparatus and mucin droplets in colorectal adenoma. In colorectal adenocarcinoma, STn was localized not only in late but also in early Golgi compartments, and particularly in some RER lumens. Furthermore, electron microscopic in situ hybridization revealed that gold particles representing MUC2 mRNA are primarily localized over the RER. Our findings indicate that in colorectal adenoma STn sialylation takes place in the trans-Golgi apparatus, whereas in colorectal cancer STn sialylation occurs in all the Golgi compartments and in the RER.