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To investigate the effect of high-dose glucose-insulin-potassium (GIK) infusion on the outcomes of ST-elevation myocardial infarction (STEMI) in China.As part of the international multicentre CREATE-ECLA study, 7510 patients with STEMI, aged (62 +/- 12), presenting their symptoms within 12 hours of onset who were hospitalized in 274 centers throughout China from July 2001 through July 2004 were randomized to receive GIK intravenous infusion for 24 hours plus routine treatment (3739) or control group (n=3771) receiving routine treatment alone. The patients were flowed up in the out-patient department 30 days after the randomization to assess the rates of mortality, cardiac arrest, cardiogenic shock and re-infarction.The median time from symptom onset to randomization was 5.8 hours. The mortality of the control group was 10.4%, not significantly different from that of the GIK group (11.2%, hazard ratio = 1.05, 95% CI: 0.916-1.207, P = 0.476). There rates of cardiac arrest, cardiogenic shock, and re-infarction of the GIK group were 0.8%, 6.8%, and 2.0% respectively, all not significantly different from those of the control group (1.0%, 6.4%, and 1.9% respectively, all P0.05). At the Day 7 the heart failure rate of the GIK group was 19.7%, not significantly different from that of the control group (18.3%, P = 0.102). The symptomatic hypotension rate of the GIK group was 3.7%, significantly higher than that of the control group (1.2%, P0.01). The phlebitis rate of the GIK group was 2.2%, significantly higher than that f the control group (0.1%, P0.01). The net increased fluid volume of the control group wasl3 584 ml. more than that of the GIK group (1036 ml).High dose GIK infusion has neutral effect on mortality, cardiac arrest or cardiogenic shock in patients with acute STEMI in China.

To evaluation the effect of different mucosal vaccination pathway on hantavirus with the recombinant E. coli heat-labile enterotoxin B subunit (rLTB) as adjuvant.The rLTB was expressed and purified. Take the inactivated hantavirus strain 84Fli as vaccine, and immunized C57 BL/6 mice through intranasal, oral and vaginal respectively. Specific IgG and sectory IgA were detected by ELISA in serum, and vaginal washing samples respectively.The rLTB was efficiently expressed under the induction of lactose, identified by western blotting and GM-1 binding experiment. The vaccination through intranasal, oral and vaginal, can induce IgG and sectory IgA response.Inactivated hantavirus can produce mucosal immune response with rLTB as adjuvants through intranasal, oral and vaginal vaccination respectively.

To observe the ability of dengue virus recombinant envelope protein domain expressed in E. coli to inhibit virus infection and induce the neutralizing antibody.E III protein of Dengue virus serotypes 1-4 were expressed in E. coli BL21(DE3) then purified. Recombinant proteins were tested to inhibit DV2 from infecting BHK-21 cell. Rabbits were immunized with recombinant proteins to produce anti-E III serum. Antibody titers were determined by neutralizing assay.The recombinant E III proteins of Dengue virus serotypes 1-4 were expressed in E. coli. They effectively protected BHK cells in culture against DV2 infection. All four type anti-E III sera can neutralize DV2 but their efficacies are different.proteins of dengue virus expressed in E. coli can directly inhibit DV2 infection. Neutralizing antibodies were induced by E III proteins. Both E III protein of dengue virus and the neutralizing antibodies they induced are more efficient in inhibiting homologous dengue serotypes infection than heterologous serotypes.

Mucosal epithelial cells in the respiratory tract act as the first line of host innate defense against inhaled microbes by producing a range of molecules for clearance. In particular, epithelial mucins facilitate the mucociliary clearance by physically trapping the inhaled microbes. Up-regulation of mucin production thus represents an important host innate defense response against invading microbes. Excess mucin production, however, overwhelms the mucociliary clearance, resulting in defective mucosal defenses. Thus, tight regulation of mucin production is critical for maintaining an appropriate balance between beneficial and detrimental outcomes. Among various mechanisms, negative regulation plays an important role in tightly regulating mucin production. Here we show that the PAK4-JNK signaling pathway acted as a negative regulator for Streptococcus pneumoniae pneumolysin-induced MUC5AC mucin transcription. Moreover pneumolysin also selectively induced expression of MKP1 via a TLR4-dependent MyD88-TRAF6-ERK signaling pathway, which inhibited the PAK4-JNK signaling pathway, thereby leading to up-regulation of MUC5AC mucin production to maintain effective mucosal protection against S. pneumoniae infection. These studies provide novel insights into the molecular mechanisms underlying the tight regulation of mucin overproduction in the pathogenesis of airway infectious diseases and may lead to development of new therapeutic strategies.

Fucoidan, a marine sulfated polysaccharide has both antithrombotic and anti-inflammatory effects. We determined the effect of fucoidan on MUC5AC expression in a human bronchial epithelial cell line, NCI-H292. Reverse transcription-polymerase chain reaction (RT-PCR) analysis showed that fucoidan inhibited MUC5AC expression and protein secretion in cells stimulated with acrolein, a toxic aldehyde present in tobacco smoke. The activation of both nuclear factor-kappa B (NF-kappa B) and activator protein 1 (AP-1) are key steps in the transcriptional activation of MUC5AC. We found that the acrolein-mediated transactivation of MUC5AC was selectively dependent on AP-1 activation and was suppressed by fucoidan. Fucoidan-induced AP-1 inhibition and MUC5AC repression might be associated with fucoidan's protective effects against respiratory diseases.

The TGF-beta (transforming growth factor-beta) pathway represents an important signalling pathway involved in regulating diverse biological processes, including cell proliferation, differentiation and inflammation. Despite the critical role for TGF-beta in inflammatory responses, its role in regulating NF-kappaB (nuclear factor-kappaB)-dependent inflammatory responses still remains unknown. In the present study we show that TGF-beta1 synergizes with proinflammatory cytokine TNF-alpha (tumour necrosis factor-alpha) to induce NF-kappaB activation and the resultant inflammatory response in vitro and in vivo. TGF-beta1 synergistically enhances TNF-alpha-induced NF-kappaB DNA binding activity via induction of RelA acetylation. Moreover, synergistic enhancement of TNF-alpha-induced RelA acetylation and DNA-binding activity by TGF-beta1 is mediated by PKA (protein kinase A). Thus the present study reveals a novel role for TGF-beta in inflammatory responses and provides new insight into the regulation of NF-kappaB by TGF-beta signalling.

