Objective To investigate the protective effects of lentivirus mediated Bcl-2-associated athanogene 1L (BAG-1L) over-expression on human neuroblastoma cells (SH-SYSY) induced by hypoxia/re-oxygenation,and to study its effect on the phosphoinositide 3 kinase serine/threonine protein kinase (PI3K/AKT) pathway.Methods SH-SYSY cells were cultured in vitro,and the cells at logarithmic phase were collected,and they were divided into recombined lentiviral infection group [infected by lentivirus containing BAG-1L and green fluorescent protein (GFP) gene],vector control group (infected by lentivirus containing GFP without BAG-1L gene) and cell control group (non-infection).Western Blot was used to detect the expression of BAG-1L in target cells after infection for 48 hours.SH-SY5Y cells were subjected to hypoxia for 8 hours and re-oxygenation for 24 hours,then the cell counting kit-8(CCK-8) was used to detect the cell activity,and the apoptosis was detected by flow cytometry after allophycocyanin labeled annexin V/7-amino actinomycin D (Annexin V-APC/7-AAD) staining.Western Blot was used to detect the protein expressions of BAG-1L,heat shock protein 70 (HSP70),AKT and phosphorylated AKT (p-AKT).Results After infection for 48 hours,exogenous BAG-1L protein bands were observed in recombined lentiviral infection group,but not observed in cell control group and vector control group.After hypoxia/re-oxygenation treatment,the cell viability in recombined lentiviral infection group was significantly higher than that in cell control group and vector control group (A value:0.689 ± 0.036 vs.0.425 ± 0.013,0.400 ± 0.012),apoptosis was significantly decreased [apoptosis rate:(26.97 ± 1.82)％ vs.(36.60± 1.45)％,(35.77 ± 3.74)％],the protein levels of BAG-1L,HSP70 and p-AKT were significantly increased [BAG-1L protein (gray value):2.405 ± 0.167 vs.0.529 ± 0.141,0.601 ± 0.099;HSP70protein (gray value):0.997±0.123 vs.0.634±0.091,0.584±0.106;p-AKT protein (gray value):1.234±0.118 vs.0.661 ± 0.210,0.712 ± 0.199,all P ＜ 0.01],but the protein level of AKT was slightly increased (gray value:1.103 ± 0.269vs.0.646 ± 0.188,0.791 ± 0.326) without statistically significant differences (both P ＞ 0.05).There was no significant difference in all parameters between cell control group and vector control group (all P ＞ 0.05).Conclusion Lentivirus mediated BAG-1L gene over-expression can protect nerve cells against hypoxic injury and apoptosis,and the protective effect may be related to the activation increase of pathway on PI3K/AKT.

Background Studies have determined that nucleotide binding oligomerization domain 2 (NOD2) plays a key role in innate immune response.However,whether NOD2 participates in the nature defense of fungal keratitis is unclear.Objective This study was to investigate the expression and significance of NOD2 on cornea in the initial of Aspergillus fumigatus keratitis (AFK) in rats.Methods Seventy-two adult clean Wistar rats were randomized into the normal control group,only corneal epithelial scraped group and AFK model group,and the AFK models were established by incubating Aspergillus fumigatus to cornea after corneal epithelium was scraped.All the operations were performed in the right eyes of rats.Reverse transcription PCR (RT-PCR) was carried out to detect the expression of NOD2 mRNA in corneal epithelium 4,8,16,24 hours after operation.Twenty-four hours after operation,the expression of NOD2 protein in rat corneas was examined by immunochemistry and immnunofluorescence technology.Also,the rat corneas were obtained for regular histopathological examination.The use and care of the animals complied with Institutional Animal Care and Use Committee Guidebook by NIH.Results All the models were made successfully.RT-PCR revealed that a fewer NOD2 mRNA were expressed on cornea in the normal control group,but the expressing levels of NOD2 mRNA were increased in the only corneal epithelial scraped group and AFK model group.Compared with only corneal epithelial scraped group,the elevated values of NOD2 mRNA expression in the AFK model group were statistically significant at 4,8,16 and 24 hours after operation (t =-0.409,-0.439,-0.534,-0.618,all at P=0.000).The histopathological examination displayed that the cornel tissue had intact structure in the normal control group,and partly corneal epithelial deficiency,slight corneal swelling and fewer neutrophil granulocytes were seen in the only corneal epithelial scraped group.However,corneal ulcer,severe corneal edema and a lot of neutrophil granulocytes were exhibited in the AFK model group.Immunochemistry and immnunofluorescence staining evidenced that weaker expression of NOD2 was visualized in the corneal epithelial and endothelial layers,and obviously enhanced staining was seen in the AFK model group.The expressing levels (absorbancy) were 0.045 ± 0.005,0.050 ± 0.005 and 0.092 ± 0.006 in the normal control group,only corneal epithelial scraped group and AFK model group,respectively,showing a significant increase in the AFK model group compared with the only corneal epithelial scraped group (t =0.042,P =0.000).Conclusions Expression of NOD2 is upregulated in the corneas with AFK,suggesting that NOD2 participates the natural defense in the initial of fungal keratitis.NOD2 may play an important role in the process of anti-fungal innate immune response in cornea.

