Significance of Vesicle-Associated Membrane Protein 8 Expression in Predicting Survival in Breast Cancer / 한국유방암학회지

PURPOSE: Vesicle-associated membrane protein 8 (VAMP8) is a soluble N-ethylmaleimide-sensitive factor receptor protein that participates in autophagy by directly regulating autophagosome membrane fusion and has been reported to be involved in tumor progression. Nevertheless, the expression and prognostic value of VAMP8 in breast cancer (BC) remain unknown. This study aimed to evaluate the clinical significance and biological function of VAMP8 in BC. METHODS: A total of 112 BC samples and 30 normal mammary gland samples were collected. The expression of VAMP8 was assessed in both BC tissues and normal mammary gland tissues via a two-step immunohistochemical detection method. RESULTS: The expression of VAMP8 in BC tissues was significantly higher than that in normal breast tissues. Furthermore, increased VAMP8 expression was significantly correlated with tumor size (p=0.007), lymph node metastasis (p=0.024) and recurrence (p=0.001). Patients with high VAMP8 expression had significantly lower cumulative recurrence-free survival and overall survival (p < 0.001 for both) than patients with low VAMP8 expression. In multivariate logistic regression and Cox regression analyses, lymph node metastasis and VAMP8 expression were independent prognostic factors for BC. CONCLUSION: VAMP8 is significantly upregulated in human BC tissues and can thus be a practical and potentially effective surrogate marker for survival in BC patients.

Identification of differentially expressed proteins and validation of the changes of N-ethylmaleimide-sensitive factor in rats with focal cerebral ischemia after transection of the cervical sympathetic trunk / 华中科技大学学报（医学）（英德文版）

Stellate ganglion blockade (SGB) protects patients from focal cerebral ischemic injury, and transection of the cervical sympathetic trunk (TCST) in a rat model can mimic SGB in humans. The purpose of this study was to investigate the mechanisms underlying the neuroprotective effects of TCST on neuronal damage in the hippocampus in a rat model of middle cerebral artery occlusion (MCAO) in an attempt to elucidate the neuroprotective effects of SGB. The modified method of Zea Longa was used to establish the permanent MCAO model. Male Wistar rats were randomly divided into three groups: sham-operated group, MCAO group, and TCST group. The animals in TCST group were sacrificed 48 h after TCST which was performed after the establishment of the MCAO model. Proteins were extracted from the ipsilateral hippocampus and analyzed by two-dimensional difference gel electrophoresis (2D-DIGE) and peptide mass fingerprinting (PMF). The levels of N-ethylmaleimide-sensitive factor (NSF) were measured as well. The results showed that 11 types of proteins were identified by 2D-DIGE. The expressions of eight proteins were changed both in the sham-operated and TCST groups, and the expressions of the other three proteins were changed in all three groups. Moreover, the expression of NSF was higher in the TCST group than in the MCAO group but lower in the MCAO group than in sham-operated group. The ratio of NSF expression between the MCAO group and shamoperated group was -1.37 (P<0.05), whereas that between the TCST group and MCAO group was 1.35 (P<0.05). Our results imply that TCST increases the expression of NSF in the hippocampus of adult rats with focal cerebral ischemia, which may contribute to the protection of the injured brain. Our study provides a theoretical basis for the therapeutic application of SGB to patients with permanent cerebral ischemia.

Anti-biofilm and anti-adherence activity of Glm-U inhibitors.

Background: Intravascular catheters and urinary catheters are an important source of hospital-acquired infections. Many microorganisms colonize indwelling catheters, including central venous catheters (CVCs) forming biofilms and cause infections that are difficult to treat. Although various methods have been employed to reduce biofilms, enzymes involved in bacterial cell wall synthesis could provide novel targets for the development of anti-biofilm agents. N-Acetylglucosamine-1-phosphate uridyltransferase (GlmU) is an essential enzyme in aminosugars metabolism and catalyzes the formation of uridine-diphospho-N-acetylglucosamine (UDP-GlcNAc), an important precursor in the peptidoglycan and lipopolysaccharide biosynthesis of Gram-positive and Gram-negative bacteria. Previous study has been conducted on the anti-biofilm effect of GlmU inhibitors such as N-ethyl maleimide (NEM) and NEM analogs along with a cationic polypeptide protamine sulfate (PS), which enhanced its anti-biofilm activity. AIM: The present study aimed at finding the effect of sub-inhibitory concentrations of N-ethyl maleimide (NEM) and protamine sulfate (PS) on the biofilms produced by Pseudomonas aeruginosa and Enterococcus spp. isolated from cases of catheter-associated UTI as well as Klebsiella pneumoniae and Staphylococcus aureus isolated from cases of catheter-related bloodstream infections (CRBSI). Materials and Methods: In order to enhance the activity of NEM and to develop a broad-spectrum anti-microbial composition, NEM (50 μg/ml) was combined with protamine sulfate (50 μg/ml) and tested for anti-biofilm activity using a standard quantitative biofilm assay method. Results and Conclusion: It was observed that NEM had no effect on the biofilm produced by Pseudomonas aeruginosa as well as by Enterococcus spp. NEM also caused a significant decrease in biofilm production by Staphylococcus aureus while it had no effect on the biofilm produced by Klebsiella pneumoniae. There was a significant synergistic inhibitory effect on Staphylococcus aureus and Enterococcus spp., whereas Pseudomonas aeruginosa and Klebsiella pneumoniae remained unaffected. Combination of GlmU inhibitor-plus-protamine sulfate failed to significantly reduce bacterial adherence of Pseudomonas aeruginosa and Klebsiella pneumoniae to catheter and cannula pieces, respectively. We found that the GlmU inhibitor was mainly effective in preventing the adherence and biofilm formation by gram-positive organisms. The combination of NEM and protamine sulfate may, therefore, be tried as anti-infective coatings for medical devices such as catheters and cannulas, and thus help in overcoming microbial resistance in the current era of increasing device-associated hospital infections.

