Objective To investigate the relationship between CYP19 gene polymorphism and Alzheimer disease. Methods Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) was used to analyze the allele frequency distribution at the Mfe Ⅰ site of CYP19 gene in 102 patients with Alzheimer disease and 121 healthy control subjects. Results The frequencies of CYP19 ml and m2 alleles showed significant differences between the patient group and the control group (66.2% vs 81.0% and 33.8% vs 19.0%, respectively, X2=12.696, P<0.05). In patients with Alzheimer disease, the frequencies ofm1/m1, m1/m2, m2/m2 genotypes of CYP19 gene were 44.1%, 44.1%, and 11.8%, respectively, showing significant differences from the frequencies in the control subjects (65.3%, 31.4%, and 3.3%, respectively, X2=12.384, P<0.05). Conclusion CYP19 gene polymorphism at the Mfe Ⅰ site is associated with the genetic susceptibility to Alzheimer disease.

OBJECTIVETo construct a short hairpin (sh)RNA targeting the gene encoding the MDM2 oncoprotein in order to investigate its role in human hepatocellular carcinoma (HCC) and its potential for use as a gene therapy strategy to inhibit HCC growth in vivo.

METHODSSmall interfering (si)RNAs were designed targeting the MDM2 gene (siMDM2-1 and siMDM2-2) and unrelated sequences (negative control) and cloned into the expression plasmid pGCSilencer-U6-neo-GFP. A HCC mouse model was established by subcutaneous inoculation of HepG2 cells (2 x 10(6) in 0.2 ml) into 20 nude mice. The inoculated mice were divided into four equal groups for tumor-localized injections of saline, negative control siRNA plasmid, siMDM2-1 plasmid, and siMDM2-2 plasmid. Tumor growth was observed daily (by caliper measurement) for one month, when mice were sacrificed by cervical dislocation. The tumor mass was resected for analysis of tumor inhibition rate (% = [(average tumor weight of control group - average tumor weight of treatment group) / average tumor weight of control group x 100]) and effects on MDM2 and p53 mRNA and protein expression (by reverse transcription- PCR and western blotting, both normalized to beta-actin). Significance of between-group differences was assessed by one-way ANOVA or LSD test; pairwise comparisons were made by the Chi-squared test.

RESULTSsiMDM2-1 and siMDM2-2 suppressed the xenografted tumor growth remarkably (60.6% and 54.6% inhibition rates, respectively), significantly reduced the expression ofMDM2 gene (62.8% and 61.6%) and protein (60.7% and 59.5%), and significantly increased p53 gene (47.1% and 45.6%) and protein (45.9% and 44.3%) (all, P < 0.05).

CONCLUSIONshRNA-mediated silencing of the MDM2 gene effectively inhibits HCC tumorigenesis of subcutaneously xenografted HepG2 cells in nude mice, and the mechanism may involve p53.

Animals ; Carcinoma, Hepatocellular ; genetics ; pathology ; Cell Proliferation ; Hep G2 Cells ; Humans ; Liver Neoplasms ; genetics ; pathology ; Male ; Mice ; Mice, Nude ; Plasmids ; Proto-Oncogene Proteins c-mdm2 ; genetics ; metabolism ; RNA Interference ; RNA, Messenger ; genetics ; RNA, Small Interfering ; Transfection ; Tumor Suppressor Protein p53 ; metabolism ; Xenograft Model Antitumor Assays

BACKGROUNDHaptoglobin (Hp) is one of the acute-phase proteins. Recent studies have demonstrated that Hp exerts immunoregulatory and anti-inflammatory actions and may be one of the inhibitory factors of immune reactions in the skin. In this study we investigated the regulation of Hp expression in a human keratinocyte cell line HaCaT by various cytokines and glucocorticoid.

METHODSHaCaT cells were cultured with IL-6 (50 ng/ml), TNF-alpha (20 ng/ml), IFN-gamma (20 ng/ml) or IL-4 (20 ng/ml) with or without 1 micromol/L dexamethasone in 6-well plates for 12, 24 and 48 hours. Both the cells and the supernatants were collected to detect the changes of Hp expression by reverse-transcription PCR, ELISA and immunohistochemistry.