Objective To investigate the effect of intensive lipid-lowering therapy on the imbalance between inflammatory and anti-inflammatory responses in patients with acute coronary syndrome (ACS).Methods Observed serum levels of hs-CRP and IL-IO in 82 patients with ACS, 17 patients with stable angina, and 22 controls. Forty-one patients with ACS were randomized to take either atorvastatin 10 mg/d (standard lipid-lowering therapy) or atorvastatin 40 mg/d (intensive lipid-lowering therapy) for one month in addition to their routine anti-anginal treatment. Serum levels of hs-CRP, blood lipids, IL-10 were investigat-ed.IL-10 was measured by ELISA. Results The level of hs-CRP in patients with ACS [(11.10 ± 14.30)mg/L] was higher than that in patients with stable angina [(2.47 ± 2.72) mg/L]and controls [(2.34 ± 4.22)mg/L] (P all < 0.05 ). The level of IL- 10 was lower in ACS patients [( 3.94 ± 1.91 ) ng/L] compared with those who had stable angina [(6.31 ± 4.26) ng/L] and controls [(7.76 ± 2.82) ng/L], Pan <0.05. The level of hs-CRP in patients with ACS was decreased and IL-10 was increased after one month treatment with atorvastatin (P < 0.05).The effect of atorvastatin 40 mg/d was more effective than that of atorvastatin 10 mg/d.Conclusions Patients with ACS have higher level of hs-CRP and lower level of IL-10 than those with sta-ble angina. This finding suggests that imbalance of inflammatory and anti-inflammatory responses is related with aggravation of atherosclerotic disease. Intensive lipid-lowering therapy is more effective than standard lipid-lowering therapy on ameliorating the imbalance.

OBJECTIVETo investigate the effects of early postnatal exposure to dieldrin on striatum synaptic development in lactation, adolescence and adulthood of mice.

METHODSThe pups were divided into 5 groups randomly. Three groups were exposed to dieldrin (0.01% DMSO solution) at doses of 0.2, 2.0 and 20.0 microg/kg and two control groups were exposed to DMSO or saline by intraperitoneal injection of every other day from postnatal days (PND) 3 to PND13. The striatum were isolated from brain in lactation (PND14), adolescence (PND36) and adulthood (PND98). Western blot assay was used to detect the expression levels of striatal synaptic proteins.

RESULTSThe postnatal exposure to dieldrin could reduce the level of growth associated protein (GAP43) of striatum in lactation in a dose-dependent manner. In adolescence, the level of glial fibrillary acidic protein (GFAP) in striatum increased and the levels of tyrosine hydroxylase (TH), GAP43 and post-synaptic density protein 95 (PSD95) decreased with exposure doses. The level of Synapsin I decreased in adolescence male mice. The changes of expression levels of GFAP, TH and PSD95 proteins lasted to adulthood.

CONCLUSIONEarly postnatal exposure to dieldrin could affect the expression level of GAP43 protein in striatum. The expression levels of TH and PSD95 proteins in striatum decreased in adolescence and adulthood. These results indicated that the early postnatal exposure to dieldrin may persistently interfere in the striatal synaptic development.

Animals ; Animals, Newborn ; Corpus Striatum ; drug effects ; growth & development ; Dieldrin ; toxicity ; Female ; Glial Fibrillary Acidic Protein ; metabolism ; Male ; Mice ; Mice, Inbred ICR ; Nerve Tissue Proteins ; metabolism ; Post-Synaptic Density ; drug effects

Objective To investigate the feasibility of the arterial wall imaging technology of high-resolution magnetic resonance imaging ( HR-MRI) in the risk assessment of intracranial aneurysm rupture. Methods Fifty-four patients with 66 intracranial aneurysms underwent 3. 0 T HR-MRI multiple sequences arterial wall imaging from November 2013 to March 2015 were analyzed retrospectively. Five patients with ruptured aneurysm were used as a control group. The characteristic differences of aneurysm lesions between an unruptured intracranial aneurysm ( UIA) wall enhancement group and a non-enhancement group were compared. The risk factors for rupture were analyzed according to the size,location, and basic clinical characteristics of aneurysm. Results (1) HR-MRI revealed that whether the aneurysm walls enhanced or not,there were no significant differences in the location size,wide-necked aneurysm or not,and ratios of aneurysm height and neck width (all P >0. 05). (2) The enhancement rates of the aneurysm volume <2 group and ≥2 group were 20%(8/40) and 61. 9%(13/21) respectively,the incidence of the ruptured aneurysm asci was higher than that of UIA,and there was significant difference ( all P<0. 05). There were no significant differences in neck width,rate of aneurysm volume,ratios of aneurysm height and neck width,and enhancement rates among the groups. Conclusion The preliminary results of this study have showed that there is a related trend between the HR-MRI aneurysm wall enhancement and the risk of rupture,but a further large sample follow-up study is needed.