Toll-like receptors (TLRs) are important sensors of microbial pathogens and mediators of innate immune responses. Although the signal transduction of TLRs is well elucidated, their basal regulation is largely unexplored. Here we show that the tumor suppressor p53 positively regulates the transcription of TLR3, a receptor for viral double-stranded RNA and poly(I-C), by binding to the p53 site in the TLR3 promoter. TLR3 expression was lower in HCT116 p53(-/-) cells than in HCT116 p53(+/+) cells. Activation of p53 by 5-fluorouracil increased the TLR3 mRNA in epithelial cell lines with wild-type p53 but not in cell lines harboring mutant p53. Knockdown of p53 by small interfering RNA decreased the TLR3 expression. TLR3 mRNA was also lower in liver and intestine of p53(-/-) mice than in p53(+/+) mice. Furthermore, the poly(I-C)-induced phosphorylation of IkappaB-alpha, nuclear translocation of NF-kappaB, and phosphorylation of interferon regulatory transcription factor 3, were drastically reduced in HCT116 p53(-/-) cells, indicating a dysregulation of the two signaling pathways governed by TLR3. Consequently, induction of interleukin-8 and beta interferon after poly(I-C) stimulation was impaired in HCT116 p53(-/-) cells. These results suggest that p53 influences TLR3 expression and function and highlight a role of p53 in innate immune response in epithelial cells.

Bacteraemic pneumonia is a common cause of sepsis in critically ill patients today and is characterized by dysregulation of inflammation. The genetic factors predisposing to bacteraemic pneumonia are not yet fully understood. Innate immunity is pivotal for host defence against invading bacteria, and nuclear factor-kappa B (NF-kappaB) is central to bacteria-induced inflammation and immune responses. The deubiquitinating enzyme CYLD has been identified as a key negative regulator for NF-kappaB. In the present study, we investigated the role of CYLD in innate immune response in Escherichia coli pneumonia. Upon E. coli inoculation, Cyld(-/-) mice were hypersusceptible to E. coli pneumonia with higher mortality. Innate immune response to E. coli was enhanced in Cyld(-/-) cells and mice. Cyld(-/-) cells exhibited enhanced NF-kappaB activation upon E. coli inoculation, and the enhanced NF-kappaB activation by E. coli was abolished by perturbing IkappaB kinase (IKK) signalling. Furthermore, IKK inhibitor rescued Cyld(-/-) mice from lethal infection during E. coli pneumonia along with reduced inflammation. Taken together, these data showed that CYLD acts as a crucial negative regulator for E. coli pneumonia by negatively regulating NF-kappaB. These findings provide novel insight into the regulation of bacteraemic pneumonia and related diseases and may help develop novel therapeutic strategies for these diseases.

An investigation was made on the species composition and eco-type of Hemiptelea davidii community on Kerqin sandy land. The results showed that in study area, no shrub layer existed in H. davidii forest, while arbor layer could be divided into two sub-layers, with a height of 4.05-7.86 m for the upper layer, and of 2.05-3.20 m for the lower layer. In the community, there were 32 herbal species belonging to 27 genera of 13 families and dominant by Poaceae, Fabaceae and Compositae, and 11 areal types, among which, the species number in Mongolian-Northeastern China-Dahuricia-North China areal type was the highest (34.38%), followed by that in Northeastern China-North China areal type (12.5%). Among three water ecological types, mesophytes occupied 59.37% of the total, and mesoxerophytes and xerophytes occupied 25% and 15.63%, respectively. Hemicryptophytes had a larger amount in six life forms, accounting for 31.25% of the total. H. davidii community had the typical life form characteristics of temperate steppe.

The binding mechanism of human serum albumin with 3,3', 4', 7-tetradroxyflavone (FIS) was characterized by fluorescence and UV absorption spectra. The intrinsic fluorescence of HSA was significantly quenched by FIS in the physiological condition (pH 7.4), and the quenching mechanism is mainly static quenching process in the drug to protein molar ratio ranging from 0.1 to 10. The results revealed that the drug and protein formed 1 : 1 complex with the binding constants of (1.05 +/- 0.18) x 10(5) L x mol(-1). The dissociation behaviors of FIS in different pH conditions were investigated by UV absorption spectra, and it was found that FIS existed in the mixtures of ionic and neutral species. The UV absorption band I of FIS has a significant red shift (above 40 nm) after combination with HSA, demonstrating that FIS was bound to protein in ionic species which was driven by electrostatic force. The second derivative spectra of FIS showed that the combination included specific and non-specific forms. The intrinsic fluorescence of FIS was conspicuously enhanced in the presence of HSA due to the excited-state proton transfer (ESPT) and this further confirmed the complex formation of HSA with FIS.

To develop and improve a MacELISA method for the early diagnosis of hemorrhagic fever with renal syndrome (HFRS) with simplified operation procedure.The nucleic proteins of hantavirus were labeled with horse raddish peroxidase (HRP) and used as detection antigens. A two-step MacELISA based HRP conjugated antigen was established and the detection sensitivity and specificity were compared with commonly used three-step MacELISA.This method could be used to detect hantanvirus specific IgM with high sensitivity and specificity from human patient serum. There was not significant difference from commonly used three-step MacELISA and the sensitivity and specificity were 100%.This method is simple, sensitive and rapid in operation, and therefore could be used for the early diagnosis of HFRS.

To develop and evaluate a method for detection of the total antibodies against hemorrhagic fever with renal syndrome (HFRS) virus with improved sensitivity and simplified operation procedure.The nucleic proteins of hantavirus were used as coating antigens as well as detection antigens labeled with horse radish peroxidase (HRP). The operation protocol was established, optimized and compared with indirect fluorescence assay (IFA).The specificity of this method was 100 percent in the test of different human sera and 4-8 times more sensitive than IFA. And, it is simpler without requiring any change of the reagents, different sources of samples did not affect the results of the test.This method is specific, sensitive and simple for detection of the total antibodies in sera against hantavirus, could be used for the screening of Hantavirus infection in human and host rodent animals.

Shear stress–induced extracellular signal-regulated kinase (ERK)5 activation and the consequent regulation of Kruppel-like factor 2 and endothelial nitric oxide synthase expression represents one of the antiinflammatory and vascular tone regulatory mechanisms maintaining normal endothelial function. Endothelial dysfunction is a major initiator of atherosclerosis, a vascular pathology often associated with diabetes. Small ubiquitin-like modifier (SUMO) covalently attaches to certain residues of specific target transcription factors and could inhibit its activity. We investigated whether H 2 O 2 and AGE (advanced glycation end products), 2 well-known mediators of diabetes, negatively regulated ERK5 transcriptional activity and laminar flow–induced endothelial nitric oxide synthase expression through ERK5 SUMOylation. H 2 O 2 and AGE induced endogenous ERK5 SUMOylation. In addition, ERK5 SUMOylation was increased in the aortas from diabetic mice. ERK5 transcriptional activity, but not kinase activity, was inhibited by expression of Ubc9 (SUMO E2 conjugase) or PIAS1 (E3 ligase), suggesting the involvement of ERK5 SUMOylation on its transcriptional activity. Point-mutation analyses showed that ERK5 is covalently modified by SUMO at 2 conserved sites, Lys6 and Lys22, and that the SUMOylation defective mutant of ERK5, dominant negative form of Ubc9 (DN-Ubc9), and small interfering RNA PIAS1 reversed H 2 O 2 and AGE–mediated reduction of shear stress–mediated ERK5/myocyte enhancer factor 2 transcriptional activity, as well as promoter activity of Kruppel-like factor 2. Finally, PIAS1 knockdown reversed the inhibitory effect of H 2 O 2 in shear stress–induced Kruppel-like factor 2 and endothelial nitric oxide synthase expression. These data clearly defined SUMOylation-dependent ERK5 transcriptional repression independent of kinase activity and suggested this process as among the molecular mechanisms of diabetes-mediated endothelial dysfunction.

