Auricularia auricular-judae polysaccharide attenuates lipopolysaccharide-induced acute lung injury by inhibiting oxidative stress and inflammation.

Auricularia auricular-judae polysaccharide (AAP) has shown a variety of pharmacological properties. In the present study, the role of AAP in acute lung injury (ALI) induced by lipopolysaccharide (LPS) was analyzed in rats to further explore the possible underlying mechanisms. Adult Sprague-Dawley rats were randomly assigned into the control, AAP, LPS and LPS plus AAP groups. Rats were injected with LPS (10 mg/kg, intraperitoneal) to induce ALI. Rats in the LPS plus AAP group were treated with AAP for 7 days before the induction of ALI. The protein concentration in the bronchoalveolar lavage fluid (BALF) was measured. The animal lung edema degree was evaluated by the wet/dry (W/D) weight ratio. The myeloperoxidase (MPO) activity and malondialdehyde (MDA) level were assayed by MPO and MDA kits, respectively. The levels of inflammatory mediators, tumor necrosis factor-α (TNF-α) and interleukin (IL)-6, were assayed by the enzyme-linked immunosorbent assay method. Pathological changes of lung tissues were observed by hematoxylin and eosin staining. The data showed that treatment with AAP significantly improved LPS-induced lung pathological changes, attenuated protein concentration in the BALF, inhibited MPO activity and reduced the MDA level and lung W/D weight ratio. AAP also inhibited the release of TNF-α and IL-6 in blood. The results indicated that AAP has a protective effect on LPS-induced ALI in rats.

Trametinib, a novel MEK kinase inhibitor, suppresses lipopolysaccharide-induced tumor necrosis factor (TNF)-α production and endotoxin shock.

Lipopolysaccharide (LPS), one of the most prominent pathogen-associated molecular patterns (PAMPs), activates macrophages, causing release of toxic cytokines (i.e. tumor necrosis factor (TNF)-α) that may provoke inflammation and endotoxin shock. Here, we tested the potential role of trametinib, a novel and highly potent MAPK/ERK kinase (MEK) inhibitor, against LPS-induced TNF-α response in monocytes, and analyzed the underlying mechanisms. We showed that trametinib, at nM concentrations, dramatically inhibited LPS-induced TNF-α mRNA expression and protein secretion in transformed (RAW 264.7 cells) and primary murine macrophages. In ex-vivo cultured human peripheral blood mononuclear cells (PBMCs), this MEK inhibitor similarly suppressed TNF-α production by LPS. For the mechanism study, we found that trametinib blocked LPS-induced MEK-ERK activation in above monocytes, which accounted for the defective TNF-α response. Macrophages or PBMCs treated with a traditional MEK inhibitor PD98059 or infected with MEK1/2-shRNA lentivirus exhibited a similar defect as trametinib, and nullified the activity of trametinib. On the other hand, introducing a constitutively-active (CA) ERK1 restored TNF-α production by LPS in the presence of trametinib. In vivo, mice administrated with trametinib produced low levels of TNF-α after LPS stimulation, and these mice were protected from LPS-induced endotoxin shock. Together, these results show that trametinib inhibits LPS-induced TNF-α expression and endotoxin shock probably through blocking MEK-ERK signaling.

Protective effect of Auricularia auricular-judae polysaccharide on pulmo-nary tissues of rats with LPS-induced acute lung injury / 中国病理生理杂志

AIM:To investigate the effects of Auricularia auricular-judae polysaccharide ( AAP) on pulmonary tissues of rats with LPS-induced acute lung injury (ALI) and its mechanisms.METHODS: Adult Sprague-Dawley rats were randomly divided into control group , LPS group, low-dose AAP group, middle-dose AAP group, high-dose APP group, and dexamethasone group.The rats were injected with LPS (8 mg/kg, ip) to induce ALI.The rats in the AAP groups were treated with AAP for 7 d before the induction of ALI .The protein concentration in the bronchoalveolar lavage fluid (BALF) was measured.The lung edema degree was measured by detecting the wet /dry weight ratio.The myeloper-oxidase ( MPO) , total antioxidant capacity ( T-AOC) , total superoxide dismutase ( T-SOD) , nitric oxide synthase ( NOS) and malondialdehyde ( MDA) levels were determined .The pathological changes of the lung tissues were evaluated by HE staining.RESULTS:Treatment with AAP significantly improved LPS-induced lung pathological changes , attenuated the protein concentration in the BALF and wet/dry weight ratio , inhibited the activities of MPO and NOS , reduced MDA level and increased the activities of T-AOC and T-SOD.CONCLUSION: AAP protects against LPS-induced acute lung injury in rats.

