AIM:To study the feasibility of recombinant adeno-associated virus(rAAV)as a vector to transfer the green fluorescent protein(GFP)gene as a target gene into rabbit retina.METHODS:Intravitreal injection of rAAV-gfp was performed in either eye for each rabbit with the other eye taken as control.At the 3rd,7th,and 14th day after injection,the eyeballs were removed,and the retinas were flat-mounted on glass slides to inspect the retinal fluorescence,respectively.RESULTS:After intravitreal injection of rAAV-gfp,the presence of fluorescent spots in the cytoplasm of retinal cells indicated that GFP gene was efficiently transferred and expressed in the rabbit retina.CONCLUSION:Recombinant adeno-associated virus is a reliable and simple vector for transferring target gene,e.g.,GFP gene,to the retina.

Objective To study the expression level of peptidylarginine deiminase 4 (PADI4) and protein tyrosine phosphatase nonreceptor type 22 (PTPN22) in the synovium of rat model of collagen-induced arthritis, and to explore their possible therapeutic role in rheumatoid arthritis.
Methods Thirty-two female Wistar rats weighing 100±20 g were randomly assigned into 3-week collagen-induced arthritis (CIA) model group (n=8), 4-week CIA model group (n=8), 6-week CIA model group (n=8), and the control group (n=8). The body weight changes of each group were recorded. The expression levels of PADI4 and PTPN22 were detected and compared by the methods of immunohistochemical staining and Western blot.
Results Arthritis of rat began to form 14 days after sensitization and the joint swelling reached peak at 28 days. The weights of the rats slowly grew both in CIA model groups and the control group. Immunohistochemical staining results showed that the positive expression of PADI4 and PTPN22 was mainly located in cartilage peripheral mononuclear cells, the cytoplasm of infiltrated cells, and bone marrow cavity. There were significant differences in the optical density of PADI4 and PTPN22 among CIA model groups and the control group (PADI4, 0.2898±0.012, 0.2982±0.022, 0.2974±0.031, 0.2530±0.013 in 3-week CIA model, 4-week CIA model, 6-week CIA model and control groups;PTPN22, 0.2723±0.004, 0.2781±0.010, 0.2767±0.008, 0.2422±0.019;all P<0.05). The expression bands of PADI4 were observed in Western blot 3 weeks after initial immunization, the thickest in the 4th week, and decreased in the 6th week. The expression bands of PTPN2 were observed at all the time points, with no obvious time-dependent trend.
Conclusions PADI4 and PTPN22 are obviously correlated with CIA in rat model. PADI4 is expressed at early stage of the disease, while the expression of PTPN22 sustains throughout the course.

To clone mouse phage antibodies against H-Y antigen from a phage antibody library, three cycles of affinity enrichment of the mouse phage antibody library with male spleen cells and two cycles of nonspecific absorption with female spleen cells were performed. The presence of mouse Fab on the phage surface was determined by ELISA and sequence analysis. 9 of 15 strains can bind to male spleen cells with the specific activity. Recombination rate of the phage antibody library clones is 60%. Sequence analysis of the PCR products of plasmid DNA of E5 clones show VH and Vkappa had common characteristics shared by other known variable region of antibodies. The mouse phage Fab antibody could be used for identifying H-Y antigen, and for the development of sex determination of early embryos in mammals.
Animals ; Base Sequence ; Cloning, Molecular ; Enzyme-Linked Immunosorbent Assay ; Female ; H-Y Antigen ; analysis ; Immunoglobulin Fab Fragments ; genetics ; immunology ; Isoantibodies ; genetics ; immunology ; Male ; Mice ; Molecular Sequence Data ; Peptide Library

