Abstract:Aquaporin-4（AQP-4） is the most abundant aquaporin in central nervous system（CNS）. Water movement cross cell mem－brane is mainly conducted by AQP-4. Previous studies of AQP-4 mainly focused on edema formation after various brain injuries，but recently AQP-4 has been found to connect with many other brain functions and diseases，such as regeneration and inflammation. Therefore，these researches were reviewed to renew current views on AQP-4 in CNS，hoping to provide new threads for the exploration of drugs against AQP-4.

Abstract:Objective：To investigate the effects of short-term estrogen replacement on myelinated nerve fibers and myelin sheaths in CA1 region of ovariectomized rats. Methods：Female SD rats（10-11 month-old） were bilaterally ovariectomized. One month after o－variectomy，successful ovariectomized rats were randomly divided into placebo group（OVX+O） which received sesame oil for 4 weeks and estrogen replacement group（OVX+E） which received continuous 17β-estradiol replacement treatment for 4 weeks. After 4 weeks，spatial learning capacities of two groups were tested using the Morris water maze. Then，the serum estradiol levels were detected by chemiluminescence method. The myelinated nerve fibers and myelin sheaths in CA1 region were quantitatively investigated by transmission electron microscope and stereological methods. Results：The serum estradiol levels were significantly increased after es－trogen replacement treatment（t=5.688，P=0.000）. The spatial learning capacity of OVX+E rats was significantly improved when com－pared with that of OVX+O rats（F=9.645，P=0.006）. There was no significant difference in the CA1 volume，the volume of myelinated nerve fibers，the volume of myelin sheaths，the mean diameter of the myelinated fibers and the mean thickness of myelin sheaths be－tween two groups（P=0.181，P=0.294，P=0.521，P=0.545，P=0.081，respectively）. The mean thickness of myelin sheaths of the myeli－nated fibers with diameters between 0.6 to 0.7 μm was significantly higher by 14.1% in OVX+E group when compared with that of OVX+O group（t=2.428，P=0.038）. The difference between the mean diameter of myelinated nerve fibers and the mean diameter of axons with the mean diameter of 0.8 to 0.9 μm in OVX+E group was significantly increased by 18.2% when compared with that of OVX+O group（t=2.432，P=0.038）. Conclusions：Short-term estrogen replacement can enhance the spatial learning capacity and can exert pro－tective effects on the myelin sheaths in the CA1 region of middle-aged ovariectomized rats.

Abstract:Objective：To explore the role of epidermal neural crest stem cells（EPI-NCSCs） in the treatment of spinal cord injury（SCI） of rats and changes in expression of brain-derived neurotrophical factor（BDNF） and to investigate the mechanism of EPI-NCSCs re－pairing SCI from the perspective of neurotrophic factors. Methods：EPI-NCSCs were isolated from green fluorescent protein（GFP） transgenic rats for transplantation. Rat models of SCI were made by NYU-Ⅱ impactor（10 g 25 mm injuries force） at T10 level. Then 60 SD rats were randomly divided into blank injury group（group A），medium transplantation group（group B），epidermal neural crest stem cell transplantation group（group C）. EPI-NCSCs were transplanted into the injured region in group C at 1 week after establishing SCI model. Group B used the DMEM/F12 to take place of EPI-NCSCs. Nothing was done in blank injury group. Then locomotor function score was appraised at every week after the transplantation. Expression of BDNF in three groups was detected by ELISA，immunohistochemistry，immunofluoresence and Western blot respectively at 1，3，6 weeks after the transplantation. Results：After 2 weeks of transplantation，locomotor function score was higher in group C than in group A（2，3，4，5，6 weeks：P=0.003，0.000，0.000，0.000，0.000） and group B（2，3，4，5，6 weeks：P=0.440，0.000，0.006，0.000，0.008）. Based on the results of ELISA，expression of BDNF was higher in group C than in group A（P=0.009） and group B（P=0.013） at 3 weeks after the transplantation；expression of BDNF was higher in group C than in group A（P=0.003） and group B（P=0.003） at 6 weeks after the transplantation. Expression of BDNF was higher in group C than in group A and group B at 3 weeks and 6 weeks after the transplan－tation based on the results of immunohistochemistry and immunofluoresence. Expression of BDNF was higher in group C than in group A（P=0.000） and group B（P=0.000） at 6 weeks after the transplantation based on the results of Western blot. Conclusions：EPI-NC－SCs transplantation can repair SCI by increasing expression of BDNF to improve local microenvironment.