Mucin overproduction is a hallmark of chronic respiratory diseases (CRD) such as chronic obstructive pulmonary disease and asthma, and otitis media. Despite the fact that nontypeable Haemophilus influenzae (NTHi) and Streptococcus pneumoniae are co-existing under these disease conditions, little is known about how NTHi and S. pneumoniae induce mucin overproduction. Here we show that NTHi and S. pneumoniae, when present together, synergistically induce MUC5AC mucin transcription. TLR2/4-MyD88-TAK1 signaling cascade transmits signal to regulate the synergistic induction of MUC5AC. The activation of MKK3/6-p38 and ERK MAPK pathways are required for the synergistic induction of MUC5AC. Moreover, S. pneumoniae synergizes with NTHi to induce MUC5AC expression via AP-1-dependent mechanism. Thus, our studies provide direct evidence for the synergistic induction of MUC5AC in mixed infections and bring novel insights into our understanding of molecular mechanisms underlying polymicrobial infections in CRD and OM.

To develop a chemiluminescent enzyme-linked immunosorbent assay (CLEIA) for the detection of HTNV IgM antibody.Black solid 96 well microplate was coated with anti-human IgM-microantibody, HRP labeled HTNV recombinant nucleotide antigen was used as detection antigen, luminol-H2O2 was used as substrate, a CLEIA was established for the detection of HFRS patient serum IgM antibody and comparison of detection sensitivity, specificity, and stability were made between CLEIA and MacELISA.Correlate coefficient of CLEIA with MacELISA is 0.97; detection sensitivity of CLEIA is 100 percent while that of MacELISA is 92.1 percent; detection specificity of CLEIA and MacELISA are both 100 percent; coefficient of variance for intra-assay and inter-assay of CLEIA are both less than 15 percent, which are comparative with MacELISA.The established method of CLEIA is a sensitive, selective, and stable method; it is suitable for the early detection of HFRS patient serum IgM antibody.

The incidence of mixed viral/bacterial infections has increased recently because of the dramatic increase in antibiotic-resistant strains, the emergence of new pathogens, and the resurgence of old ones. Despite the relatively well-known role of viruses in enhancing bacterial infections, the impact of bacterial infections on viral infections remains unknown. In this study, we provide direct evidence that nontypeable Haemophilus influenzae (NTHi), a major respiratory bacterial pathogen, augments the host antiviral response by up-regulating epithelial Toll-like receptor 7 (TLR7) expression in vitro and in vivo. Moreover, NTHi induces TLR7 expression via a TLR2-MyD88-IRAK-TRAF6-IKK-NF-kappaB-dependent signaling pathway. Interestingly, CYLD, a novel deubiquitinase, acts as a negative regulator of TLR7 induction by NTHi. Our study thus provides new insights into a novel role for bacterial infection in enhancing host antiviral response and further identifies CYLD for the first time as a critical negative regulator of host antiviral response.

Mucin, a major component of mucus, plays a critical role in host mucosal defense response by participating in mucociliary clearance. However, if overproduced, overproduced mucus leads to airway mucus obstruction and conductive hearing loss. Despite extensive studies that focus on investigating how MUC5AC mucin is regulated by one inducer at a time, how MUC5AC is synergistically regulated by multiple factors remains unknown. Here we provide direct evidence for the first time that bacterial pathogen NTHi and human growth factor EGF synergize with each other to potently up-regulate MUC5AC mucin transcription. Moreover, activation of both p38 and ERK is required for synergistic induction of MUC5AC by NTHi and EGF. Finally, PAK2 and PAK4 are differentially involved in this synergistic induction of MUC5AC by acting upstream of p38 and ERK. Our studies bring novel insights into our understanding of synergistic regulation of MUC5AC mucin by both pathological and physiological inducers.

In this double-blinded, randomized, parallel study, we investigated the clinical efficacy of intravenous Acehytisine Hydrochloride (AHH) and propafenone on terminating paroxysmal supraventricular tachycardia (PSVT).Patients (18 - 70 years old) with either spontaneous or induced sustained supraventricular tachycardia lasted at least 15 min were recruited in this study. Exclusion criteria included sick sinus syndrome, atrial ventricular block or intraventricular block, etc. Eligible patients were randomly assigned to receive intravenously AHH (n=101) or propafenone (n=100) according to a proportion of 1:1 in a double-blinded manner. AHH (4 mg/kg, iv.) or propafenone (PRO, 1 mg/kg, iv.) was administered in 5 min followed by the same dose if no response was observed. Conversion times, vital signs, electrocardiograms were documented before and after drug administration.Except for age, the demographic characteristics and clinical features were comparable between the two groups. Efficacy on PSVT termination was comparable between AHH (72/101, 71.3%) and PRO group (73/100, 73.0%, P=0.6368). The average time from drug administration to conversion was also similar [AHH: (9.62 +/- 8.39) min vs. PRO: (10.61 +/- 9.47) min, P=0.5035]. In the AHH group, 59/72 episodes of PSVT were terminated by the first dose, and 66/72 were terminated prematurely. The average AHH dose in the 72 converted patients was (273.7 +/- 111.2) mg. In the PRO group, 54/73 episodes of PSVT were terminated by the first dose. The electrocardiographic parameters, such as sinus recovery time, longest PP and RR interval, PR interval, QRS interval, QT interval after conversion were similar between the two groups. Transient adverse events were reported in 11/101 (10.9%) patients in the AHH group and in 18/100 (18.0%,) in the PRO group (P=0.1653).With the dosage used in the present study, the efficacy on terminating PSVT was comparable between AHH and PRO.

Summary Streptococcus pneumoniae ( S. pneumoniae ) causes high early mortality in pneumococcal pneumonia, which is characterized by acute lung injury (ALI). The molecular mechanisms underlying ALI and the high early mortality remain unknown. Despite recent studies that identify deubiquitinating enzyme cylindromatosis (CYLD) as a key regulator for T cell development, tumor cell proliferation, and NF-κB transcription factor signaling, its role in regulating bacteria-induced lethality, however, is unknown. Here, we showed that CYLD deficiency protected mice from S. pneumoniae pneumolysin (PLY)-induced ALI and lethality. CYLD was highly induced by PLY, and it inhibited MKK3-p38 kinase-dependent expression of plasminogen activator inhibitor-1 (PAI-1) in lung, thereby potentiating ALI and mortality. Thus, CYLD is detrimental for host survival, thereby indicating a mechanism underlying the high early mortality of pneumococcal pneumonia.