This study is to explore the possible mechanisms of the antidepressant-like effect of agmatine. By using two traditional "behavior despair" model, tail suspension test and forced swimming test, we examined the effects of some monoamine receptor antagonists (including beta-adrenergic receptor antagonist propranolol, beta-adrenergic receptor antagonist/5-HT1A/1B receptor antagonist pindolol, alpha2-adrenergic receptor antagonists yohimbine and idazoxan and 5-HT3 receptor antagonist tropisetron) on the antidepressant-like action of agmatine in mice. Activity of adenylate cyclase (AC) in the synapse membrane from rat frontal cortex was determined by radioimmunoassay. Single dose of agmatine (5-40 mg x kg(-1), ig) dose-dependently decrease the immobility time in tail suspension test in mice, indicating an antidepressant-like effect. The effect of agmatine (40 mg x kg(-1), ig) was antagonized by co-administration of beta-adrenergic receptor antagonist/5-HT1A/1B receptor antagonist pindolol (20 mg x kg(-1), ip), alpha2-adrenergic receptor antagonists yohimbine (5-10 mg x kg(-1), ip) or idazoxan (4 mg x kg(-1), ip), but not beta-adrenergic receptor antagonist propranolol (5-20 mg x kg(-1), ip) and 5-HT3 receptor antagonist tropisetron (5-40 mg x kg(-1), ip). Agmatine (5-40 mg x kg(-1), ig) also dose-dependently decrease the immobility time in forced swimming test in mice. The effect of agmatine (40 mg x kg(-1), ig) was also antagonized by pindolol (20 mg x kg(-1), ip), yohimbine (5-10 mg x kg(-1), ip), or idazoxan (4 mg x kg(-1), ip). Incubation of agmatine (0.1-6.4 micromol x L(-1)) with the synaptic membrane extracted from rat frontal cortex activated the AC in a dose-dependent manner in vitro. While the effect of agmatine (6.4 micromol x L(-1)) was dose-dependently antagonized by pindolol (1 micromol x L(-1)) or yohimbine (0.25-1 micromol x L(-1)). Chronic treatment with agmatine (10 mg x kg(-1), ig, bid, 2 w) or fluoxetine (10 mg x kg(-1), ig, bid, 2 w) increased the basic activity, as well as the Gpp (NH)p (1-100 micromol x L(-1)) stimulated AC activity in rat prefrontal cortex. These results indicate that regulation on 5-HT1A/1B and alpha2 receptors, and activation AC in the frontal cortex is one of the important mechanisms involving in agmatine's antidepressant-like action.
Adenylyl Cyclases ; metabolism ; Adrenergic alpha-Antagonists ; pharmacology ; Adrenergic beta-Antagonists ; pharmacology ; Agmatine ; administration & dosage ; pharmacology ; Animals ; Antidepressive Agents ; administration & dosage ; pharmacology ; Behavior, Animal ; drug effects ; Depression ; metabolism ; physiopathology ; Dose-Response Relationship, Drug ; Fenclonine ; pharmacology ; Idazoxan ; pharmacology ; Male ; Mice ; Pindolol ; pharmacology ; Random Allocation ; Rats ; Rats, Wistar ; Receptors, Biogenic Amine ; antagonists & inhibitors ; Serotonin 5-HT1 Receptor Antagonists ; Swimming ; Synapses ; enzymology ; Yohimbine ; pharmacology

OBJECTIVETo investigate the life style, genetic and occupational risk factors of metabolic syndrome (MS) among policemen.

METHODS1:4 matched case-control study was used, based on physical examination data of Tianjin Policemen in 2010, 708 patients with MS were randomly selected as cases, which were matched with 2832 healthy controls on the basis of sex and age (+/- 1 year). An epidemiological investigations on the past exposure status of several possible risk factors was conducted, and the data were analyzed with conditional logistic regression.

RESULTSFifteen factors related to exposure were identified for MS through univariate conditional logistic regression analysis. Multivariate conditional logistic regression analysis suggested that, seven factors, such as family history of hypertension (OR = 2.406, 95% CI: 1.946-2.975), family history of diabetes (OR = 1.301, 95% CI: 1.043-1.623), smoking (OR = 1.357, 95%CI: 1.010-1.823), snoring (OR = 1.268, 95% CI: 1.043-1.543), work intensity (OR = 4.603, 95% CI: 3.767-5.623), occupational stressful events (OR = 1.524, 95% CI: 1.209-1.922), security policemen (OR = 1.453, 95% CI: 1.127-1.872) and criminal investigation policemen (OR = 2.792, 95% CI: 2.168-3.596), could significantly increase the risk of disease development, but dairy products (OR = 0.782, 95% CI: 0.619-0.989) was a protect factor for MS. The results from population attributable risk factors analysis showed that the control of smoking, snoring, work intensity, occupational stressful events can decreased the risk of MS to 16.26%, 11.71%, 56.87% and 8.97%, respectively.

CONCLUSIONMetabolic syndrome has became a significant public health problem among policemen, it's necessary to take measures on life style, occupational risk factors for reducing the incidence of MS, and improving the health level among policemen.

Adult ; Case-Control Studies ; Factor Analysis, Statistical ; Humans ; Logistic Models ; Male ; Metabolic Syndrome ; epidemiology ; genetics ; psychology ; Middle Aged ; Occupational Health ; Police ; Risk Factors ; Young Adult

BACKGROUNDInterleukin-23 (IL-23) is a pro-inflammatory cytokine that is thought to be central to the development of autoimmune diseases. This study was conducted to determine whether or not the serum concentration of IL-23 is elevated in patients with rheumatoid arthritis (RA), and to determine the relationship between the IL-23 level and disease activity in RA patients.

METHODSSerum samples were obtained from 59 patients with RA and 30 healthy controls. The clinical parameters of disease activity were determined, including the 28-joint disease activity score (DAS28), C-reactive protein (CRP), rheumatoid factor (RF) levels, and the degree of bony erosions based on X-rays. The levels of IL-23 and IL-17 were determined by enzyme-linked immunosorbent assay (ELISA). The correlations between the serum levels of IL-23 and disease activity parameters of patients with RA were determined.