Abstract:Objective：To investigate the effects of the short-term estrogen replacement therapy（ERT） on the white matter and the myelin sheath ultrastructure of the myelinated fibers in the white matter of ovariectomized middle-aged rats in order to provide a sci－entific basis for delaying the process of brain aging and combating demyelination diseases with estrogen replacement therapy in the future. Methods：Middle-aged female sprague-dawley rats of 10-12 month（40 rats） were bilaterally ovariectomized and the serum e－strogen levels were examined one month after the surgery. The female Sprague-Dawley rats（19 rats） with successful castration were ran－domly divided into experimental group（n=10） and control group（n=9）. Experimental rats were subcutaneously injected 17β-estradiol and control rats were subcutaneously injected placebo for 4 weeks. Then，the spatial reference memory capacities of two group rats were tested with Morris water maze. The white matter and the myeli－nated nerve fibers in the white matter of two group rats were quantitatively investigated with the transmission electron mi－croscope technique and the stereological techniques. Results：The spatial reference memory capacity of the experimental rats was significantly enhanced compared with that of the control rats（F=9.645，P=0.006）. The total volume of the myelin sheaths and the difference between outer perimeter and inner perimeter of myelin sheaths in the myelinated fibers of the white matter in experimental rats were significantly increased compared with those of control rats（（32.94±5.97） mm3，（25.93±2.83） mm3，t=2.391，P=0.040；（1.16±0.10） ?滋m，（1.03±0.09） ?滋m，t=2.325，P=0.045）. The correlation analyses suggested that the total myelin sheath vol－ume of the myelinated fibers in the white matter was significantly and negatively correlated with the escape latency on the first day of Morris water maze（r=-0.744，P=0.009）. Conclusions：Estrogen replacement had positive protective effects on the spatial reference memory capacity and the myelin sheaths of the myelinated fibers in the white matter of the middle-aged ovariectomized rats.

Abstract:Objective：To explore the integrated assessment model for stroke risk among Asian population based on the results of Meta analysis. Methods：Based on the data in the Ｃohort study and case-control study of stroke，Meta analysis was used to combine risk factors. Integrated assessment model by logistic regression was built based on the pooled odds ratio（OR）. Results：Seven Cohort stud－ies and 21 case-control studies were included in this study including 17 risk factors. After screening，10 factors were admitted in the model，including total cholesterol≥6.00 mmol/L，systolic blood pressure≥140 mmHg，hypertension，diabetes，coronary heart disease，family history of stroke，smoking，long-term drinking，long-term exercises and high salt diet and the pooled ORs were 1.99，5.09，3.97，1.82，2.10，2.64，1.69，1.37，0.60 and 1.95. Conclusions：Integrated assessment model of stroke among Asian population is built through the systematic review. This model，based on data of evidence-based medicine，successfully avoids the bias of small sample and is of practical significance in the prevention of stroke among Asian population.

Abstract:Objective：To examine the expression of vascular endothelial growth factor（VEGF） and glial fibrillary acidic protein（GFAP） in the retina of mice with chronic brain ischemia as well as to analyze the relationship between VEGF and GFAP under retinal is－chemic injury. Methods：C57 mice were used to establish chronic brain ischemia model. Immunohistochemical staining was performed to observe the distribution and expression of VEGF and GFAP in the mice retina at different ischemic time periods. Results：VEGF and GFAP were unexpressed or weakly expressed in sham operated group. In the operated group，expression of VEGF and GFAP was in－creased at 120 d after ischemia and was significantly increased in intensity and scope at 180 d after ischemia. Conclusions：Expression of VEGF and GFAP are increased in the retina of mice with chronic brain ischemia，which indicates VEGF and GFAP may be involved in the pathogenesis of retinal ischemic injury process.

Abstract:Objective：To detect the expression of vimentin（VIM）in myocardium and to explore the characteristics of myocardial injuries caused by epilepsy. Methods：Models of epileptic rats were established by intraperitoneal injection of pentylenetetrazol. Expression of VIM in myocardium of epileptic rats was examined by pathologic，immunohistochemistry，and Western blot methods. Results：Hemorrhagic and hypoxic ischemic changes were showed in myocardiums of epileptic groups. With the extended duration of epilepsy，and the increases of frequency and time of seizures，there was a trend of aggravation of myocardial necrosis，fibroblast proliferation，as well as a remarkable up-regulation of VIM. Conclusion：Epilepsy can cause myocardial injury and promote the expression of VIM in myocardium.

