Aim To study the role of TNF-α/NF-κB signaling in matrix metalloproteinase ( MMP)-9 expres-sion induced by lipopolysaccharide ( LPS ) in airway epithelial cells, and to investigate the effects of lenti-virus mediated RNAi targeting a disintegrin and metal-loproteinase 17 ( ADAM17 ) gene on MMP-9 expression induced by LPS. Methods The ADAM17 siRNA ex-pression vector was constructed, and packaged to re-combinant lentivirus in 293T cells. The HBE4-E6/E7 cells were pretreated for 30 min by NF-κB inhibitor ( PDTC) and a recombinant human TNFR p75-Fc fu-sion protein ( Etanercept) , or infected by the recombi-nant lentivirus for 72 h, and then stimulated for 24 h by LPS or TNF-α. The release of TNF-α was detected by ELISA. The mRNA and protein levels of MMP-9 were analyzed respectively by RT-PCR and Western blot. NF-κB activity was detected by electrophoretic mobility shift assay. Results LPS and TNF-α signifi-cantly increased MMP-9 mRNA and protein expressions and the activation of NF-κB in HBE4-E6/E7 cells ( P<0. 05 ) . Etanercept and PDTC significantly inhibited MMP-9 expression and the activation of NF-κB induced by LPS ( P<0. 05 ) . Lentivirus mediated RNAi targe-ting ADAM17 significantly decreased TNF-α produc-tion, inhibited MMP-9 mRNA and protein expressions and the activation of NF-κB induced by LPS in HBE4-E6/E7 cells ( P <0. 05 ) . Lentivirus mediated RNAi targeting ADAM17 did not inhibit MMP-9 mRNA and protein expressions and the activation of NF-κB in-duced by TNF-α ( P>0. 05 ) . And PDTC significantly inhibited MMP-9 mRNA and protein expressions and the activation of NF-κB induced by TNF-α ( P <0. 05 ) . Conclusions TNF-α/NF-κB signaling partic-ipates in the regulation of MMP-9 expression induced by LPS in airway epithelial cells, and lentivirus-media-ted RNAi targeting ADAM17 plays an important role in that signaling pathway upstream by regulating TNF-αrelease.

Objective To investigate the effect and mechanism of phosphorylated protein kinase R-like ER kinase(p-PERK) and C/EBP homologous protein(CHOP) after hypoxic-ischemic brain damage ( HIBD) . Methods Neonatal 7-day-old Sprague Dawley rats were divided into sham-operation control group and HIBD group( n=30 per group) . Each group was divided into 0 h,6 h and 24 h subgroup after operation ( n=10 per group) . The ratio of apoptosis of brain cell was measured by flow cytometer and the expression of p-PERK and CHOP were detected by Western blot. Results (1)Apoptosis cell appeared at 6 h in HIBD group,the ratio of cell apoptosis was(2. 17 ± 0. 19)%. The apoptosis cell obvious increased at 24 h,the ratio of cell apoptosis was(13. 42 ± 0. 83)%. There was a significant increase in the ratio of apoptosis after HIBD 6 h and 24 h, as compared with sham-operation control group [ ( 0. 57 ± 0. 06 )%( P <0. 01 ) ] . ( 2 ) The expression of both p-PERK and CHOP was very low in sham-operation control group. In the HIBD group,the expression of both p-PERK and CHOP began to increase at 6 h and increased furthermore at HIBD 24 h. The differences in the expression levels of p-PERK and CHOP in HIBD group among different time points were significant( P<0. 01 ) . ( 3 ) The expression of p-PERK positively correlated with the expression of CHOP (r=0. 997,P< 0. 05). Conclusion With the emerging of apoptosis after HIBD,the expression of both p-PERK and CHOP increases. The imbalance in the expression of PERK induces the apoptosis of brain cells in the HIBD of neonatal rats by regulation of CHOP expression.

Objective The discovery of microRNA (miRNA) in maternal serum has opened up new possibilities for non-invasive prenatal diagnosis.However,our understanding of these pregnancy-related miRNA in the serum of pregnant women with fetuses with neural tube defects (NTDs) is still limited.This article is to study the dysregulated expression of microRNA-423 (miR-423) in the serum of pregnant women with neural tube defect(NTD) fetuses and its potential role as a biomarker for non-invasive prenatal diagnosis of fetal NTD.Methods Thirty-three pregnant women whose fetuses were diagnosed as neural tube defects by ultrasound (22 cases of spina bifida and 11 cases of anencephaly)and 33 normal pregnant women were selected.Peripheral venous blood of each pregnant woman was obtained early in the morning,the serum was purified from blood by centrifugation,then total RNA was isolated from serum and the miR-423 levels were detected by real-time RT-PCR.The ROC curve was used for assessing the diagnostic accuracy of miR-423 for fetal NTD.Results We revealed miR-423 with signifcant down-regulation in expression in serum of pregnant women with NTD fetuses (0.96 ±0.14) compared as women with normal pregnancies(2.28 ±0.43) (P ＜0.05).We performed ROC analysis of data from the 33 case-control pairs.The expression of miR-423 could distinguish NTD cases from normal controls,with an AUC of 0.711 (95 ％ CI:0.566 ～ 0.856) (P ＜ 0.05).Moreover,the expression of miR423 decreased only in serum of pregnant women with anencephaly fetuses(0.58 ±0.08)by the analysis in different forms of NTD.Conclusion miR-423 is deregulated in the serum of pregnant women with NTD fetuses and highlight the clinical potential of miR-423 as biomarker for diagnosis and prognostication of fetal NTD.

