BACKGROUND:Neuronal calcium sensor-1 protein has a variety of different neuronal functions and has a high distribution in different areas of the brain. A single residue R102Q mutation in human neuronal calcium sensor-1 protein is demonstrated to be associated with autism disease. The experiment studies have reported that this R102Q mutant has essential conformation changes in local area of the neuronal calcium sensor-1. OBJECTIVE: To wel understand the specific reasons of the R102Q mutation of the neuronal calcium sensor-1 to the conformational dynamic changes. METHODS:Six independent extensive al-atom molecule dynamic simulations during 0-450 ns were conducted. RESULTS AND CONCLUSION: We have found that (1) there is no obvious recombination during the simulations between wild type and mutant type, but R102Q mutant alters the helix and makes the structure of the protein more stable; (2) R102Q mutation alters the salt bridges, reduces the flexibility of L2, and makes L3 extend in hydrophobic crevice. These results reveal that the helix plays an important role in the structural stability, and salt bridge is the important reason for the dynamic changes of neuronal calcium sensor-1 protein. This study may provide a structural insight into the function of protein deficiency associated with R102Q mutant.

BACKGROUND:Research front and hotspots of neuronal calcium sensor-1 are always the focus for the researchers in this field.
OBJECTIVE:To probe the research front and hotspots of neuronal calcium sensor-1 with the methods of quantitative analysis.
METHODS:The methods of co-cited articles analysis and word frequency analysis were used in the article. The objects were 363 articles from Web of Science by US Institute for Scientific Information (ISI) about the neuronal calcium sensor-1 from 1982 to 2014. The network of co-cited articles and keywords was showed in visualization mapping by using CiteSpace III in which the burst nodes represented the high impact hot papers and the most frequently used keywords, and revealed the research frontier and the hot spots of neuronal calcium sensor-1. RESULTS AND CONCLUSION:The physiological functions of neuronal calcium sensor-1 are the research
frontier and the hot spots. The transformational point in time spot of the hotspots is during 1994 to 1996, 2000, 2008, 2012;and the different research focus showed in each stage:the structure and characterization of the protein during 1992-2000 and the protein function and the role during 2004-2012 are the research hotspots, while during 2008-2014 the hotspots place extra emphasis on the higher function (e.g. memory) and several diseases(such as schizophrenia, cancer, autism, depression, senile dementia, neuron damage, etc). The determination of the research frontier domains and hot spots of neuronal calcium sensor-1 wil indicate the goal and direction for the further studies.

BACKGROUND: Physiological functions, structural fold and unfolding of neuronal calcium sensor-1 (NCS-1) have been explored in a series of experiments, and then the possible mechanism models and key factors for remaining the structural stability are raised. But many functional models cannot be verified due to the limitations of resolution of the time and space and complex protein structure. The experimental phenomena and hypothesis or models may be tested at the atom levels by molecular dynamics, and the new structure may be predicted to provide basis for model establishment and functional mechanisms. OBJECTIVE: To overview the research process of physiological functions and mechanisms of NCS-1 using the experimental method and molecular dynamics simulations, thereby providing basis for future research.METHODS: PubMed database was retrieved for the literatures addressing NCS-1 using the English subject term Neuronal Calcium Sensor-1 or Neuronal Calcium Sensor1 or Neuronal Calcium Sensor 1 or NCS-1. Finally, 72 articles were included in result analysis based on the inclusion and exclusion criteria. RESULTS AND CONCLUSION: The theoretical models of NCS-1 in secretion regulation, dopamine D2 receptor regulation, adenosine A2A receptor regulation in hepatocytes and Ca2+ regulation in myocardial cytoplasm and nuclei with different stimuli are put forward. The key factors to remaining structural stability are analyzed and summarized by modular dynamics simulation in view of structure. It is recommended to combine these two methods in order to deeply understand the protein functional mechanisms, thereby pushing the in-depth study.