RESULTSThe results showed that Hp expression were elevated at both the mRNA and protein level by the combination of IL-6, TNF-alpha or IL-4 with dexamethasone, whereas the three cytokines alone did not upregulate the Hp expression. IFN-gamma showed no effect on the Hp expression in HaCaT cells.

CONCLUSIONSThese findings suggest that different inflammatory cytokines as well as glucocorticoid may be involved in the regulation of Hp expression in keratinocytes, and this may be one of the negative feedback mechanisms in inflammatory skin diseases.

Cell Line ; Dexamethasone ; pharmacology ; Glucocorticoids ; pharmacology ; Haptoglobins ; analysis ; Humans ; Interferon-gamma ; pharmacology ; Interleukin-4 ; pharmacology ; Interleukin-6 ; pharmacology ; Keratinocytes ; chemistry ; drug effects ; Tumor Necrosis Factor-alpha ; pharmacology

OBJECTIVECigarette smoke extract (CSE) can induce injuries of pulmonary II epithelial cells, activate nuclear factor-kappaB and increase tumor necrosis factor-alpha(TNF-alpha) secretion. This study aimed to investigate whether azithromycin can protect pulmonary II epithelial cells from injuries induced by CSE and relevant mechanisms.

METHODSPulmonary II epithelial cells (A549 cells) were cultured in vitro. After 48 hrs of culture the cells were randomly treated with serum-free DMEM only (blank control group), azithromycin + serum-free DMEM, CSE+ serum-free DMEM or CSE+azithromycin. Eight hours later the morphology of A549 cells, the activity of NF-kappaB and the levels of TNF-alpha were measured by inverted microscope, immunohistochemistry and ELISA.

RESULTSThe morphology and structure of A549 cells were changed, NF-kappaB activity increased (dark brown staining ) and TNF-alpha levels (0.307 +/- 0.036 pg/mL vs 0.234 +/- 0.028 pg/mL)increased in the CSE+ serum-free DMEM group compared with the blank control group (P < 0.01). CSE together with azithromycin treatment recovered partly the morphological injuries of A549 cells. It also attenuated NF-kappaB staining and decreased TNF-alpha levels from 0.307 +/- 0.036 pg/mL (CSE+serum-free DMEM group) to 0.269 +/- 0.009 pg/mL (P < 0.05).

CONCLUSIONSAzithromycin may inhibit NF-kappaB activity, decrease TNF-alpha secretion and thus lessen cytotoxicity of CSE to A549 cells.

Anti-Bacterial Agents ; pharmacology ; Azithromycin ; pharmacology ; Cells, Cultured ; Epithelial Cells ; drug effects ; Humans ; Immunohistochemistry ; Lung ; drug effects ; metabolism ; pathology ; NF-kappa B ; analysis ; Smoke ; adverse effects ; Tobacco ; adverse effects ; Tumor Necrosis Factor-alpha ; analysis

Objective To study the changing trend of prevalence rates of healthcare-associated infection (HAI) and antimicrobial use in hospitalized patients in medical institutions of Hubei Province,and provide a scientific basis for improving HAI management.Methods The cross-sectional survey results of HAI in Hubei Province in 2010, 2012,and 2014 were analyzed.Results The prevalence rates of HAI in 2010,2012,and 2014 were 3.48%(1526/43909),3.03%(1919/63320),and 2.86%(2174/76145)respectively,which showed a downward trend,differ-ence was statistically significant(χ2 =36.44,P <0.01).Antimicrobial usage rate decreased from 54.29% (23838/43909)in 2010 to 41.02% (31238/76145)in 2014,difference was statistically significant(χ2 =2194.09,P <0.01).Among patients receiving therapeutic use of antimicrobial agents,the specimen detection rate increased from 30.49% (4297/14091)in 2010 to 52.13% (10556/20248)in 2014 (χ2 =1593.98,P <0.01).Conclusion The prevalence rate of HAI showed a downward trend in Hubei Province,cross-sectional survey on antimicrobial use showed a gradual decrease.