Agmatine ; administration & dosage ; pharmacology ; Analgesia ; methods ; Animals ; Injections, Spinal ; Male ; Morphine ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Spinal Cord ; drug effects

Objective To establish mild cognitive dysfunction (MCI) models in elderly rats,and to investigate the pathophysiological features.Methods Totally 40 SD rats (14 to 18-month-old) were randomly divided into 2 groups:the model group (n=20) and the sham operation group (n=20).Bilateral carotid artery stenosis was prepared in the model group while bilateral carotid artery was seperated with no bilateral narrowing in the sham operation group.30 days after the operation,Morris water maze test was performed,pathomorphological and electron microscopic observations of the cerebral tissue were examined and the expression of G protein-coupled receptor kinase 2(GRK2) in hippocampus tissue w detected by reverse transcription polymerase chain reaction (RT-PCR) and Western blottin.Results The mortality in model group was only 10％.Pathological morphology and ultrastructure showed that hippocampal tissue structure was almost normal in sham operated group,but in model group group,hippocampal CA1 pyramidal cells were in ischemic demyelination,arranged loose,and part of the cells showed nucleus pyknosis,deeply stained; there was no obvious infarct in white matter,part of the white matter fiher hecame thinner and disorder,nucleolus became smaller and steped aside,cytoplasmic electron density increased,lipofuscin appeared occasionally.Rough endoplasmic reticulum and Golgi were expanded,cytosolic free ribosomes increased,part of mitochondria became swelled,vacuolated.Morris water maze test results showed that the average escape latency in model group was longer than in sham group (P＜0.05).In spatial probe test,the average time of crossing the first original platform in model rats was significantly longer than the sham operated group [(36.80±7.68) s vs.(20.87±6.16)s,P＜0.05].The average number of crossing the original platform in 60 seconds in model group was significantly less than in sham group(1.43±0.51 vs.3.10±1.45,P＜0.05).The expressiones of GRK2 mRNA and protein in the hippocampus were significantly increased in model group rats than in sham group (P＜0.05).Conclusions The model of severe CCA stenosis in elderly rats can be applied for MCI animal models with good stability and repeatability.Compared with sham group,the cells morphology and ultrastructure in model group appeare more obvious pathological changes and mild impairments in cognitive function.GRK2 may play an important role in the development of MCI.

Objective To identify two suspected Legionella pneumophila (L.pneumophila) strains isolated from environmental water and air conditioning cooling water systems in Guangzhou city.Methods The two strains were identified by their cultural characteristics,biochemical test,Legionella-specific primer PCR identification,PCR-enzymatic digestion analysis,16S rRNA,mip and rpoB gene sequencing analysis.Results The two suspected L.pneumophila isolates were identified as gram-negative bacillus appeared as white colonies on BCYEα-agar after incubation for 48 hours at 36℃.Both isolates were positive for oxidase,gelatinase and hydrolysis of hippurate,and negative for urease activity and nitrate reduction.Their phenotypic characteristics were similar to those of L.pneumophila strains.Results of PCR identification by using Legionella-specific primer were positive.Enzymatic digestion analysis showed that the 226 bp PCR products of two isolates were not digested by Taa Ⅰ.The two strains were classified as Legionella busanensis as indicated by gene sequencing analysis of 16S rRNA,mip and rpoB gene.Conclusion Two L.busanensis strains were first isolated from environmental and air conditioning cooling water systems in China.Due to their biochemical characteristics,L.busanensis strains were commonly misidentified as L.pneumophila,but could be effectively identified by PCR-enzymatic digestion analysis and multiple genes identification.

Objective To observe the effects of a rehabilitation robot and body weight supported treadmill training (BWSTT) on the walking ability of hemiplegic patients after stroke.Methods Sixty hemiplegic patients were divided randomly into treatment and control groups (n=30 in each).The treatment group received training assisted by a leg rehabilitation robot for 10-20 min once daily,6 days per week for 8 weeks in addition to conventional treatments and BWSTT.The control group was only given conventional treatments and BWSTT once daily for 8 weeks.Their lower extremity functions,balance and walking ability were assessed with a simplified Fugl-Meyer assessment (FMA) and using the Berg balance scale (BBS) and Functional Ambulation Categories (FACs).Results After training,both groups showed significant improvements in terms of FMA,BBS and FAC results.The treatment group showed significantly better improvements in lower extremity function,balance and walking ability compared with the control group.Conclusions The lower-limb rehabilitation robot and BWSTT could together improve balance and walking ability.