Objective To investigate the prevalence of malnutrition and nutritional support and interventions in geriatric inpatients.Methods The elderly patients (aged ≥ 65 years)from the geriatric demonstration ward were consecutively enrolled from July 2010 to January 2012.MiniNutritional Assessment-short form (MNA-SF) was performed after admission,and data of nutritional support were collected.Results A total of 179 patients were enrolled in this study.According to MNA-SF,42 cases (23.5％)were rated as malnutrition,and 55 cases (30.7％) were rated as at risk of malnutrition.Totally,45 patients received nutritional support.50.0％ (21/42) patients with malnutrition,and 29.1％ (16/55) patients at risk of malnutrition received nutritional support.As to the route of nutrition therapy,the ratio of the enteral to parenteral to combination of enteral and parental nutrition was 4.4 ∶ 1.0 ∶ 1.0.Conclusions The incidence of malnutrition is high in the geriatric inpatients,and routine nutritional risk screening and assessment are essential for the elderly patients.Nutritional support and other comprehensive treatment are in great need,and the enteral nutrition is appropriate and preferred.

Background The rapid diagnosis can win more treating opportunities for patients with fungal keratitis.Even though the fungal culture is the gold standard for the diagnosis of fungal keratitis,it is difficult in early diagnosis due to the long duration of cultivation and false-negative rate.Objective This trial was to explore the clinical value in the rapid diagnosis of fungal keratitis by the combination of corneal scraping with laser scanning confocal microscopy.Methods Corneal scraping and laser scanning confocal microscopy were separately performed in 167 eyes of 167 patients with fungal keratitis.All the eyes were examined by the slit lamp,followed by laser scanning confocal microscope,and then the 10％ KOH corneal smear was examined under the optical microscope.Results The positive rate of diagnosis was 75％ (125/167) by corneal scraping,and that by laser scanning confocal microscopy was 91％ (152/167).The positive rate of examining outcome was significantly higher in laser scanning confocal microscopy than that of corneal scraping (x2 =14.88,P =0.00).The positive results were 114 cases and negative results were 4 cases by two methods,with the concordance rate 70.7％ (118/167).The hyphae or spore were seen in 32 cases by laser scanning confocal microscopy in 42 negative cases by corneal scraping,and in 15 negative cases by confocal laser scanning microscopy,11 positive outcomes were offered by corneal scraping.Conclusions The combined application of corneal scraping with confocal laser scanning microscopy can improve and speed up the diagnosis positive rate of fungal keratitis.

OBJECTIVETo investigate the effect of aluminum chloride on dissociated Ca(2+) in hippocampus neuron cells in mice and the relationship to the learning and memory.

METHODSMale ICR mice in the three intoxicated groups were administered with the double distilled water containing AlCl(3) (10, 50, 300 mg.kg(-1).d(-1)) while those in the control group were administered with the double distilled water for 100 days. The methods of behavior toxicology such as Morris swim maze were used for studying the effect of aluminum chloride on the changes of learning and memory in mice. With calcium sensitive fluorescence indicator Fura-2 as the fluorescent probe, the influence of the subchronic exposure to Al on the dissociated Ca(2+) in hippocampus neuron cells was observed.

RESULTSThe dissociated Ca(2+) in hippocampus neuron cells in the middle dosage group and the high dosage group [(412.25 +/- 53.20), (467.37 +/- 32.85) times] was lower than those in the control group [(293.91 +/- 32.21) times] respectively (P < 0.01), and correlated positively with the dose and dissociated Ca(2+) (r = 0.861, P < 0.01). Compared with the control group, the latent period was lengthened (P < 0.05) in the middle dosage and the high dosage group.

CONCLUSIONThe subchronic exposure to AlCl(3) in mice affects the dissociated Ca(2+) in hippocampus neuron cells. The increase of dissociated Ca(2+) in hippocampus neuron cells may be correlated with the disfunction of cognition in the aluminium intoxicated mice.

Aluminum Compounds ; pharmacology ; toxicity ; Animals ; Calcium ; metabolism ; Chlorides ; pharmacology ; toxicity ; Dose-Response Relationship, Drug ; Hippocampus ; cytology ; drug effects ; metabolism ; Learning ; drug effects ; Male ; Memory ; drug effects ; Mice ; Mice, Inbred ICR ; Neurons ; metabolism

Animals ; Biofilms ; Caenorhabditis elegans ; metabolism ; Genes, Bacterial ; Yersinia pestis ; genetics ; metabolism