Medium- and long-chain triglyceride (MCT/LCT) propofol is widely used as an intravenous anesthetic, especially in the intensive care unit. The present study aimed to assess whether MCT/LCT propofol is safe in the hyperlipidemic population for long-term use. Free fatty acids (FFAs) were used to establish high-fat stimulation of HepG2 and Huh7 cells. Subsequently, these cells were treated with propofol at the concentration of 0, 4, or 8 µg/ml for 24 and 48 h. The results indicated that the cell viability was notably decreased when the cells were stimulated with 2 mmol/L FFAs and treated with 12 µg/ml MCT/LCT propofol. Accordingly, we chose 2 mmol/L FFAs along with 4 and 8 µg/ml MCT/LCT propofol for the subsequent experiments. Four and 8 µg/ml MCT/LCT propofol inhibited FFA-induced lipid accumulation in the cells and significantly reversed acetyl coenzyme A carboxylase (ACC) activity. In addition, MCT/LCT propofol not only significantly promoted the phosphorylation of AMPK and ACC, but also reversed the FFA-induced decreased phosphorylation of AMPK and ACC. In conclusion, MCT/LCT propofol reverses the negative effects caused by FFAs in HepG2 and Huh7 cells, indicating that MCT/LCT propofol might positively regulate lipid metabolism.
Acetyl-CoA Carboxylase ; AMP-Activated Protein Kinases ; Cell Survival ; Fatty Acids, Nonesterified ; Hepatocytes ; Intensive Care Units ; Lipid Metabolism ; Liver ; Metabolism ; Phosphorylation ; Propofol ; Triglycerides

Objective To study the effects of human uriilary kallikrein(HUK)on the number of apoptotic cells and the expressions of Bcl-2 and Bax proteins in rats after focal cerebral ischemia and reperfusion(FCIR) injury. Methods Eighty-four Spmque-Dawley(SD)male rats were randomly divided into sham-operated group(n=12),ischemia-reperfusion group(n=36),and HUK-treated group (n=36). Transient focal cerebml ischemia models were established by middle cerebml artery occlusion.Six rats were chosen from sham-operated group,ischemia-reperfusion group,and HUK-treated group for measuring infarct sizes.The rest were used to evaluate neurologic fhnction impaiment and measure the nunlber of apoptotic cells and Bcl-2 or BaX protein positive cells in cerebral cortex with TUNEL and immunohistochemistry.The latter 2 groups were subdivided into 6,12,24,72,168 h reperfusio groups (each n=6). Results The neurologic function impairmlent score,the infarct sizes,the apoptotic cells and the expression of Bax protein of HUK-treated group at different time points (except 168 h group)significantly decreased compared wilh those of ischemia-reperfsion group (p＜0.05).The expression of Bcl-2 protein of HUK-treated group at different time points(except 168 h group) remarkably increased compared with that of ischemia-reperfusion group(P＜0.05). Conclusions HUK can excrt a protection against FCIR injury, maybe through up-regulating Bcl-2 and down-regulating Bax protein in the initial 3 d of FCIR injury to decrease the number of apoptotic cells

OBJECTIVETo investigate the possible mechanism of local hyperthermia in the treatment of warts through detecting the differences in CD1a/CD83 of Langerhans cells (LCs) in émigrés from HPV-infected skin, as compared to normal skin.

METHODSConfocal microscopy were performed on Condyloma Accuminatum (CA)and normal skin; Freshly taken biopsies of CA and normal skin were subjected to surface heating at 37 degrees C, 42 degrees C and 45 degrees C respectively, for 30 mins. Flow cytometry was used to determine the CD1a/ CD83 changes of LCs in émigrés from CA and normal skin.

RESULTSBy confocal microscopic observation, there were practically no CD1a+ LCs that expressed CD83 in the epidermis of both normal skin and CA. The proportions of CD1a+/CD83 LCs were significantly increased with increased temperatures in émigrés from both normal skin and CA. At each given temperature, the numbers of LCs in émigrés from CA were greater than those from normal skin.

CONCLUSIONLocal hyperthermia can promote migration and maturation of LCs in HPV-infected skin and accordingly stimulate the immune system to treat warts.

