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Biomedical subjects

Jianguo Wu

Publications and source records attributed to Jianguo Wu.

30 records · Page 2Linked to original sources

Type IVB piliated Salmonella typhi enhance IL-6 and NF-kappaB production in human monocytic THP-1 cells through activation of protein kinase C.

Salmonella typhi is an important human pathogen responsible for typhoid fever. Type IVB pili, encoded by the S. typhi pil operon located in the major pathogenicity island, are used to facilitate bacterial entry into human intestinal cells in vitro and may be important in the mediation of enteric fever in humans. However, possible involvement of the type IVB pili of S. typhi in signal transduction in infected immune cells has not been examined previously. In this study, we have compared the effect of piliated and nonpiliated S. typhi on the activities of protein kinase C (PKC), the production of interleukin-6 (IL-6) and nuclear transcription factor NF-kappaB in human monocytic THP-1 cells. We find that piliated S. typhi can stimulate significantly higher activities of PKC, the production of IL-6 and NF-kappaB than a nonpiliated strain based on substrate phosphorolysis kinase assay, Western blot, RT-PCR, and luciferase reporter gene assay. In time course experiments, PKC activity increased in a time-dependent fashion after stimulation by the piliated bacteria. The PKC inhibitor Dequalinium chloride (DECA) remarkably reduced the production of IL-6, NF-kappaB and the activity of PKC induced by the piliated S. typhi. These results suggest that the induction of IL-6 and NF-kappaB depend on the PKC signal pathway. Our report demonstrates that the type IVB pili of S. typhi play important roles in the production of NF-kappaB and the proinflammatory cytokine IL-6, and in the stimulation of PKC activity and therefore, may have effects on the development of fever and other inflammatory responses.

Adult↗

Aptamers that preferentially bind type IVB pili and inhibit human monocytic-cell invasion by Salmonella enterica serovar typhi.

Salmonella enterica serovar Typhi is an important pathogen exclusively for humans and causes typhoid or enteric fever. It has been shown that type IVB pili, encoded by the S. enterica serovar Typhi pil operon located in Salmonella pathogenicity island 7, are important in the pathogenic process. In this study, by using both an adhesion-invasion assay and fluorescence quantitative PCR analysis, we demonstrated that the entry of type IVB piliated S. enterica serovar Typhi A21-6 (pil(+) Km(r)) into human THP-1 monocytic cells was greater than that of a nonpiliated S. enterica serovar Typhi pilS::Km(r) (pil mutant) strain. We have applied a systematic evolution of ligands by exponential enrichment approach to select oligonucleotides (aptamers) as ligands that specifically bind to type IVB pili. Using this approach, we identified a high-affinity single-stranded RNA aptamer (S-PS(8.4)) as a type IVB pilus-specific ligand and further found that the selected aptamer (S-PS(8.4)) could significantly inhibit the entry of the piliated strain (but not that of the nonpiliated strain) into human THP-1 cells. The binding affinities between aptamers and pre-PilS (structural protein of type IVB pili) were determined by nitrocellulose filter-binding assays, and the K(d) value was determined to be 8.56 nM for the S-PS(8.4) aptamer alone. As an example of an aptamer against type IVB pili of S. enterica serovar Typhi, the aptamer S-PS(8.4) can serve as a tool for analysis of bacterial type IVB pilus-host cell interactions and may yield information for the development of putative new drugs against S. enterica serovar Typhi bacterial infections, useful both in prevention of infection and in therapeutic treatment.

Bacterial Adhesion↗

PCR analysis of egyptian respiratory adenovirus isolates, including identification of species, serotypes, and coinfections.

