Objective:To detect the effect of microRNA-7 ( miR-7 ) knockdown on pathology in murine acute lung injury ( ALI) model,and preliminarily explore its significance.Methods:Murine ALI model was performed by intraperitoneal injection of Li-popolysaccharide (LPS) (10 mg/kg) into miR-7KD mice and wild-type (wild type,WT) mice respectively.Then,the pathologic injury of lung tissue were observed by HE staining.And total cell count of bronchoalveolarlavage(BAL) was calculated.The relative expression of related cytokines in lung tissue was analyzed by Real-time PCR assay.Furthermore,the changes on proportion of innate immune cells (γδT cell and F4/80 macrophages cell) and adaptive immune cell ( CD4+T cell and CD8+T cell) were analyzed by FACS.Meanwhile, the expression of CD62L and CD69,as well as the absolute number,in CD4+T cell were also analyzed.Results: Compared with WT mice,pathological damage in lung tissues was significantly alleviated in miR-7KD mice.Real-time PCR analysis showed that the relative expression of IL-6 was obviously reduced (P<0.01),conversely,relative expression of IL-4 and TGF-βwere obviously increased (P<0.05).Furthermore,the total cell number in BAL also reduced significantly (P<0.05).Importantly,FACS analysis showed that the proportion and the absolute number of F4/80+Mφcells obviously reduced (P<0.05);however,the proportion of γδT cells increased (P<0.05).Moreover,the proportion and the absolute number of CD4+T cells and CD8+T cells were significantly reduced (P<0.05). Finally, the proportion and the absolute number of CD62L+in CD4+T cells were upregulated vigorously,contrastly,the proportion and the absolute number of CD69+in CD4+T cells were notably up-regulated (P<0.05).Conclusion:miR-7 defeciency could significantly ameliorate the pathology of murine ALI,suggesting that it may play an important regulatory role in the development of ALI.

BACKGROUNDPulmonary embolism (PE) can be difficult to diagnose in elderly patients because of the coexistent diseases and the combination of drugs that they have taken. We aimed to compare the clinical diagnostic values of the Wells score, the revised Geneva score and each of them combined with D-dimer for suspected PE in elderly patients.

METHODSThree hundred and thirty-six patients who were admitted for suspected PE were enrolled retrospectively and divided into two groups based on age (≥65 or <65 years old). The Wells and revised Geneva scores were applied to evaluate the clinical probability of PE, and the positive predictive values of both scores were calculated using computed tomography pulmonary arteriography as a gold standard; overall accuracy was evaluated by the area under the curve (AUC) of receiver operator characteristic curve; the negative predictive values of D-dimer, the Wells score combined with D-dimer, and the revised Geneva score combined with D-dimer were calculated.

RESULTSNinety-six cases (28.6%) were definitely diagnosed as PE among 336 cases, among them 56 cases (58.3%) were ≥65 years old. The positive predictive values of Wells and revised Geneva scores were 65.8% and 32.4%, respectively (P < 0.05) in the elderly patients; the AUC for the Wells score and the revised Geneva score in elderly was 0.682 (95% confidence interval [CI]: 0.612-0.746) and 0.655 (95% CI: 0.584-0.722), respectively (P = 0.389). The negative predictive values of D-dimer, the Wells score combined with D-dimer, and the revised Geneva score combined with D-dimer were 93.7%, 100%, and 100% in the elderly, respectively.

CONCLUSIONSThe diagnostic value of the Wells score was higher than the revised Geneva score for the elderly cases with suspected PE. The combination of either the Wells score or the revised Geneva score with a normal D-dimer concentration is a safe strategy to rule out PE.

Aged ; Aged, 80 and over ; Angiography ; Female ; Fibrin Fibrinogen Degradation Products ; metabolism ; Humans ; Male ; Pulmonary Embolism ; diagnosis ; diagnostic imaging ; metabolism ; Retrospective Studies