To observe the electrophysiological effect of Acehytisine Hydrochloride (AHH) in Wu Zhi Shan (WZS) micropigs with experimental acute coronary occlusion.Adult WZS micropigs were randomized into group A: coronary ligation with AHH infusion (n = 9); group B: AHH infusion without coronary ligation (n = 9) and group C: coronary ligation with saline (NS) infusion (n = 9). Surface ECGs and cardiac electrophysiological data including atrium, atrium-ventricle junction and ventricle electrograms were collected by programmed electrical stimulation at ischemic baseline and after AHH (or NS) infusion.Compared to animals treated with AHH without ischemia, VARC-ERP was significantly increased while QT, QTc intervals, VRRP and VFRP were significantly reduced in ischemic animals treated with AHH. Compared to ischemic animals treated with saline, AHH prolongs the P-wave duration and PR interval, shortens QTc interval, prolongs ARP and AEP, also prolongs V-A reverse conduction time and VARC-ERP but shortens VFRP. No proarrhythmia effect was found in both AHH treated groups.AHH resulted in significant electrophysiological effects on this porcine acute coronary ischemic model.

Transforming growth factor-beta (TGF-beta) family members are multifunctional growth factors involved in regulating diverse biological processes. Despite the critical role for TGF-beta in regulating cell proliferation, differentiation, migration and development, its role in regulating NF-kappaB-dependent inflammatory response still remains unclear. Here, we show that TGF-beta1 induces acetylation of NF-kappaB p65 subunit to synergistically enhance bacterium nontypeable Haemophilus influenzae-induced NF-kappaB activation and inflammatory response in vitro and in vivo. The TGF-beta1-induced acetylation of p65 is mediated via a Smad3/4-PKA-p300-dependent signaling pathway. Acetylation of p65 at lysine 221 by TGF-beta1 is critical for synergistic enhancement of bacteria-induced DNA-binding activity, NF-kappaB activation, NF-kappaB-dependent transcription of TNF-alpha and IL-1beta and interstitial polymorphonuclear neutrophil infiltration in vitro and in vivo. These studies provide new insights into the novel regulation of NF-kappaB by TGF-beta signaling.

Airway diseases such as asthma and chronic obstructive pulmonary disease (COPD) are characterized by excessive inflammation and are exacerbated by nontypeable Haemophilus influenzae (NTHi). Airway epithelial cells mount the initial innate immune responses to invading pathogens and thus modulate inflammation. While inflammation is necessary to eliminate a pathogen, excessive inflammation can cause damage to the host tissue. Therefore, the inflammatory response must be tightly regulated and deciphering the signaling pathways involved in this response will enhance our understanding of the regulation of the host inflammatory response. NTHi binds to TLR2 and signal propagation requires the adaptor molecule myeloid differentiation factor 88 (MyD88). An alternative spliced form of MyD88 is called MyD88 short (MyD88s) and has been identified in macrophages and embryonic cell lines as a negative regulator of inflammation. However, the role of MyD88s in NTHi-induced inflammation in airway epithelial cells remains unknown. Here we show that NTHi induces MyD88s expression and MyD88s is a negative regulator of inflammation in airway epithelial cells. We further demonstrate that MyD88s is positively regulated by IKKβ and CREB and negatively regulated by ERK1/2 signaling pathways. Taken together these data indicate that airway inflammation is controlled in a negative feedback manner involving MyD88s and suggest that airway epithelial cells are essential to maintain immune homeostasis.

To analyze the current use of reperfusion strategies and the outcomes of patients with ST elevation acute coronary syndromes (ACS) in China.A total of 518 consecutive patients (371 male and 147 females, mean age 65 +/- 11) with ST elevation ACS or newly discovered left bundle branch block were registered from 20 hospitals from 5 regions (ranging from large regional centre hospitals to small county hospitals) in China. Patient general characteristics, reperfusion patterns and outcomes were analyzed. Patients were followed up for 3 months.The median time from pain onset to presentation at the hospital was 4 hours. Pre-hospital delay12 hours was found in 20% patients. Fifty-six percent patients (292/518) underwent reperfusion therapy (134 with primary percutaneous coronary intervention and 158 with fibrinolysis). The median time from admission to reperfusion (door-to-needle) was 65 min in fibrinolysis group and 110 min (door-to-cath) in primary PCI group respectively. Urokinase was used in 67% (106/158) patients underwent fibrinolysis. Multivariate logistic regression analysis showed that age/= 75 years (P0.01), previous myocardial infarction (P0.01) and history of congestive heart failure (P0.05) were associated with no reperfusion therapy. Mortality and congestive heart failure rates were significantly higher in patients with no reperfusion therapy not only at discharge (P0.01) but also at 3 months (P0.01) compared to patients underwent reperfusion. The incidence of combined outcomes (death or MI, and death, MI or Strobe) was also higher in patients without reperfusion therapy at 3 months (all P0.01) compared to patients underwent reperfusion. There were no differences on combined outcomes between fibrinolysis and primary PCI subgroups.Reperfusion therapy was the primary treatment of choice to improve the outcomes of patients with ST elevation ACS. Strategies to increase reperfusion therapy rate for ST elevation ACS are urgently needed in China.

In review of the past studies on NF-kappaB regulation, most of them have focused on investigating how NF-kappaB is activated by a single inducer at a time. Given the fact that, in mixed bacterial infections in vivo, multiple inflammation inducers, including both nontypeable Haemophilus influenzae (NTHi) and Streptococcus pneumoniae, are present simultaneously, a key issue that has yet to be addressed is whether NTHi and S. pneumoniae simultaneously activate NF-kappaB and the subsequent inflammatory response in a synergistic manner. Here, we show that NTHi and S. pneumoniae synergistically induce NF-kappaB-dependent inflammatory response via activation of multiple signaling pathways in vitro and in vivo. The classical IKKbeta-IkappaBalpha and p38 MAPK pathways are involved in synergistic activation of NF-kappaB via two distinct mechanisms, p65 nuclear translocation-dependent and -independent mechanisms. Moreover, casein kinase 2 (CK2) is involved in synergistic induction of NF-kappaB via a mechanism dependent on phosphorylation of p65 at both Ser536 and Ser276 sites. These studies bring new insights into the molecular mechanisms underlying the NF-kappaB-dependent inflammatory response in polymicrobial infections and may lead to development of novel therapeutic strategies for modulating inflammation in mixed infections for patients with otitis media and chronic obstructive pulmonary diseases.