Objective To investigate the association between nutrition and peripheral artery disease (PAD) in continuous ambulatory peritoneal dialysis (CAPD) patients.Methods One hundred and two stable CAPD patients from a single center were enrolled in this cross-sectional study.Automatic ankle-brachial index (ABI) measuring system was applied to examine ABI.Patients were divided into PAD group (ABI ＜ 0.9) and non-PAD group (ABI≥0.9).Clinical data were collected.Biochemical parameters were detected.Nutritional status was evaluated by serum albumin,handgrip strength (HGS) and subjective global assessment (SGA).Logistic regression analysis was performed to estimate the association of PAD with each nutritional marker as well as other potential risk factors.Results The incidence of PAD was 23.53％ (24/102).ABI was significantly lower in patients with malnutrition as compared to those without malnutrition [(0.72 ± 0.21) vs (1.04 ± 0.14),P＜0.01].Compared with non-PAD patients,serum albumin (P ＜ 0.01),HGS (P ＜ 0.01),diastolic blood pressure (P ＜ 0.05),serum creatine (P ＜ 0.05)、blood urine nitrogen (P ＜ 0.01) were significantly decreased,but age (P ＜ 0.01),the incidence of malnutrition [SGA,P ＜ 0.01],diabetic status (P ＜ 0.01),cardiovascular disease history (P ＜ 0.01) were significantly increased in PAD patients.Logistic regression analysis showed that serum albumin (OR=0.762,95％ CI:0.611-0.948,P=0.015),HGS (OR=0.988,95％ CI:0.979-0.997,P=0.013) were independent protective factors for PAD,malnutrition [(SGA),OR=21.101,95％ CI:5.008-88.901,P＜0.01] was independent risk factor for PAD in CAPD patients.Conclusions The PAD incidence of CAPD patients in our center is 23.53％.Nutrition is independent factor associated with PAD in CAPD patients.

Objective To investigate the causes of arterial reocclusion in diabetic feet patients after endovascular treatment and its remedial measures.Methods From January 2009 to October 2013,clinical data of 371 arterial reocclusion of diabetic feet patients after endovascular treatment in Tianjin First Central Hospital were reviewed retrospectively.We summarized the causes of reocclusion,treatment methods and the short term results.Results According to the Trans-Alantic Inter-Society Consensus (TASC) Ⅱ grading standards,the first time when the endovascular treatment started there were 37 cases of grade A,85 cases of grade B,143 cases of grade C,106 cases of grade D.Arterial re-occlusion developed from one day to 36 months,averaging at (21 ± 8) months.Causes of re-occlusion included intimal hyperplasia in 263 cases (70.9％),thrombosis in 65 cases (17.5％),dissection in 19 cases (5.1％),stent fracture in 17 cases (4.6％),vascular rupture in 7 cases (1.9％).Remedial therapy adopted for arterial reocclusion was repeated endovascular treatment in 327 cases (88.1％),arterial bypass surgery in 23 cases (6.2％),conservative treatment in 13 cases (3.5％),amputation (cut toe) in 4 cases (1.1％),4 cases (1.1％) died perioperatively.275 cases were followed up for 1 to 36 months,the average was (13 ± 8) months.patency rate was 82.9％,71.3％ and 63.0％ at 6 months,1 year and 2 years.Amputation rate was 1.1％,1.8％ and 2.5％ at 6 months,1 year and 2 years.Conclusions Intimal hyperplasia is to blame for arterial reocclusion after endovascular treatment of diabetic foot.In this case most patients still can benefit from second time endovascular treatment,with a satisfactory short term patency rate.

OBJECTIVE:To provide reference for rational use of TCM injections. METHODS:Among the inpatients who used TCM injections in 4 hospitals of Shanghai Jiading district in 2013,100 cases were collected by systematic sampling in each hospital separately, and the data were analyzed retrospectively. RESULTS:The varieties and amount of TCM injections for blood-activating and stasis-resolving both occupied the first place. 22% inpatients used two or more TCM injections at the same time. The proportion of overdose use of TCM injections reached 54.55%. The type of unreasonable drug use mainly included drug use against differentiation of syndromes,super indications drugs and solvent error. Combined use of TCM injections with same ef-fect accounted for 20.45% of combined use of TCM injections. CONCLUSIONS:It is effective means to improve rational drug use that great impertance paid for clinical use of TCM injections.