Lentivirus mediated CCN1 gene on growth and migration of rat bone marrow mesenchymal stem cells / 中国病理生理杂志

[ABSTRACT]AIM:Toinvestigatetheroleofcysteine-rich61(Cyr61/CNN1)inproliferationandmigrationof bone marrow mesenchymal stem cells ( BMSCs ) .METHODS: The lentiviral vector carrying CCN 1 ( Lenti-GFP-CCN1 ) was constructed and then transfected into the rat BMSCs .The cells were divided into non-transfection group , transfection group ( transfected with Lenti-GFP-CCN1 ) and negative control group ( Lenti-GFP ) .The fluorescence intensity of the transfected BMSCs was observed under inverted fluorescence microscope .The effects of CCN1 on the proliferation and mi-gration of BMSCs were detected by MTT assay and scratch wound healing assay .RESULTS:The proliferation of BMSCs transfected with Lenti-GFP CCN1 had no significant difference compared with negative control group and control group .The width/thickness ratio of migrated BMSCs in wound healing was significantly higher in Lenti-GFP-CCN1 group than that in negative control group and control group (P<0.05).CONCLUSION:Exogenous CCN1 promotes the migration of BMSCs.

Recombinant adeno-associated virus serotype 9 with p65 ribozyme protects H9c2 cells from oxidative stress through inhibiting NF-κB signaling pathway / 老年心脏病学杂志（英文版）

Background Oxidative stress is a major mechanism underlying the pathogenesis of cardiovascular disease. It can trigger inflammatory cascades which are primarily mediated via nuclear factor-κB (NF-κB). The NF-κB transcription factor family includes several subunits (p50, p52, p65, c-Rel, and Rel B) that respond to myocardial ischemia. It has been proved that persistent myocyte NF-κB p65 activation in heart failure exacerbates cardiac remodeling. Mechods A recombinant adeno-associated virus serotype 9 carrying enhanced green fluorescent protein and anti-NF-κB p65 ribozyme (AAV9-R65-CMV-eGFP) was constructed. The cells were assessed by MTT assay, Annexin V–propidium iodide dual staining to study apoptosis. The expression of P65 and P50 were assessed by Western blot to investigate the under-lying molecular mechanisms. Results After stimulation with H2O2 for 6 h, H9c2 cells viability decreased significantly, a large fraction of cells underwent apoptosis. We observed a rescue of H9c2 cells from H2O2-induced apoptosis in pretreatment with AAV9-R65-CMV-eGFP. Moreover, AAV9-R65-CMV-eGFP decreased H2O2-induced P65 expression. Conclusions AAV9-R65-CMV-eGFP protects H9c2 cells from oxidative stress induced apoptosis through down-regulation of P65 expression. These observations indicate that AAV9-R65-CMV-eGFP has the potential to exert cardioprotective effects against oxidative stress, which might be of great importance to clinical efficacy for cardiovascular disease.

Misdiagnosis of aortic dissection: experience of 361 patients.

Aortic dissection (AD) is a life-threatening condition that requires immediate diagnosis and surgical correction. Patients with acute AD usually present clinically with an insignificant medical history, leading to a high probability of misdiagnosis. The aim of the present study was to investigate the number of misdiagnoses of patients with AD in order to understand this problem and to avoid future misdiagnosis in the emergency department. Clinical data from 361 patients with AD admitted between January 2003 and June 2008 were reviewed as part of a retrospective chart review. Diagnosis of AD was made using either chest x-ray, computed tomography, magnetic resonance imaging, or angiography. Fifty-one patients had an initial misdiagnosis (14.1%), later found to have experienced AD. The condition may clinically present in a varied number of manifestations, including syncope, chest pain, abdominal pain, back pain, acute congestive heart failure, or alternatively with minimal symptoms. Persons of any age can experience an AD, with key clinical manifestations of pain. Echocardiography can be used for primary examination of patients with suspected AD; however, a definite diagnosis is usually made using computed tomographic or magnetic resonance angiography. Care should be taken, particularly in the emergency department, to maintain a level of suspicion for AD diagnosis in order to avoid the potential for misdiagnosis.