Eighty-eight adenovirus (Ad) isolates and associated clinical data were collected from walk-in patients with influenza-like illness in Egypt during routine influenza surveillance from 1999 through 2002. Respiratory Ad distributions are geographically variable, and serotype prevalence has not been previously characterized in this region. Serotype identity is clinically relevant because it predicts vaccine efficacy and correlates strongly with both clinical presentation and epidemiological pattern. Species and serotype identities were determined using several well-validated multiplex PCR protocols culled from the literature and supplemented with a few novel primer sets designed to identify rare types. The isolates included common species B1 serotypes (Ad3 and Ad7), common species C serotypes (Ad1, Ad2, and Ad5), the less common species B2 serotype Ad11, and three isolates of the rare species B1 serotype Ad16. Two isolates that appear to be variant Ad16 were also identified. Fifteen coinfections of multiple adenoviral types, primarily AdB/AdC and Ad3/Ad7 dual infections, were detected. The majority of these were verified using redundant PCR tests targeted at multiple genes. PCR is able to resolve coinfections, in contrast to traditional serum neutralization tests. PCR is also comparatively rapid and requires very little equipment. Application of the method allowed an inclusive determination of the serotypes found in the Egyptian respiratory sample set and demonstrated that coinfections are common and may play a previously unrecognized role in adenovirus pathogenesis, evolution, and epidemiology. In particular, coinfections may influence adenoviral evolution, as interserotypic recombination has been identified as a source of emerging strains.

Adenovirus Infections, Human↗

Ecosystem stability and compensatory effects in the Inner Mongolia grassland.

Numerous studies have suggested that biodiversity reduces variability in ecosystem productivity through compensatory effects; that is, a species increases in its abundance in response to the reduction of another in a fluctuating environment. But this view has been challenged on several grounds. Because most studies have been based on artificially constructed grasslands with short duration, long-term studies of natural ecosystems are needed. On the basis of a 24-year study of the Inner Mongolia grassland, here we present three key findings. First, that January-July precipitation is the primary climatic factor causing fluctuations in community biomass production; second, that ecosystem stability (conversely related to variability in community biomass production) increases progressively along the hierarchy of organizational levels (that is, from species to functional group to whole community); and finally, that the community-level stability seems to arise from compensatory interactions among major components at both species and functional group levels. From a hierarchical perspective, our results corroborate some previous findings of compensatory effects. Undisturbed mature steppe ecosystems seem to culminate with high biodiversity, productivity and ecosystem stability concurrently. Because these relationships are correlational, further studies are necessary to verify the causation among these factors. Our study provides new insights for better management and restoration of the rapidly degrading Inner Mongolia grassland.

Biodiversity↗

Specific inhibition effects of N-pentafluorobenzyl-1-deoxynojirimycin on human CD4+ T cells.

We first synthesized N-pentafluorobenzyl-1-deoxynojirimycin (5F-DNM), one new derivative of 1-deoxynojirimycin (DNM). Effects on human peripheral blood mononuclear cells (PMBC) and secretion of cytokines from human PBMC by 5F-DNM were investigated. It was first found that 5F-DNM remarkably inhibited the secretion of interleukin-4 (IL-4) and had a specific inhibition on the expression of CD4 molecules. 5F-DNM, much less toxic than cyclosporin A, might be used as a new candidate of immunosuppressant for specifically treating Th2-mediated immune diseases.

1-Deoxynojirimycin↗

Molecular analysis of adenovirus isolates from vaccinated and unvaccinated young adults.

Infections of adenovirus type 4 (Ad4) and Ad7 were discovered among previously vaccinated individuals through febrile respiratory illness surveillance at military recruit camps. Genetic analysis was performed on these isolates and a sample of adenovirus isolates from unvaccinated patients. Antigenic regions of the adenovirus hexon gene from 21 vaccinated and 31 unvaccinated patients were sequenced and compared to homologous regions of Ad4 and Ad7 vaccine strains and of other representative hexon sequences archived in GenBank. The phylogenetic distribution of sequences from vaccinated individuals closely resembled those from unvaccinated individuals. The most common Ad7 strain was the Ad7d2 hexon genotype, and the most common Ad4 strain was a genotype nearly identical to the recently discovered Z-G 95-873 Ad4 variant. Near exclusive isolation of Ad4 since 1999 indicates that the Ad4 variant is currently responsible for the vast majority of adenovirus morbidity in military recruit camps. Different ratios of nonsynonymous to synonymous nucleotide substitution rates in known antigenic regions compared to nonantigenic regions indicated positive selection for diversity in the antigenic regions and purifying selection in the nonantigenic regions.