RESULTSThe serum IL-23 level was significantly elevated in patients with RA compared to healthy controls. The serum IL-23 levels in the RA patients correlated with IL-17 and CRP levels, and the DAS28. The levels of IL-23 based on X-ray classification phase I, II, III, and IV were gradually elevated in RA patients.

CONCLUSIONSThe levels of serum IL-23 in RA patients were higher than in healthy controls. Thus, elevated serum IL-23 levels may be useful markers to detect active RA. In addition, IL-23 is involved in disease progression and bony erosions in patients with RA.

Adult ; Aged ; Arthritis, Rheumatoid ; blood ; pathology ; Enzyme-Linked Immunosorbent Assay ; Female ; Humans ; Interleukin-23 ; blood ; Male ; Middle Aged

[ Objective] To discuss the characteristics of death among elderly people aged 60 years and above in Shanghai, who were classified into different age groups, and to provide a basis for making public health policy. [ Methods] On the basis of the data covering whole population death registry system in Shanghai, data on the elders aged 60-plus was collected and classified into 3 age groups according to WHO standards for descriptive analysis. [ Results] The crude death rate among the elders aged 60-plus in shanghai in 2014 was 3 001.76/105 ,accounting for 88.37%of the total mortality.The crude death rate of male was higher than that of female in each age group.The main causes of death in 60-74 age-group were cancer and coronary heart disease ( CHD) which had shorter course of disease with worse prognosis. The main causes of death in 75-89 age-group were cardiovascular and cerebrovascular diseases and chronic obstructive pulmonary disease ( COPD) which had longer course.The main causes of death in 90-pluse group included functional degradation and accidental fall, apart from cardiovascular and cerebrovascular diseases and COPD.And 54.35%of the elders aged 60-plus died in hospital, while 34.12%at home,and 6.63%at nursing home.The proportion of death at home was higher in non-central urban area than in central urban area.And the proportion of death in hospital decreased with increasing age. [ Conclusion] The proportion of the elderly death was large in total mortality.As the characteristics of death varied in different age groups, government should adopt different prevention and control measures.Rational allocation of medical and rehabilitation resources, as well as terminal care, need more attention and exploration by all institutions concerned.

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.

Jiangsu province is rich in intertidal marine resources,especially gastropod and bivalve shellfish.In this study,we performed shotgun proteomic analysis on shellfish from Lüsi,Jiangsu to identify protein content.In total,265 proteins were identified in nine species of shellfish using nano LC-MS/MS.The highest number of proteins were identified in the gastropod Hemifusus tuba (HF) (105 proteins),whereas the bivalve Scapharca subcrenata (SSC) produced the fewest (39 proteins).The most abundant proteins are constituents of cytoskeleton,including actin,tubulin,tropomyosin,myosin heavy chain,as well as histone proteins.Additional proteins were also identified,including ribosomal proteins,enzymes such as arginine kinase and ATP synthase,and housekeeping proteins such as elongation factor and heat shock protein.Proteins were classified based on cellular components and molecular function using the principal component analysis (PCA).Results of PCA analysis revealed that Sinonovacula constrzcta (SC),Bullacta exarata (BE) and Hemifusus tuba (HF) showed similar protein profiles,whileSolen strictus (SS),Meretrix meretrix (MM),Ruditapes philippinarum (RP),and Mactra veneriformis (MV) also showed similar protein profiles.Hierarchical clustering results of heat map studies showed that RP and Mactra antiquate (MA) were classified into one group.SC,BE,HF,MM,SSC,and MV were classified into another group.SS showed the furthest distance from this group.We established a method for analyzing and characterizing the protein composition characteristics of predominant shellfish species in Jiangsu providence's intertidal zone area,which provides basic data for the development of shellfish resources.

Objective To study the epidemical characteristics of influenza/novel influenza A (H1N1) in Shanxi province from 2009 to 2010, and to provide scientific foundations for predicting and controlling the pandemic outbreak of influenza/novel influenza A (H1N1) effectively. Methods All samples were collected from cases that resemble influenza cases in sentinel hospital and influenza outbreak.The influenza were detected by PCR and isolated by MDCK cell culture method. Finally, Shanxi province surveillance data from May, 2009 to April, 2010 of influenza like illness (ILI) cases and pathogen detections were analyzed. Results In Shanxi province, influenza viruses kept activation in whole year. The predominant pandemic strain in 2009 was novel influenza A (H1N1) virus. The strong peak was around November, 2009 [positive rate: 58.1% , novel influenza A( H1N1 ) of the total: 88. 1%]. As well, the people infected influenza caused by novel influenza A (H1N1) were mainly under 59-year-old, and the higher positive rates were concentrated in the people from 5-year-old to 24-year-old. In 2010, influenza B (Victoria) viruses were mainly detected from clinical specimens and became the dominant strain.Conclusion: Surveillance of Influenza liue illnes, (ILI) and etiology, which can promptly reflect the influenza epidemic situation, play a significant role for understanding epidemic rule of influenza/novel influenza A( H1N1 ).