Analysis of risk factors of operation on 185 patients with acute type A aortic dissection / 中华急诊医学杂志

Objective To assess the risk factors of the in-hospital mortality of acute type A aortic dissection after operation. Method From January 2003 to June 2008,185 patients, 144 males and 41 females, with acute type A aortic dissection operated on were enrolled. The average age of patients was (49.46 ± 11.04 ) years old.The patients' demographics, history, clinical features, and some laboratory examinations were reviewed. Univariate and multivariate analysis followed by logistic regression analysis were carried out to identify the predictors of inhospital mortality. Results The in-hospital mortality rate was 9.1%. The results of univariate and multivariate analyses as follows: pre-operation positive neurological symptom (Univariate OR = 5.084,95%CI:1.792 -14.426, P = 0.002; Multivariate OR = 5.538,95%CI: 1.834 - 16.721, P = 0.002, respectively), hypotension (Univariate OR = 6.986,95%CI:1.510- 32.323,P =0.013; multivariate OR = 1.998,95%CI:0.315-12.679,P = 0.463, respectively) and renal failure (Univariate OR = 3.594,95%CI:1.237 - 10.438,P =0.019; Multivariate OR = 3.254,95%CI:1.034- 10.242, P= 0.044, respectively). Conclusions There are two predictors, pre-operation positive neurological symptom and renal failure, of pre-hospital mortality found in current analyses. Our results may improve the regimen made by cardiac surgeons and emergency doctors so as to help patients and their relatives to make correct decision.

Association study of TLR4 polymorphisms with severe community-acquired pneumonia susceptibility and outcome / 中华急诊医学杂志

Objective To investigate the possible association of TLR4 polymorphisms with susceptibility and prognosis of SCAP.Method A total of 360 CAP patients hospitalized in emergency department of Zhongshan hospital from May 2005 to April 2008 were enrolled in this case-control study.Patients were excluded if they had metastatic tumors,autoimmune diseases,AIDS or received immunosuppressive drugs.This study was approved by the ethical committee of Zhongshan hospital,Fudan University.Patients were divided into SCAP group(n = 180)and NSCAP group(n = 180)according to the illness severity,and were divided into survival group(n = 300)and death group(n = 60)according to the 30-day mortality.Hapmap database of Han Chinese population was used to select the Tag SNPs.Primer 3 software was used to design the PCR and sequencing primers.Genomic DNA was extracted from peripheral blood mononuclear cells.Genotyping was performed by sequencing the PCR products.We used X2 analysis to evaluate the significance of differences in genotype and allele frequencies between different groups.Results The distributions of three TagSNPs(rs2149356,rs11536879,rs1927907)were consistent with Hardy-Weinberg equilibrium.The allele and genotype frequencies of three TagSNPs in the SCAP group did not differ from the NSCAP group.Also,no significant difference was found between survivor group and non-survivor group.The haplotype frequencies of CA,TA and TG were not significantly different between SCAP group and NSCAP group.And no significant difference of haplotype frequency was existed between survivor group and nonsurvivor group.Conclusions This study suggested that TLR4 gene polymorphisms were not significantly associated with the susceptibility and prognosis of SCAP.

Change of the serum pro-and anti-inflammatory cytokines in sepsis patients / 中华急诊医学杂志

Objective To investigate the characteristics of inanune status change in sepsis and severe sepsis patients by quantitative analysing the serum concertrations of pro-and anti-inflammatory cytokines. Method Serum of 38 sepsis patients, 32 severe sepsis patients were collected and 15 health individuals were as controls.ELJSA method was used to quantify the serum levels of inflammatory cytokines. The severity of patient's condition was assessed according to the APACHE Ⅱ system, Retolts The serum concentrations of inflammatory cytokines were different among sepsis and severe sepsis patients. In the serum of sepsis patients the pro-inflammatory eytokine were dominant. But in the serum of severe sepsis patients the anti-inflammatory cytokine were dominant.The serum levels of TNF-α, IL-6, IL-1, IL-10 were obviously different among control, sepsis and severe sepsis groups ( P<0.05). The serum levels of IL-1 and IL-10 were significantly correlated with APACHE II scores. The multiple linear regression eqution was APACHE Ⅱ=- 9.393 + (IL-10 x 0.550) + (IL-1 x 0.305) (F =26.198,P<0.001) Conclusions The serum levels of pro-and anti-inflammatory cytokines were significantly different among patients with different stages of sepsis, and the immune status were different. The activation of inflatrmmtory reaction were constant in sepsis patients, while the expression of anti-inflammatory cytokines increased in severe sepsis patients.