Expression of AMPA receptors and related protein in immobilization stressed rats and effect of Xiaoyaosan / 中国应用生理学杂志

<p><b>AIM</b>To observe protein expression changes of alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptor and related regulatory protein in the hippocampus and amygdala in chronic immobilization stressed rat and Xiaoyaosan's regulatory effect.</p><p><b>METHODS</b>Rats were tied 3 h per day to establish immobilization stress condition and treatment with Xiaoyaosan. After 7 days and 21 days stress, the protein expression of AMPA receptor subunit (GluR2/3), N-ethylmaleimide sensitive factor (NSF) and protein interacting with C-kinase 1 (PICK1) in hippocampus and amygdala were detected by using Western blot techniques.</p><p><b>RESULTS</b>The expression of GluR2/3, NSF in dentate gyrus (DG) and amygdala were markedly attenuated (P < 0.05) and PICK1 in CA1 region were significantly increased (P < 0.05) in 7 d immobilization stressed rats while 7 days xiaoyaosan treatment showed an effective regulatory result to PICK1's changes. Under 21 days immobilization stressed condition, the expression of GluR2/3, NSF in CA1 region showed an increasing trend, and GluR2/3 showed a markedly increase (P < 0.01), but showed an significantly decreased trend in amygdala, Xiaoyaosan showed an effective result to such changes above (P < 0.05). The expression of PICK1 showed increasing trend in amygdala and xiaoyaosan could lower its expression (P < 0.05).</p><p><b>CONCLUSION</b>There are different trends of the expression of AMPA receptor in repeat short-term stress versus chronic immobilization stress, and in hippocampal CA1 region versus amygdala. Xiaoyaosan has better regulation effect on the expression of AMPA receptors in the condition of chronic immobilization stress than those of repeat shortterm stress.</p>

Changes of central AMPA receptor subunits and related protein mRNA expression in immobilization stressed rats and effect of Xiaoyaosan on them / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To observe the changes of alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptor subunit and related regulatory protein mRNA expression in the hippocampus and amygdala in immobilization stressed rats and effect of Xiaoyaosan (XYS) on them.</p><p><b>METHODS</b>Immobilization model rats were established by binding for 3 h per day and intervened with XYS, the expression of AMPA receptor subunit (GluR1-4), N-ethylmaleimide sensitive factor (NSF) and protein interacting with C-kinase 1 (PICK1) mRNA expression in model rats' CA1 and CA3 regions of hippocampus, dentate gyrus and amygdala were detected on day 7 and day 21 after modeling.</p><p><b>RESULTS</b>On day 7, expression of GluR1 mRNA was significantly decreased in CA1 region (P < 0.05) and increased in CA3 region and amygdala (all P < 0.05); expression of GluR2 and GluR3 mRNA in amygdala (all P < 0.05) and GluR4 mRNA in CA1 region (P < 0.01) significantly increased, but the expression of NSF and PICK1 mRNA in amygdala only showed an increasing trend. XYS showed effective regulation on GluR4 mRNA in CA1 region (P < 0.01) and GluR1-3 mRNA expression (P < 0.05, P < 0.01) in amygdala. On day 21, the expression of GluR4 mRNA in CA1 region (P < 0.05) and GluR2 mRNA in dentate gyrus (P < 0.05) markedly lowered and expression of GluR1 mRNA in amygdala increased (P < 0.01); XYS significantly regulated the expression of GluR1 and GluR4 mRNA in CA1 region (all P < 0.05).</p><p><b>CONCLUSION</b>Repeated stress in a short time shows effect on expression of AMPA receptor subunit mRNA stronger than chronic stress. The regulation of XYS to AMPA receptor subunit mRNA expression were obvious in hippocampal CA1 region and amygdala.</p>