To investigate the epidemiological features of human calicivirus( HuCV) infection in children with diarrhea in a hospital in Guangzhou.Stool specimens were collected from children with viral diarrhea diagnosed between October, 2003 and January, 2004 and between October, 2004 and January, 2005. HuCV was detected by means of RT-PCR and sequence analysis of the PCR products.Eighty specimens positive for Norwalk-like viruses (NLVs) were identified from 648 stool specimens, with a positivity rate of 12.35%, and sapporo-like viruses (SLVs) were identified in 2 specimens (0.31%). The monthly NLV positivity rate was 11.74% (Oct.), 14.16% (Nov.), 9. 09% (Dec.) and 13.95% (Jan.), respectively, showing no significant variation in these months. NLVs mainly infected children below 2 years old. Twenty-two strains of NLVs were sequenced and analyzed, and 21 of them were identified as GII and the genotype of 1 strain could not be determined. The prevalent viral population were GII-3 and GII-4 in 2003 and was GII-4 in 2004, and both of the SLVs belong to GI-1.NLVs is one of the important pathogens causing sporadic acute gastroenteritis in children admitted in the hospital in Guangzhou, and the prevalent strains are GII-3 and GII-4 , but different prevalent strains are possible in different periods.

The transforming growth factor beta (TGF-beta) pathway represents an important signaling pathway involved in the regulation of diverse biological processes, including cell proliferation, differentiation, and apoptosis. Despite the known role of TGF-betaR-mediated signaling in suppressing immune response, its role in regulating human Toll-like receptors (TLRs), key host defense receptors that recognize invading bacterial pathogens, however, remains unknown. Here, we show for the first time that TGF-betaR-Smad3/4 signaling pathway acts as a positive regulator for TLR2 induction by bacterium nontypeable Hemophilus influenzae (NTHi) in vitro and in vivo. The positive regulation of TLR2 induction by TGF-betaR is mediated via a dual mechanism involving distinct signaling pathways. One mechanism involves functional cooperation between the TGF-betaR-Smad3/4 pathway and NF-kappaB pathway. Another involves MAP kinase phosphatase 1 (MKP-1)-dependent inhibition of p38 MAPK, a known negative regulator for TLR2 induction. Moreover, we showed that TbetaR-mediated signaling is probably activated by NTHi-derived TGF-beta mimicry molecule via an autocrine-independent mechanism. Thus, our study provides new insights into the role of TGF-beta signaling in positively regulating host defense response by tightly controlling the expression level of TLR2 during bacterial infections and may lead to new therapeutic strategies for modulating host defense and inflammatory response.

To explore the epidemiological characteristics of Norwalk-like virus (NLVs) infection in children with diarrhea and to study the genotype and predominant cluster at a hospital in Guangzhou city.Fecal specimens from 358 children with acute gastroenteritis from October 2003 to January 2004 and information about the cases were collected. NLVs was detected from the specimens by reverse transcription-polymerase chain reaction (RT-PCR) and the PCR products were purified and sequenced.Forty-two positive specimens were detected from the 358 fecal specimen with a positive rate of 11.73% (42/358). Of these, 40 specimens were obtained from infants younger than 3 years of age. The youngest infant infected with NLVs in this study was only 25 days. The positive rate in November (17.27%) was the highest. Eleven positive PCR products were selected and sequenced. Nucleotide sequence analysis revealed that 11 strains all belong to genogroup II (G II), and of these, 5 strains belonged to G II-3 cluster, with another 5 strains belonged to G II-4 cluster. However, one strain with its cluster could not be determined.NLVs served as one of the important pathogens causing sporadic acute gastroenteritis among children at a hospital in Guangzhou. The predominant strains were identified as G II-3 and G II-4 cluster.

To analyze the clinical characteristics and management of the patients with ST segment elevation myocardial infarction (STEMI) in China.As part of the international multicentre CREATE Study, the clinical data of 7510 patients with STEMI presenting their symptoms within 12 hours of onset who were hospitalized in 274 centers throughout China from July 2001 through July 2004, aged 62.7, were collected to be analyzed.99.3% of these patients had STEMI, and 0.7% had new left bundle-branch block. 11.5% of them underwent percutaneous coronary intervention (PCI), 52.5% underwent thrombolytic therapy, and 0.1% coronary artery bypass grafting. All types of reperfusion therapy were carried out to 62.4% of the patients during the hospitalization. The medication therapy used was similar to those in the previous Registry report. The composite of all-cause mortality, re-infarction, and stroke within the first 7 days was 10.3% and the all-cause mortality within 30 days was 11.1%.The clinical characteristics and management of the STEMI patients in China have been described. The age of onset of STEMI is 62 years on average. The most common complication is hypertension, and diabetes mellitus is relatively rare. rt-PA was used rarely mainly because of the expensive price. A large proportion of patients in China receive reperfusion, which is worth recommendation. Active intervention should be carried out in the early stage after infarction.

To evaluate the safety of recombinant human interferon alpha-2b for nasal spray for the prevention of SARS and other upper respiratory viral infections.Field epidemiologic evaluation was conducted, the design was randomized and had a synchronously parallel control group. In the study, the drugs were given for five days and all subjects were followed up for ten days.During the period of using interferon, body temperature of the experimental group was normal compared to the control group. Experimental group had more influenza-like symptoms than the control group (P0.05), such as headache (4.83%-7.09%), dizziness (7.17%-11.63%), lassitude (8.55%-15.06%), muscular soreness (4.43%-7.09%), pharynx dryness (12.10%-17.85%), angina (6.25%-8.72%), abdominal pain (2.30%-5.50%) and diarrhea (2.45%-5.66%). Most of side effects reached their peak with in the first 3 days. Except for pharynx dryness, the incidences of all other side effects declined after completion of the use of the trial drug, and incidences of some symptoms in experimental group were lower than those of the control group. There were no significant differences in the symptoms of cough and expectoration between the experimental group and the control group. The incidence of exanthem in the control group was significantly higher than that in the experimental group. The side effect of bloody nasal mucus was not observed in experimental group, which had been reported by other authors in several volunteer studies.Using recombinant human interferon alpha-2b for nasal spray could lead to some influenza-like symptoms, however, all those symptoms were mild , reversible, and relieved after completion of the use of the trial drug. No serious side effects were found during the period of following up. The authors conclude that the drug is safe.

To study the protective effects of Panax notoginseng saponins (PNS) on angiotensin II (Ang II)-induced rat cardiomyocyte apoptosis in vitro and the probable mechanism.Cultured cardiomyocytes from neonatal rats were stimulated with Ang II. Cell viability was measured by MTT. Apoptosis was evaluated using Acridine Orange (AO) fluorescent dye staining and flow cytometry; Fluo-3 AM was used to test the change of intracellular free calcium.It was found that incubating with Ang II (10(-7) mol x L(-1)) for 48 h increased cardiomyocyte apoptosis, PNS (25, 100 mg x mL(-1)) increased myocyte viability. PNS (50 mg x mL(-1)) significantly decreased this Ang II-induced rat cardiomyocyte apoptosis (P0.05) and decreased fluorescent intensity of intracellular calcium.PNS has a significant effect on Ang II-induced rat cardiomyocytes apoptosis in vitro by alleviating intracellular calcium overload.