Abstract:Objective：To explore the expression rule of repulsive guidance molecule B（RGMB） after spinal injury in rats and its cor－relation with the spinal cord functional recovery. Methods：Totally 65 adult male SD rats were divided into A，B，C groups randomly. Chondroitin sulfate（chondroitinase ABC，ChABC） enzyme was not applied to do degradation in group A（n=25）. ChABC was used to do degradation in group B（n=20）. Only spinal cord was exposed in group C（n=20）. Body movements were recorded in three groups at the same postoperative time points. Rats were executed to do specimen detection. Basso-Beattie-Bresnahan（BBB） score was used to evaluate the feeling and movement of rat’s hindlegs. HE staining was employed to calculate the total damage. Real-time PCR was used to detecte RGMB mRNA expression.Immunofluorescence staining was used to observe the double standard expression of glial fibrillary acidic protein（GFAP） and neurofilament-200（NF-200）. Results：Within the same period at 2-4 weeks after the injury，BBB score was obviously higher in group B than in group A（P＜0.05）. Two weeks after the injury，HE staining demonstrated hollow formation in A and B groups；damaged area was significantly smaller in group B than in group A（P＜0.05）. At 2-4 weeks after the injury，GFAP/NF-200 double immunofluorescence staining demonstrated that GFAP expression was gradually lower in group B and immunofluorescence intensity value was significantly higher in group A than in group B（P＜0.05）；NF-200 positive staining fibers passed through the damaged area and immunofluorescence intensity value was significantly higher in group B than in group A（P＜0.05）. RGMB mRNA expression was decreased in group B than in group A（P＜0.05）. Conclusions：After spinal cord injury in rats，RGMB concentration in the microenvironment is negatively correlated with the spinal cord functional recovery. RGMB in－hibitors may be helpful for clinical treatment of spinal cord in－jury.

Abstract:Objective：To observe the therapeutic effect of safflower injection on young patients with mild acute cerebral infarction and the effect of safflower injection on serum interleukin（IL）-6 and IL-10 level of these subjects. Methods：Totally 52 patients with mild acute cerebral infarction，aged 35-40 year-old with 10-15 neurological severity score（NSS） were randomly divided into control group（conventional treatment，27 cases） and treatment group（conventional treatment combined with safflower injection 20 ml per day，25 cases）. Morning blood samples of all patients were monitored at three time points（before the treatment，on the 7th d and 14th d during the treatment） and were collected for IL-6 and IL-10 detection using a double antibody sandwich enzyme-linked immunosor－bent assay. Blood samples of healthy people aged 35-40 years were also obtained as controls. Results：NSS of control group and treatment group on the 7th d and 14th d were all significantly decreased compared with those before treatment（all P=0.000）. On the 14th d，NSS of treatment group was lower than that of control group（P=0.040）. Comparing with that before treatment，serum IL-6 level of control group was significantly decreased on the 14th d（P=0.000） but unchanged on the 7th d（P=0.078）. Serum IL-6 levels of treatment group were markedly decreased on the 7th d and 14th d compared with those before treatment（all P=0.000） and control group respectively（P=0.042，P=0.000）. Serum IL-10 levels of control group and treatment group were significantly increased after treating for 7 d and 14 d compared with those before treatment（P=0.000）. On the 7th d and 14th d，serum IL-10 level was higher in treatment group than in control group（P=0.001，P=0.038）. Positive correlation was observed between IL-6/IL-10 value and NSS in treatment group（r=0.997，P=0.048），but not in control group（r=0.962，P=0.177）. Conclusions：Early application of safflower injection is benefit for young patients with mild acute cerebral infarction. The mechanism might be associated with lowing IL-6/IL-10 value through decreasing IL-6 level and increasing IL-10 level.

Abstract:Objective：To study the resting-state brain activity in patients with minimal hepatic encephalopathy（MHE）. Methods： Sev－enteen patients with MHE and seventeen healthy control（HC） subjects underwent resting-state blood oxygen level dependence functional magnetic resonance imaging and the MRI data were analyzed with the regional homogeneity（ReHo） method. Results：Compared with HC group，MHE group showed lower ReHo in right medial frontal gyrus，bilateral anterior cingulate cortex/medial superior frontal gyrus，right inferior frontal gyrus，left rectal gyrus/orbital gyrus，right thalamus and right cerebellum posterior lobe，while higher ReHo in left insula，right middle frontal gyrus/inferior frontal gyrus and right fusiform gyrus/hippocampus/parahippocampa gyrus（P<0.005，cluster-level corrected）. Conclusion：Patients with MHE demonstrate abnormal neuron activities in multiple brain regions which are related with multiple cognitive control loops.