Objective To analyze the surveillance results and grasp the situation of Keshan disease in Wudalianchi city Heilongjiang province.Methods In 2009,Kaifa village was selected as the surveillance point in Wudalianchi city,total resident population were monitored by routine clinical examination and 12-lead electrocardiogram(ECG) tracing.Suspected cases with Keshan disease were taken chest X-ray,and Keshan disease was diagnosed based on Keshan Disease Diagnostic Criteria (WS/T 210-2011).Results A total of 795 people were investigated,including 397 males and 398 females.Eighteen people were found to be the patients with Keshan disease,of which 13 cases were latent Keshan patients,5 cases were chronic Keshan patients.The overall detection rate was 2.27％,aged 24 to 83 years old.There was no acute type and subacute type of Keshan disease in the surveillance point.Twenty nine cases of abnormal ECG were detected,the detection rate was 3.65％ (29/795),of which the 18 patients with Keshan disease were all had abnormal ECGs,mainly taken the form of ST-T changes and completely right bundle branch blocked.Six cases of male patients with Keshan disease were detected,the detection rate was 1.52％ (6/397); 12 cases of female patients with Keshan disease were detected,the detection rate was 3.01％ (12/398).Conclusions There is still potential and chronic Keshan disease cases in Wudalianchi city.We must keep on the monitoring on Keshan disease,master the dynamical changes of the disease conditions,and carry out the targeted prevention and control of Keshan disease.

Objective To investigate the prevalence and incidence of Keshan disease (KD) and the selenium concentration of food and hair in residents of Yongjin Village, Fuyu County, Heilongjiang Province, national monitoring site, in 2007. Methods According to the Standard of Keshan Disease Surveillance and the Standard of Diagnosis of Keshan Disease(GB 17021-1997), the residents living in the monitoring site were surveyed by clinical examination and electrocardiography. For individuals whose hearts showed abnormalities, a chest X-ray photograph was taken. The selenium concentrations of the residents' food (flour) and hair were assayed by flowing injection hydride generation atomic fluoremetric method(FI-HG-AFM). Results Nineteen KD patients were found from 282 residents in 2007 KD surveillance. The prevalence of KD, latent KD and chronic KD were 6.7%(19/282), 2.8%(8/282) and 3.9%(11/282), respectively. Five of the 8 latent KD cases were newly found. In addition, there were 5 the suspected KD cases, including 2 suspected chronic KD cases. No acute KD or sub-acute KD patients were found in Yongjin Village at this monitoring site this year. The average selenium concentration of children hair and residents food were (0.3197±0.0586)mg/kg and (0.0210±0.0062)mg/kg, respectively. Conclusions New cases of KD continued to emerge, indicating that etiological factors still exist. Therefore, the emphasis of monitoring KD in furore is founding the consummate report of infectious disease system and training the personnel to increase the reliability of monitoring.

AIMTo study the effects of TNF receptor blocking peptide on adjuvant arthritis in rats.

METHODSThe model of rat adjuvant arthritis was induced by injection of complete Freund's adjuvant. The TNF receptor blocking peptide was injected locally in the ankle. The ankle swelling, the pathologic changes in the ankle joint and the expression of IL-1 beta mRNA and TNF-alpha mRNA by peritoneal macrophages (RT-PCR) were observed.

RESULTSThe model of rat adjuvant arthritis induced by injection of complete Freund's adjuvant was similar to human rheumatoid arthritis. The treatment with TNF receptor blocking peptide for 10 days resulted in complete inhibition of joint swelling, a decrease in infiltration of inflammatory cell into joint tissue, an obvious alleviation of inflammatory pathological damages and an apparent decline of TNF-alpha mRNA and IL-1 beta mRNA of peritoneal macrophages of rats.

CONCLUSIONThe TNF receptor blocking peptide can protect the joint from inflammatory damage induced by adjuvant arthritis by suppression of TNF-alpha and IL-1 production, thereby alleviating the pathological injury of joint and controlling effectively the clinic course of arthritis.

Animals ; Ankle Joint ; pathology ; Arthritis, Experimental ; immunology ; pathology ; Interleukin-1 ; biosynthesis ; genetics ; Macrophages, Peritoneal ; metabolism ; Male ; Peptides ; pharmacology ; RNA, Messenger ; genetics ; Rats ; Rats, Wistar ; Receptors, Tumor Necrosis Factor ; antagonists & inhibitors ; Tumor Necrosis Factor-alpha ; biosynthesis ; genetics