To identify the influencing factors of surviving time of patients with non-ST elevation acute coronary syndromes in China.The data of patients with non-ST elevation acute coronary syndromes hospitalized in 38 hospitals in China, including clinical characteristics, therapeutic procedure, and major events at hospitalization and two years' follow-up period, were collected and analyzed as part of an international multicentre registry--OASIS. No particular intervention was needed in patients' therapy. The needed data were recorded by filling in the Case Report Forms offered by Canadian Cardiovascular Collaboration according to the protocol. Cox regression model was used to analyze the association with survival and multiple factors recorded.From April 1999 to December 2001, 2294 patients with non-ST elevation acute coronary syndromes were enrolled in 38 Chinese hospitals nationwide, two years' follow-up had been finished among 2188 of which with a mean age of 62.8 +/- 8.3 and 62.3% being male. The clinical diagnosis at admission was unstable angina in 88.5% of the patients and was non Q-wave myocardial infarction in the remaining 11.5%. The number of death totaled up to 174 with a mortality of 7.6% by the end of the two years' follow-up. The most common cause of death was severe arrhythmia or sudden death (92 cases, 52.9%). More than 70 factors had been analyzed by Cox regression model in order to determine which might influence survival. Risk factors that reduced lifetime were: frequencies of myocardial infarction during follow-up period, stroke during hospitalization, thrombolysis during hospitalization, frequencies of heart failure during follow-up period, heart failure during hospitalization, frequencies of stroke during follow-up period, history of myocardial infarction, times of hospitalization because of non-cardiovascular disease during follow-up period, history of diabetes mellitus, duration of the first hospitalization, and age by the first hospitalization. Protective factors that prolonged lifetime were the frequencies of using oral anticoagulant, anti-platelet medicine, nitrate, and lipid lowering agents during follow-up period.The most common cause of death in patients with non-ST elevation acute coronary syndromes is severe arrhythmias or sudden death in China. Several factors influence the survival of patients. The effects of most factors in this study are similar to those founded by previous evidence based medical study.

To carry out bioinformatic analysis of Vibrio parahaemolyticus thermolabile hemolysin gene (tlh) obtained by PCR amplification.The tlh gene amplified by PCR was cloned into the vector pET32a(+) and sequenced, followed by analysis of the biological information by with presenting the sequences to the websites of bioinformatics on the Internet.The sequenced tlh gene (named tlh14-90) was entered into GenBank with the accession number of AY289609. Tlh14-90 has a length of 1 257 bp with both start and stop codons, having 99% homology with the tlh gene of WP1. Tlh14-90 is predicted to encode a protein containing 418 amino acids (named TLH14-90, with the molecular formula of C(2131)H(3184)N(548)O(649)S(16), molecular weight of 47 392.9, and the theoretical PI of 4.92). This protein consists of 4 Cys and the contents of Ala, Leu and Asn are 11.0%, 7.4% and 7.2%, respectively, having a hydrophobic parameter of -34 calculated using kappa-D method. Predicted as a alpha/protein for its secondary structure, TLH14-90 has not been identified for its tertiary structure.

To identify the molecular biological characteristics of human caliciviruses (HuCVs) and new strains of norovirus (NV) detected in Guangdong Province.HuCVs were detected from the stool specimens by reverse transcription-PCR (RT-PCR), and the PCR products were cloned, transformed, and blasted with Genbank. The nucleotide sequence homology was also evaluated with PHYLIP package, and the phylogenetic tree was generated by Treeview.HuCVs were detected from 18 out of 206 stool specimens from children with viral diarrhea (8.74%), which were all confirmed subsequently as NVs. Two strains HuCV/NVGII 003/2003/CHN(32282) and HuCV/NVGII 004/2003/CHN(32283) from the selected 6 NVs strain were identified as new NV strains.New NV strains in consistency with the blood-type distribution of Chinese are present in Guangzhou, which have not been reported in other countries.

The transcription factor NF-kappaB regulates genes involved in inflammatory and immune responses, tumorigenesis, and apoptosis. In contrast to the pleiotropic stimuli that lead to its positive regulation, the known signaling mechanisms that underlie the negative regulation of NF-kappaB are very few. Recent studies have identified the tumor suppressor CYLD, loss of which causes a benign human syndrome called cylindromatosis, as a key negative regulator for NF-kappaB signaling by deubiquitinating tumor necrosis factor (TNF) receptor-associated factor (TRAF) 2, TRAF6, and NEMO (NF-kappaB essential modulator, also known as IkappaB kinase gamma). However, how CYLD is regulated remains unknown. The present study revealed a novel autoregulatory feedback pathway through which activation of NF-kappaB by TNF-alpha and bacterium nontypeable Haemophilus influenzae (NTHi) induces CYLD that in turn leads to the negative regulation of NF-kappaB signaling. In addition, TRAF2 and TRAF6 appear to be differentially involved in NF-kappaB-dependent induction of CYLD by TNF-alpha and NTHi. These findings provide novel insights into the autoregulation of NF-kappaB activation.

Background Despite its direct connection to the nasopharynx which harbors otitis media pathogens as part of its normal flora, the middle ear cavity is kept free of these bacteria by as yet unknown mechanisms. Respiratory mucosal epithelia, including those of the middle ear and eustachian tube, secrete antimicrobial effectors including lysozyme, lactoferrin and β defensins-1 and -2. To elucidate the role of these innate immune molecules in the normal defense and maintenance of sterility of respiratory mucosa such as that of the middle ear, we assessed their effect on the respiratory pathogens nontypeable Haemophilus influenzae (NTHi) 12, Moraxella catarrhalis 035E, and Streptococcus pneumoniae 3, and 6B. Methods Two assay methods, the radial assay and the liquid broth assay, were employed for testing the antimicrobial activity of the molecules. This was done in order to minimize the possibility that the observed effects were artifacts of any single assay system employed. Also, transmission electron microscopy (TEM) was employed to evaluate the effect of antimicrobial innate immune molecules on OM pathogens. For the statistical analysis of the data, Student's t-test was performed. Results Results of the radial diffusion assay showed that β defensin-2 was active against all four OM pathogens tested, while treatment with β defensin-1 appeared to only affect M. catarrhalis. The radial assay results also showed that lysozyme was quite effective against S. pneumoniae 3 and 6B and was partially bacteriostatic/bactericidal against M. catarrhalis. Lysozyme however, appeared not to affect the growth of NTHi. Thus, lysozyme seems to have a more pronounced impact on the growth of the Gram-positive S. pneumoniae as compared to that of Gram-negative pathogens. Lactoferrin on the other hand, enhanced the growth of the bacteria tested. The results of the radial assays were confirmed using liquid broth assays for antimicrobial activity, and showed that lysozyme and β defensin-2 could act synergistically against S. pneumoniae 6B. Moreover, in the liquid broth assay, β defensin-1 showed a modest inhibitory effect on the growth of S. pneumoniae 6B. As assessed by ultrastructural analysis, lysozyme and β defensin-2, and to a much lesser extent, β defensin-1, appeared to be able to cause damage to the bacterial membranes. Conclusions Here we report that lysozyme and the β defensins can inhibit the growth of clinical isolates of otitis media pathogens – namely NTHi strain 12, S. pneumoniae strains 3 and 6B and M. catarrhalis strain 035E – and cause ultrastructural damage to these pathogens. Moreover, we demonstrate that lysozyme and β defensin-2 can act synergistically against S. pneumoniae. These findings are consistent with the concept that secreted antimicrobial peptides and other components of innate immunity constitute the first line of defense protecting host mucosal surfaces, including the tubotympanal (eustachian tube and middle ear cavity) mucosa, against pathogens.