Adult ; Cell Movement ; immunology ; Female ; Humans ; Hyperthermia, Induced ; In Vitro Techniques ; Langerhans Cells ; cytology ; immunology ; Papillomaviridae ; immunology ; pathogenicity ; Skin ; immunology ; virology ; Young Adult

BACKGROUNDThe sensitization and elicitation phases are involved in the immunopathogenesis of contact hypersensitivity (CHS). Langerhans cells (LCs) are believed to play pivotal roles in the sensitization stage of CHS. Local hyperthermia on skin induces the migration as well as maturation of epidermal LCs. Although fever-range whole body hyperthermia and local hyperthermia at 43°C prior to sensitization were reported to suppress CHS, the effects of different temperatures and the timing sequence of local hyperthermia on CHS have not been tackled.

METHODSLocal hyperthermia was applied to murine dorsal skin 3 days prior to, concurrent with, or 2 days post sensitization with fluorescein isothiocyanate (FITC) in BALB/c mice. Local hyperthermia temperatures at 37°C, 39°C, 41°C and 43°C were applied to mouse dorsal skin and the severity of CHS was calculated by measuring the swelling response of the challenged ears.

RESULTSLocal hyperthermia at 39°C, 41°C and 43°C prior to sensitization reduced the severity of CHS, as compared with that at 37°C. The suppression of CHS was temperature dependent in that higher temperature had a stronger effect. On the contrary, the hyperthermia treatments, either concurrent with or post-sensitization, resulted in an enhanced temperature-dependent ear swelling response.

CONCLUSIONSThe severity of murine CHS could be influenced by local hyperthermia at the sensitization stage in a temperature dependent manner. The temporal effect of local hyperthermia suggested a novel factor in interpreting the severity of allergic contact dermatitis.

Animals ; Dermatitis, Contact ; therapy ; Female ; Hyperthermia, Induced ; Langerhans Cells ; physiology ; Mice ; Mice, Inbred BALB C

BACKGROUNDAtopic dermatitis (AD) is characterized by defective skin barrier and imbalance in T helper 1/T helper 2 (Th1/Th2) cytokine expression. Filaggrin (FLG) is the key protein to maintaining skin barrier function. Recent studies indicated that Th1/Th2 cytokines influence FLG expression in keratinocytes. However, the role of Th1/Th2 cytokines on FLG processing is not substantially documented. Our aim was to investigate the impact of Th1/Th2 cytokines on FLG processing.

METHODSHaCaT cells and normal human keratinocytes were cultured in low and high calcium media and stimulated by either interleukin (IL)-4, 13 or interferon-γ (IFN-γ). FLG, its major processing proteases and key protease inhibitor lymphoepithelial Kazal-type-related inhibitor (LEKTI) were measured by both real-time quantitative polymerase chain reaction and Western blotting. Their expression was also evaluated in acute and chronic AD lesions by immunohistochemistry.

RESULTSIL-4/13 significantly reduced, while IFN-γ significantly up-regulated FLG expression. IL-4/13 significantly increased, whereas IFN-γ significantly decreased the expression of kallikreins 5 and 7, matriptase and channel-activating serine protease 1. On the contrary, IL-4/13 significantly decreased, while IFN-γ increased the expression of LEKTI and caspase-14. Similar trends were observed in AD lesions.

CONCLUSIONSOur results suggested that Th1/Th2 cytokines differentially regulated the expression of major FLG processing enzymes. The imbalance between Th1 and Th2 polarized immune response seems to extend to FLG homeostasis, through the network of FLG processing enzymes.

Caspase 14 ; metabolism ; Cell Line, Tumor ; Cells, Cultured ; Dermatitis, Atopic ; metabolism ; Humans ; Immunohistochemistry ; Interferon-gamma ; metabolism ; Interleukin-13 ; metabolism ; Interleukin-4 ; metabolism ; Intermediate Filament Proteins ; metabolism ; Keratinocytes ; enzymology ; metabolism ; Proteinase Inhibitory Proteins, Secretory ; metabolism ; Serine Peptidase Inhibitor Kazal-Type 5 ; Th1 Cells ; metabolism ; Th2 Cells ; metabolism