OBJECTIVE Glioblastoma multiforme (GBM) is the most malignant primary tumor of the central nervous system and is associated with a very poor prognosis. No further improvements in outcomes have been reported since radiotherapy-temozolomide therapy was introduced.Therefore,de-veloping new agents to treat GBM is important. This study aimed to evaluate the anti-tumor effect of evodiamine (Evo) on GBM cells, and to determine the underlying mechanisms involved. METHODS U251,LN229,HEB and PC12 cells were treated with various concentrations of evodiamine for 24 and 48 hours,cell viability was measured by MTT assay.The U251 and LN229 cells were treated with evo-diamine(0-10 μmol·L-1)for 24 h,and then stained with Hoechst 33258.An Annexin V-FITC Apoptosis Detection Kit was used to detect apoptosis in the cells.Reactive oxygen species(ROS)production was detected using dichlorofluorescein diacetate (DCFH-DA) staining. The changes in mitochondrial mem-brane potential (MMP) were assessed by JC-1 after cells were treated with evodiamine. The expres-sion levels of p-PI3K,PI3K,p-Akt,Akt,Bax,Bcl-2,p-p38,p38,p-JNK,JNK,p-ERK,ERK,Cytochrome c, Caspase-3, cleaved Caspase-3, PRAP, and cleaved PARP were measured by Western blot analy-ses. RESULTS According to MTT assay results, Evo significantly inhibited the cell proliferation in a time- and dose-dependent manner. Fluorescence microscopy and flow cytometry analyses revealed that Evo induced cell apoptosis in a concentration-dependent manner.Moreover,Evo induced reactive oxygen species (ROS) production and mitochondrial membrane potential (MMP) disruption. Finally, Evo induced apoptosis in cancer cells by suppressing PI3K/AKT signaling and inducing MAPK phos-phorylation(p38 and JNK,but not ERK)to regulate apoptotic proteins(Bax,Bcl-2,Cytochrome c,Cas-pase-3, and PARP). CONCLUSION In summary, Evo inhibits cell proliferation by inducing cellular apoptosis via suppressing PI3K/AKT and activating MAPK in GBM;these results indicate that Evo may be regarded as a new approach for GBM treatment.

OBJECTIVETo investigate the effect of astragalus (As) on calcium accumulation and SERCA2a gene expression in left ventricular tissues in rats with pressure overload-induced cardiac hypertrophy.

METHODcardiac hypertrophy was induced by clipping the abdominal aorta in rats. Male SD rats were allocated to six groups: sham-operrated (Sham), aortic stenosis (Model), model +As-L (5 g x kg(-1) x d(-1)), model+As-M (10 g x kg(-1) x d(-1)), model+As-H (20 g x kg(-1) x d(-1)) and model + captopril (0.05 mg x kg(-1) x d(-1), a positive control). The drugs were administered orally from the 13 th week after surgery. Rats were examined after 12 week treatment with drugs. The cardiac hypertrophy was evaluated by left ventricular mass index (LVMI, left ventricular weight/ body weight). The calcium content in left ventricular tissue was measured by atomic absorption spectrometry. SERCA2a mRNA and protein expressions in left ventricular tissues were determined by half-quantitative RT-PCR and Western blot normalized to abundance of GAPDH mRNA and protein, respectively.

RESULTThe increase of LVMI was dose-dependently lessened by As (P < 0.01, P < 0.001). The effect of As-H was similar to that of Captopril. As markedly attenuated calcium accumulation in myocardial tissure (P < 0.01). RT-PCR and Western blot results demonstrated that SERCA2a gene expressions were downregulated (P < 0.05) significantly in model group compared with sham group. As-H upregulated SERCA2a gene expressions (P < 0.05), whereas Captopril had no effect on that.

CONCLUSIONThe inhibition of As on left ventricular hypertrophy induced by pressure overload in rats may partly contribute to its attenuation of calcium accumulation and up-regulation of SERCA2a gene expressions in left ventricular tissues.