Myeloid elf-1-like factor (MEF) or Elf4, which is a member of the ETS transcription factor family, up-regulates the basal expression of lysozyme gene in epithelial cells and is constitutively localized in the nucleus. The mammalian cell nucleus is organized into distinct nuclear domains or compartments that are essential for diverse physiological processes. Promyelocytic leukemia (PML) nuclear body or nuclear domain 10 is one of the nuclear domains and is involved in tumor suppression and regulation of transcription. Here, we investigate the role of PML nuclear body in MEF transactivation. We show that PML, but not Sp100, induced the accumulation of MEF in PML nuclear bodies and that MEF and PML physically interacted. This interaction stimulated MEF transcriptional activity, resulting in the up-regulation of endogenous lysozyme expression. Amino acids 348-517 of MEF were required for the accumulation of MEF in PML nuclear bodies and up-regulation of lysozyme transcription, which is enhanced by PML. Moreover, the C-terminal region of MEF spanning amino acids 477-517 was the putative region required for interaction between MEF and PML as determined with the use of the mammalian two-hybrid system. In addition, heat-shock treatment induced the accumulation of MEF in endogenous PML nuclear bodies and enhanced MEF transactivation of lysozyme gene. Thus, the recruitment of MEF to PML nuclear bodies may partly regulate lysozyme transcription in epithelial cells.

To investigate the change of intestinal flora in subjects receiving oral vaccination with inactivated whole-cell/recombinant B subunit (WC/rBS) O139 cholera vaccine made in China.The fecal smears of 5 groups of subjects receiving the vaccine were collected before and 2 and 3 months after the vaccination to examine the distribution of the intestinal flora.In the 458 specimens examined, the total amount of bacteria and percentages of G+b, G+c and G-c were decreased whereas the amount of G-b significantly increased after vaccination. Compared with the placebo control group, the vaccinated groups exhibited significant changes in G+b and G-c, which also varied significantly between the group receiving 3 doses of WC/rBS vaccine and the other vaccinated groups. The vaccination produced distinct changes in the percentage of the intestinal flora at different time points, showing the most obvious effect on G-b. Oral vaccination was shown to affect the percentage of the intestinal flora very likely through its effect on the amount of G-b. The percentages of G+b, G-b and G-c varied significantly between male and female subjects after vaccination.Oral vaccination with WC/rBS affects the intestinal flora in human, and the mechanism as well as the implications needs to be further explored.

Background Despite the importance of glucocorticoids in suppressing immune and inflammatory responses, their role in enhancing host immune and defense response against invading bacteria is poorly understood. Toll-like receptor 2 (TLR2) has recently gained importance as one of the major host defense receptors. The increased expression of TLR2 in response to bacteria-induced cytokines has been thought to be crucial for the accelerated immune response and resensitization of epithelial cells to invading pathogens. Results We show that IL-1β, a key proinflammatory cytokine, greatly up-regulates TLR2 expression in human epithelial cells via a positive IKKβ-IκBα-dependent NF-κB pathway and negative MEKK1-MKK4/7-JNK1/2 and MKK3/6-p38 α/β pathways. Glucocorticoids synergistically enhance IL-1β-induced TLR2 expression via specific up-regulation of the MAP kinase phosphatase-1 that, in turn, leads to dephosphorylation and inactivation of both MAPK JNK and p38, the negative regulators for TLR2 induction. Conclusion These results indicate that glucocorticoids not only suppress immune and inflammatory response, but also enhance the expression of the host defense receptor, TLR2. Thus, our studies may bring new insights into the novel role of glucocorticoids in orchestrating and optimizing host immune and defense responses during bacterial infections and enhance our understanding of the signaling mechanisms underlying the glucocorticoid-mediated attenuation of MAPK.

NontypeableHaemophilus influenzae(NTHI) is an important etiological agent of otitis media (OM) and of exacerbated chronic obstructive pulmonary diseases (COPD). Inflammation is a hallmark of both diseases. Interleukin-8 (IL-8), one of the important inflammatory mediators, is induced by NTHI and may play a significant role in the pathogenesis of these diseases. Our studies demonstrated that a soluble cytoplasmic fraction (SCF) from NTHI induced much greater IL-8 expression by human epithelial cells than did NTHI lipooligosaccharides and envelope proteins. The IL-8-inducing activity was associated with molecules of ≤3 kDa from SCF and was peptidase and lipase sensitive, suggesting that small lipopeptides are responsible for the strong IL-8 induction. Moreover, multiple intracellular signaling pathways were activated in response to cytoplasmic molecules. The results indicated that the p38 mitogen-activated protein kinase (MAPK) and Src-dependent Raf-1-Mek1/2-extracellular signal-regulated kinase mitogen-activated protein kinase (ERK MAPK) pathways are required for NTHI-induced IL-8 production. In contrast, the phosphatidylinositol 3-kinase (PI3K)-Akt pathway did not affect IL-8 expression, although this pathway was concomitantly activated upon exposure to NTHI SCF. The PI3K-Akt pathway was also directly activated by IL-8 and significantly inhibited by an antagonist of IL-8 receptors during NTHI stimulation. These results indicated that the PI3K-Akt pathway is activated in response to IL-8 that is induced by NTHI and may lead to other important epithelial cell responses. This work provides insight into essential molecular and cellular events that may impact on the pathogenesis of OM and COPD and identifies rational targets for anti-inflammatory intervention.