Adenovirus Infections, Human↗

[Impact of land-use change on soil carbon storage].

Through comparing the concentration and inventory of soil organic carbon (SOC) and its distribution in soil profiles under cropland, rangeland, natural secondary forest (brushwood, natural secondary forest dominated by Querces liaotungensis or Populus davidiana) and larch plantations (13, 18 and 25 years old Larix principisrupprechtil), this paper studied the effect of land use change from natural secondary forest to cropland or rangeland as well as from cropland or rangeland to plantation on SOC storage in the Liupan mountain forest zone. The results showed that the concentration of SOC in 0-110 cm soil layer under cropland and rangeland was 54% and 27% lower than that under natural secondary forest, respectively. The difference of SOC concentration between natural secondary forest and cropland or rangeland was greater in 0-50 cm than in 50-110 cm soil layer, while that between larch plantations and cropland or rangeland was greater in 0-40 cm than in 40-110 cm soil layer. The inventory of SOC in 0-110 cm soil layer under cropland and rangeland was respectively 35% and 14% lower than that under natural secondary forest, while 23% lower under cropland and 4% higher under rangeland than that under larch plantations. The difference of SOC inventory between natural secondary forest and cropland or rangeland was greater in 0-50 cm than in 50-110 cm soil layer, while that between plantations and cropland or rangeland was greater in 0-30 cm than in 30-110 cm soil layer. The decreasing magnitude of SOC storage with soil profile depth under natural secondary forest or larch plantations was greater than that under cropland or rangeland. The above-mentioned facts resulted from the changes of SOC input or output and the distribution of roots in soil. The results indicated that the SOC concentration and inventory would decline (mainly in 0-50 cm soil layer) after converting from natural secondary forest to cropland or rangeland, but increase (mainly in 0-30 cm soil layer) following afforestation on cropland. The SOC concentration would increase but its inventory would not change following afforestation on rangeland, and the distribution of the SOC concentration or inventory in soil profile would change with the change of land use in Liupan mountain forest zone.

Agriculture↗

Ecology. Three-Gorges Dam--experiment in habitat fragmentation?

Habitat fragmentation is the primary cause of the loss of biodiversity and ecosystem services, but its underlying processes and mechanisms remain poorly understood. Studies of islands and insular terrestrial habitats are essential for improving our understanding of habitat fragmentation. We argue that the Three-Gorges Dam, the largest that humans have ever created, presents a unique grand-scale natural experiment that allows ecologists to address a range of critical questions concerning the theory and practice of biodiversity conservation.

Animals↗

[The temporal variations of soil respiration under different land use in Liupan Mountain forest zone].

The temporal variations of soil respiration under cropland, rangeland, natural secondary forest (brushwood, nature secondary forest dominated by Querces liaotungensis koiz or Populus davidiana dode) and the plantation of larch (13, 18 and 25-year-old Larix principis-rupprechtil mayr) in Liupan Mountain forest zone was studied. It was found that the rate of soil respiration increased with increasing soil temperature during diurnal variation and the highest temperature was from 13:00 o'clock to 15:00 o'clock, the lowest temperature was from 04:00 o'clock and 08:00 o'clock, the variation of soil respiration rate appeared same trend. The daily rate of soil respiration increased from May to October and which was the highest from August to September and declined on October. The diurnal or seasonal variation of soil respiration rate mainly consisted with the diurnal or seasonal variation of soil temperature and its range under cropland or rangeland was greater than which under forestland (natural secondary forest, plantation or brushwood). The highest rate of soil respiration for diurnal or seasonal variation of soil CO2 efflux under cropland or rangeland was higher than which under the forestland, while the lowest rate of soil respiration under cropland or rangeland was lowered than which under forestland. Annual rate of respiration for natural secondary forest, cropland, rangeland and plantation was 3.96-4.51 t/(hm2.a), 1.91 t/(hm2.a), 5.08 t/(hm2.a) and 4.11-5.55 t/(hm2.a) respectively. The results shows that the diurnal or seasonal variation range of soil respiration rate would increase with conversion from natural forests to cropland or rangeland, while which would decrease following afforestation under cropland or rangeland. In addition, the annual respiration rate would also change with change in land use.