Animals ; Astragalus Plant ; chemistry ; Blotting, Western ; Calcium ; metabolism ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; Gene Expression Regulation ; drug effects ; Heart ; drug effects ; Hypertrophy, Left Ventricular ; metabolism ; physiopathology ; Male ; Myocardium ; metabolism ; Rats ; Rats, Sprague-Dawley ; Reverse Transcriptase Polymerase Chain Reaction ; Sarcoplasmic Reticulum Calcium-Transporting ATPases ; genetics ; metabolism

Objective To study the relationship between Couagen Ⅰ,MMP-2,TIMP-2 gene expression and atrial fibrosis during heart failure(HF)in dog.Methods Fourteen dogs were used and randomized into HF induced by ventricular tachypacing and control group.Burst atrial pacing was used to induce atrial fibrillation(AF).And the mRNA and protein level of collagen Ⅰ,MMP-2 and TIMP-2 were detected by RT-PCR and immunohistochemical technique.Tissue samples were stained with Mallory trichrome.Results Left ventricular ejection fraction (LVEF) decreased from (67.4? 6.0)% to (29.2?7.8)%,the inducible rate of AF(7/7 vs 2/7) and sustained AF(5/7 vs 0/7) increased and duration of AF stabeatrial fibrillation(SAF) [(462.12?181.43)s vs(0.57?0.57) s] prolonged significantly in HF group.Atrial fibrous tissue content and atrial size of HF group were significantly greater than the controls dogs(268.8% in lefe atria and 190.3% in right atria).The mRNA and protein level of collagen Ⅰ(56.2% and 132.2% in lefe atria,37.4% and 78.0% in right atria)and MMP-2 (100.0% and 115.7% in lefe atria,65.7% and 96.8% in right atria) increased evidently in both lefe atria and right atria,TIMP-2 mRNA decreased 46.3% in lefe atria and had no change in right atria and that its protein had no change in both atrium,whereas the ratio of MMP-2/ TIMP-2 of mRNA and protein increased markedly in both lefe atria (285.3% and 148.8%)and right atria (106.1% and 134.7%)of HF group.SAF had a positive correlation with fibrosis and the gene level of collagen Ⅰ in lefe atria,the ratio of MMP-2/TIMP-2 had a positive correlation with fibrosis and collagen Ⅰ gene level in lefe atria during HF.Conclusions The changes of collagen Ⅰ,MMP-2 and TIMP-2 gene expression appear to be a molecular mechanism of AF, and the molecular remodeling of collagen Ⅰ induced by regulation unbalance of MMP-2/TIMP-2 appears to be an important mechanism of atrial fibrosis during HF.

Aged ; Humans ; Male ; Urinary Diversion ; adverse effects ; Urinary Fistula ; diagnosis ; etiology

OBJECTIVETo investigate the effects of ischemic postconditioning (IPostC) on pneumocyte apoptosis after lung ischemia/reperfusion injury in rats.

METHODSAdult male SD rats were randomly divided into 3 groups based upon the intervention (n = 8): control group (C), lung ischemic reperfusion group (LIR), LIR+ IPostC group (IPostC). At the end of the experiment, blood specimens drawn from the arteria carotis were tested for the content of malondialdehyde (MDA), the activity of superoxide dismutase (SOD) and myeloperoxidase (MPO); the pneumocyte apoptosis index (AI) was achieved by tennrminal deoxynucleotidyl transferase mediated dUTP nick end abeling (TUNEL); the expression of Bcl-2, Bax protein in lung tissue was accessed by quantitative immunohistochemistry (MHC) and Bcl-2, Bax mRNA by RT-PCR.

RESULTSIPostC could significantly attenuate the MDA level, MPO activity and improve SOD activity in blood serum which was comparable to I/R and significantly reduced the number of TUNEL-positive cells compared with I/R group, expressed as Al (% total nuclei) from (39.0 +/- 3.46) to (8.0 +/- 0.88) (P < 0.01). The protein and mRNA expression of Bcl-2 and Bax showed that IPO significantly attenuated the ischemia/reperfusion-upregulated expression of Bax protein but improved the expression of Bcl-2 that improved the Bcl-2/Bax ratio (P < 0.01) .

CONCLUSIONIPostC may attenuate pneumocyte apoptosis in LIRI by up-regulating expression of Bcl-2/Bax ratio and by inhibiting oxidant generation and neutrophils filtration.

Alveolar Epithelial Cells ; cytology ; Animals ; Apoptosis ; Ischemic Postconditioning ; Lung ; metabolism ; pathology ; Lung Injury ; physiopathology ; Male ; Malondialdehyde ; metabolism ; Peroxidase ; metabolism ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; physiopathology ; Superoxide Dismutase ; metabolism ; bcl-2-Associated X Protein ; metabolism