To understand the molecular mechanisms of hantavirus assembly and maturation by stably expressing the intracellular antibodies to hantavirus glycoprotein G1 and G2 in endoplasmic reticulum (ER) and cytoplasm (Cyto) of Vero E6 cell.The genes of VH and VL of antibodies against glycoprotein of hantavirus were amplified by PCR and cloned into pOPE 101-215 (Yol) vector. The G1 and G2 proteins specific ScFv genes were first expressed in E.coli and the function and binding properties were identified. The gene of ScFv were further inserted into intracellular expression vectyrs pEF/ myc/ ER and pEF/ myc/ CYTO vector and transfected Vero E6 cell. The clonal cell line which stabl expresses ScFv were isolated under the pressure of G418.The ScFv genes of hantavirus G1 and G2 specific antibodies were successfully expressed in subcellular compartment ER and Cyto of Vero E6 cells and specifically targeted G1 and G2 protein after virus infection of the cells.The recombination of intrabody to Hantann virus glycoprotein was constructed successfully, and it may provide basic material for the studying antiviral gene therapy and the molecular mechanism of viral replication and infection.

In this experiment, it was designed to carry out proliferous culture of bovine blastocysts(day 7) derived from embryos cloned through bovine somatic cell nuclear transfer, isolating and passaging of ES cells. The cells of blastocysts, which were planted on feeder layer, formed small colonies within 24 h. The nest-shape colonies occurred after culturing for 2-3 days. After the colonies in the same shape were isolated and passaged 4-5 times, many different size colonies with monolayer of multi-cells appeared. The colonies that had been passaged 4-5 times were planted into 4-wells multi-dishes without feeder layer. The colonies with monolayer of multi-cells appeared after 24 h, spread all over the bottom of the dishes, emerged epidermis-like cells that appeared reticulate after 4-7 days. These cells were used as donor cells to carry out nuclear transfer. The results showed that 80% (40/50) of the reconstructed embryos cleaved, 5% (2/40) and 2.5% (1/40) of them developed to the morulaes and blastocyst stage, respectively. It revealed that ES-like cells derived embryos constructed through somatic cell nuclear transfer have the developmental potentials.

Background Recent studies demonstrate that there are sex differences in the expression of angiotensin receptor type 2 (AT2-R) in the kidney and that AT2-R plays an enhanced role in regulating blood pressure (BP) in females. Also, brain AT2-R activation has been reported to negatively modulate BP and sympathetic outflow. The present study investigated whether the central blockade of endogenous AT2-R augments deoxycorticosterone acetate (DOCA)/salt-induced hypertension in both male and female rats. Methods All rats were subcutaneously infused with DOCA combined with 1% NaCl solution as the sole drinking fluid. BP and heart rate (HR) were recorded by telemetric transmitters. To determine the effect of central AT2-R on DOCA/salt-induced hypertension, male and female rats were intracerebroventricularly (icv) infused with AT2-R antagonist, PD123,319, during DOCA/salt treatment. Subsequently, the paraventricular nucleus (PVN) of the hypothalamus, a key cardiovascular regulatory region of the brain, was analyzed by quantitative real-time PCR and Western blot. Results DOCA/salt treatment elicited a greater increase in BP in male rats than that in females. Icv infusions of the AT2-R antagonist significantly augmented DOCA/salt pressor effects in females. However, this same treatment had no enhanced effect on DOCA/salt-induced increase in the BP in males. Real-time PCR and Western blot analysis of the female brain revealed that DOCA/salt treatment enhanced the mRNA and protein expression for both antihypertensive components including AT2-R, angiotensin-converting enzyme (ACE)-2, and interleukin (IL)-10 and hypertensive components including angiotensin receptor type 1 (AT1-R), ACE-1, tumor necrosis factor (TNF)-α, and IL-1β, but decreased mRNA expression of renin in the PVN. The central blockade of AT2-R reversed the changes in mRNA and protein expressions of ACE-2, IL-10, and renin, further increased the expressions of TNF-α and IL-1β, and kept higher the expressions of AT1-R, ACE-1, and AT2-R. Conclusions These results indicate that endogenous AT2-R activation in the brain plays an important protective role in the development of DOCA/salt-induced hypertension in females, but not in males. The protective effect of AT2-R in females involves regulating the expression of brain renin-angiotensin system components and proinflammatory cytokines.

The recognition of invading microbes followed by the induction of effective innate immune response is crucial for host survival. Human surface epithelial cells are situated at host-environment boundaries and thus act as the first line of host defense against invading microbes. They recognize the microbial ligands via Toll-like receptors (TLRs) expressed on the surface of epithelial cells. TLR2 has gained importance as a major receptor for a variety of microbial ligands. In contrast to its high expression in lymphoid tissues, TLR2 is expressed at low level in epithelial cells. Thus, it remains unclear whether the low amount of TLR2 expressed in epithelial cells is sufficient for mediating bacteria-induced host defense and immune response and whether TLR2 expression can be up-regulated by bacteria during infection. Here, we show that TLR2, although expressed at very low level in unstimulated human epithelial cells, is greatly up-regulated by nontypeable Hemophilus influenzae (NTHi), an important human bacterial pathogen causing otitis media and chronic obstructive pulmonary diseases. Activation of an IKKbeta-IkappaBalpha-dependent NF-kappaB pathway is required for TLR2 induction, whereas inhibition of the MKK3/6-p38alpha/beta pathway leads to enhancement of NTHi-induced TLR2 up-regulation. Surprisingly, glucocorticoids, well known potent anti-inflammatory agents, synergistically enhance NTHi-induced TLR2 up-regulation likely via a negative cross-talk with the p38 MAP kinase pathway. These studies may bring new insights into the role of bacteria and glucocorticoids in regulating host defense and immune response and lead to novel therapeutic strategies for modulating innate immune and inflammatory responses for otitis media and chronic obstructive pulmonary diseases.

EMMPRIN (extracellular matrix metalloproteinase inducer) stimulates fibroblast metalloproteinases (MMP) 1, 2 and 3 (Kataoka et al. (1993) Cancer Res. 53, 3154–3158). Here we focus on MMP-1, showing that in lung tumors, MMP-1's cognate mRNA is strongly expressed in stromal fibroblasts adjacent to EMMPRIN-expressing tumor cells. In vitro, EMMPRIN upregulates MMP-1 mRNA expression in a concentration-dependent manner, with a peak accumulation at 24 h. The response is genistein-sensitive, suggesting it is dependent on tyrosine kinase activity. Analysis of tyrosine phosphorylation-dependent MAP kinases ERK 1/2, SAPK/JNK, and p38 showed that the activity of p38 but not that of the other 2 kinases was elevated in response to EMMPRIN. That p38 activity was required for EMMPRIN stimulation of MMP-1 was evident from results showing that the p38 inhibitor SB203580 blocked this response. This is the first available information regarding the mechanism by which tumor-associated molecules upregulate MMP synthesis in stromal fibroblasts.