Carbon Dioxide↗

Analysis of differential gene expression by ligation specificity-based transcript profiling.

An open architecture mRNA profiling technology, LEAD (Ligation specificity-based Expression Analysis Display), was developed for studying differential gene expression. This method utilizes restriction enzymes with N(m) degeneracy in their recognition/cleavage sequences to fractionate cDNA population (N represents any one of the four possible bases; while m > or = 1, represents the number of degenerate bases). The fractionated cDNAs are subpooled by selective ligation with specific adapters, and then amplified and labeled by PCR. Fluorescent-labeled cDNA fingerprints are separated by electrophoresis as distinct bands with unique size and sequence, and quantified electronically by using the LEAD Finder program. The specificity of ligation and the uniform efficiency of PCR reaction allow precise quantification of differential gene expression among samples. Transcripts of low abundance (1/100,000 copies) can be detected, allowing the status of nearly all mRNA to be monitored. Because of its sequence independence, this technology can be used to monitor gene expression in both model and nonmodel systems lacking whole genome information. It can also be applied to separate and collect different cDNA species fingerprints to build a nonredundant EST library for microarray and other applications.

Animals↗

Islet transplantation in the twenty-first century.

Isolated islet transplantation is poised for clinical application to treat insulin-dependent diabetes. Unlike exogenous insulin therapy, islet transplantation has promise for preventing and/or reversing the dismal secondary complications of diabetes. Islet transplants are arguably the most unique type of allografts, and we discuss their properties, limitations, and potential in this overview. The induction of immunologic tolerance to allow islet grafts to endure and prevail, without the hardship of chronic immunosuppressive therapy, is a major goal in this field. In this context, we discuss our successful results in preclinical models of primate allogeneic and xenogeneic islet graft tolerance.

Animals↗

The immune decision toward allograft tolerance in non-human primates requires early inhibition of innate immunity and induction of immune regulation.

Brief treatment of rhesus macaques with immunotoxin plus 15-deoxyspergualin has yielded exceptional numbers (54%) of stable tolerant kidney allograft recipients, surviving over 6 years without rejection or immunosuppression. An early increase in IL-10 and reduction in IFNgamma distinguished recipients that subsequently became tolerant. Furthermore, analysis suggested that this immune switch was programmed within hours of transplantation. Administering deoxyspergualin within 5 h of surgery gave a higher incidence of tolerance (76%) compared to administration >5 h before or after surgery (11%, P<0.01). Deoxyspergualin inhibits nuclear translocation of activated NF-kappaB through heat shock proteins. Lymph node biopsies from tolerant recipients showed significant reductions in cytoplasmic expression of Hsp70 and RelB and almost complete inhibition of nuclear translocation of both. The early timing effect of deoxyspergualin suggests a crucial limitation to induction of stable tolerance is activation of Hsp-dependent innate responses to damage by ischemia-reperfusion. This was supported by studies in murine kidney reperfusion injury, where deoxyspergualin given 5 h before reperfusion protected renal function and reduced levels of IL-6 and IL-12. The narrow timing window for initiating deoxyspergualin treatment suggests the innate immune system is poised to defeat allograft tolerance induction, so effective blockade of NF-kappaB-mediated innate immunity must be in place early, to enable development of a tolerogenic environment.

Animals↗