Objective To study the effect of alpha-melanocyte stimulating hormone (α-MSH) and its novel analogue ( STY39 ) on the production of tissue factor ( TF ) and tissue factor pathway inhibitor (TFPI) stimulated by lipopolysaccharide (LPS) in primary mouse brain microvascular endothelial cells (MBMECs). Methods Female BALB/c mice were selected,purified and primarily cultured for 5 to 7 days. Immunofluorescence assay was use to detect the Ⅷ factor related antigen and identify the MBMEC model. The MBMECs were divided into eight groups:PBS control group, LPS stimulation group, after LPS stimulation 1,2,and 3 h adding 10 -7 mol/Lα-MSH groups or STY39 group (LPS+α-MSH,LPS+STY39) ( n=4 wholes in each group) . The cell culture supernatant and cells were collected at 6 and 8 h after LPS stimulation. An enzyme-linked immunosorbent assay was used to detect the concentrations of TF and TFPI in cell supernatant. RT-PCR was used to detect the expression levels of TF mRNA. Results (1) LPS could induce MBMEC to produce TF and TFPI proteins. The level of TF in the cell culture supernatant reached the peak at 6 h,and the level of TFPI reached the peak at 8 h. (2) At 1,2,and 3 h after LPS stimulating MBMEC,10 -7mol/L α-MSH or STY39 were given. They could significantly decrease the TF protein content in the cell supernatant (P<0. 01),especially the effects of giving α-MSH or STY39 were most significant at 1 h after LPS stimulation (P<0. 05). The effect of STY39 for decreasing TF content was more significant than that of α-MSH (P<0. 05);however,α-MSH and STY39 did not have significant up-regulating effects for LPS inducing MBMEC to produce TFPI. (3) After LPS stimulation,10 -7 mol/Lα-MSH or STY39 were given at different time points. They significantly down-regulated the expression level of MBMEC TF mRNA (P<0. 01). The effect was most significant at 1 h time point (P<0. 05),but there was no significant difference in the effects betweenα-MSH and STY39. Conclusion Bothα-MSH and STY39 can suppress LPS-induced primary MBMEC to produce TF protein and express TF mRNA,and the effect of administration is better after 1 h LPS stimulation. The suppressive effect of STY39 on the production of TF protein is superior toα-MSH.

Objective To evaluate the effect of α-melanocyte stimulating hormone (α-MSH) and its novel analogue STY39 on the production of tissue factor pathway inhibitor (TFPI) in mice with endotoxemia.Methods Female BALB/c mice were randomly divided into eight groups with 9 mice in each group.Endotoxemia was reproduced by intraperitoneal injection of lipopolysaccharide (LPS,25 μg/kg) and D-galactosamine (D-Gal,100 mg/kg).The animals of the control group were given phosphate buffered solution (PBS) instead.In the experimental groups,the mice were injected intraperitoneally with 2.5 mg/kg α-MSH or STY39 at 1,2 or 3 hours following LPS injection.The orbital blood was collected at different time points,and tissues of lung,liver,and kidney were collected 8 hours after the administration of LPS.The plasma TFPI levels were determined by enzyme linked immunosorbent assay (ELISA),and the expression of TFPI mRNA in different tissues was determined with reverse transcription-polymerase chain reaction (RT-PCR).Results The plasma TFPI levels (μg/L) began to increase (11.84 ± 1.55) in the endotoxemia mice 4 hours after LPS challenge and reached the peak (23.49 ± 1.12) at 8 hours.α-MSH or STY39 treatment at 1,2 or 3 hours after LPS challenge could significantly increase the TFPI content,with the best drug effect at 1 hour after LPS challenge (the blood was collected 8 hours after LPS challenge,α-MSH group:58.79 ± 2.67 vs.28.49 ± 1.69,STY39 group:71.08 ± 2.13 vs.28.49 ± 1.69,both P＜0.01),and the effect of STY39 was better than that of α-MSH (P＜0.01).A small amount of TFPI mRNA expression was observed in each tissue of the healthy mice.After LPS challenge,TFPI mRNA expression was increased in all the tissues,especially in the lung,liver and kidney.α-MSH or STY39treatment at 1 hour after LPS challenge could significantly up-regulate the expression of TFPI mRNA in the lung and liver (A value,α-MSH in lung:51.10 ±2.89 vs.32.43 ±2.51,STY39 in lung:72.11 ±3.48 vs.32.43 ±2.51;α-MSH in liver:43.21 ± 2.12 vs.29.29 ± 2.06,STY39 in liver:66.82 ± 1.76 vs.29.29 ± 2.06,both P＜0.01).The treatment with STY39 at 1 hour after LPS challenge could significantly up-regulate the expression of TFPI mRNA in the kidney (A value:45.21 ± 1.80 vs.30.44 ± 2.23,P＜0.01),but the treatment with α-MSH had no obvious effect (A value:24.61 ± 1.98 vs.30.44 ± 2.23,P＞0.05).The enhancing effect of early administration of STY39 on TTPI mRNA expression in the lung,liver and kidney tissues of endotoxemia mice was more powerful than that of α-MSH (all P＜0.01).Conclusion The early administration of α-MSH or STY39 may up-regulate TFPI production in the mice with endotoxemia,and the effect of STY39 is superior to α-MSH.

As seculars,we often feel unhappy,and it seems all of our unhappiness comes from almost every aspect of life,which is unable to escape.Russell′s view of happiness dug out the root of unhappiness from human nature,and tells us happiness could be obtained by expending one′s interests and necessarily giving away something in life.Russell′s great wisdom opens a door for us to the road of happiness.

Objective:To study the effects of TNF? converting enzyme(TACE)inhibitors on TNF? secretion and develop an approach to interfere inflammation processes.Methods:(1)Stimulate the HL-60 cell lines in vitro with LPS for different time to establish the cellular model of inflammation and simultaneously induce in vivo inflammatoin animal model by LPS.(2)Check the cytotoxic effects of TNF? secretion using MTT colorimetric method for cell proliferaton.(3)Detect the level of expression of TACE carrying out RT-PCR,FCM techniques and immuno-histochemical dying technique.Results:(1)PDQ had the inhibitive effect on TNF?mRNA expression induced by LPS stimulation( P

Objective To investigate the effect of insulin combined with Edaravone therapy on delayed cerebral vasospasm (CVS) after experimental subarachnoid hemorrhage (SAH).Methods 40 New Zealand white rabbits were randomly separated into sham-operated group (n=8),SAH group (n=10),insulin group (n=10) and insulin+Edaravone group(n=12).SAH models were made by injecting blood twice into cisterna magna.30 min later of the first blood injected,the insulin 0.2 U/kg was subcutaneous injected in insulin group and insulin+Edaravone group,3 times daily for 7 d;while,the insulin+Edaravone group received Edaravone 0.5 mg/kg (intravenous injected in ear vein) twice a day for 7 d.7 d after the model made,the basilar artery cross sectional area was measured and the pathological changes were obseved to estimate the degree of CVS.The expression of insulin receptor(InRa) in vascular endothelial cell was detected by immunohistochemical staining.Results The basilar artery cross sectional area between insulin+Edaravone group[(0.46?0.3)mm2]and the sham-operated group[(0.48?0.4)mm2] was no significant difference;but they were obviously bigger than those in SAH group[(0.25?0.3) mm2] and the insulin group [(0.30?0.3)mm2](all P

Objective To explore the clinical value of laparoscopy combined with choledochoscopy in repeat surgery for hepatolithiasis. Methods The clinical data on 86 patients who had undergone repeat surgery for hepatolithiasis during January 2010 to December 2015 were retrospectively analyzed. 36 patients received laparoscopy combined with choledochoscopy(laparoscopy group),while 50 patients received laparotomy(laparotomy group). Surgical duration,intraoperative blood loss,intraoperative transfusion,stone clearance rates,length of postopera-tive hospital stay,and rate of complications were observed and analyzed. Results There were no significant differ-ences in surgical duration,intraoperative blood loss,intraoperative transfusion,stone clearance rates,and rate of complications between the two groups(P>0.05). Length of postoperative hospital stay was significantly shorter in the laparoscopy group than in the laparotomy group(P < 0.05). There was no significant difference in recurrence rates of stone and cholangitis within the follow-up period(P>0.05). Conclusions Use of laparoscopy combined with choledochoscopy in repeat surgery for hepatolithiasis is safe and feasible and has a satisfactory efficacy.

Objective To study the effect of GnRH on the secretion of 5-HT in the stomach. Methods The GnRH-A Alarelin was directly injected into the stomach of rats to observe the change of density of 5-HT immunoreactive positive cell in the stomach and small intestine by immunohistochemical SABC method and 5-HT level in the circulating blood by HPLC-ECD, Results Compared with the control group,which was injected with saline into the stomach,the density of immunoreactive positive cell was significantly increased in the stomach and small intestine in the GnRH-A treatment group;while 5-HT level in the circulating blood was significantly reduced in the experimental group.Conclusion GnRH can inhibit the releasing of 5-HT but not change its synthesis.