Abstract:Objective：To observe the effects of Maixuekang capsule on therapeutic efficacy and blood coagulation function in patients with diabetes mellitus complicated with transient cerebral ischemic attack（TIA）. Methods：Totally 80 patients with diabetes mellitus complicated with transient cerebral ischemic attack were divided into 2 groups randomly：40 patients in control group were given enteric-coated aspirin，40 patients in Maixuekang group were given enteric-coated aspirin + Maixuekang capsule. Patients received continuous treatment for three months. TIA attack symptoms，time，number of times within 6 months of treatment were recorded； Intimal medial thickness（IMT），prothrombin time（PT），activated partial thromboplastin time（APTT），thrombin time（TT），fibrinogen（Fib），platelet aggregation rate（PAR），high sensitive C-reactive protein（hs-CRP） at 6 months after the treatment were detected. Results：At 6 months after the treatment，total efficiency of Maixuekang group was significantly higher than that of the control group（ χ2=4.266 6，P＜0.05）. Incidence of cerebral infarction in Maixuekang group was significantly lower than that in the control group（ χ2=4.504 5，P＜0.05）. PT，APTT，TT of patients in Maixuekang group were significantly prolonged compared with those before the treatment and those in control group（P＜0.05）. Fib，PAR and hs-CRP were decreased significantly in Maixuekang group compared with those before the treatment and those in control group（P＜0.01）. There was no significant difference in the incidence of adverse reactions between two groups（ χ2=0.138 8，P ＞0.05）. Conclusions：Maixuekang capsule combined with aspirin is a safe and effective in the treatment of diabetes mellitus complicated with TIA，of which anticoagulation，anti-platelet aggregation and anti-inflammation may be one of the mechanisms.

Abstract:Objective：To explore the protection of edaravone on acute cerebral vasospasm（CVS） after subarachnoid hemorrhage（SAH） in rats and effects of edaravone on expressions of serum S100B and endothelin-1（ET-1）. Methods：Totally 60 Wistar rats were randomly assigned to four groups：control group（group C），normal saline group（group N），simple group of SAH（group SAH） and group of SAH+ edaravone（group SAH+E）. CVS model after SAH was made by injecting rats’ own blood into cisterna twice. Microscopic im－age analysis was used to measure the basal artery diameter and ELISA was employed to determine S100B and ET-1serum levels pre－operatively in group C and at 6，24，72 h postoperatively in others groups. Results：Postoperative basilar artery diameters at the same time were in the following order（from small to large）：group SAH，group SAH + E，group N and group C. There was no statistically significant difference between groups N and C and the differences were statistically significant between the other groups. Factorial anal－ysis suggested that different treatments exerted different effects on the results. Basilar artery spasms sustained for 72 h. At the same time after operation，serum S100B and ET-1 levels were in the following order（from high to low）：group SAH，group SAH+E，group N and group C. There were statistically significant differences between groups but there was no difference between groups N and C. Fac－torial analysis suggested that different treatments exerted different effects on the results. Serum S100B and ET-1 increased consis－tently within 72 h，with statistically significant differences（P<0.05）. Conclusions：Serum S100B and ET-1 levels related with acute CVS after SAH. Edaravone can make serum S100B and an ET-1 level back to normal and thus plays a controlling role in improving acute CVS after SAH.

Abstract:Objective：To analyze the distribution and drug susceptibility of bacteria isolated from cerebrospinal fluid（CSF） in patients who developed nosocomial meningitis following neurosurgical operation and to provide evidences for clinical treatment of meningitis.Methods：Clinical data of pathogens and their drug susceptibility to antibiotics from CSF were collected from January 2008 to Decem－ber 2012. Distribution and drug susceptibility of bacteria were analyzed retrospectively. Results：Positive rate of pathogens was 22.35（114/510） from 510 CSF specimens. There were 114 strains of pathogens including 58（50.88%） G+ bacteria and 56（49.12%） G- bacteria. The most frequently isolated bacteria was coagulase-negative staphylococci（28.95%），followed by acinetobacter baumannii（16.67%），enterococcus spp.（9.65%），pseudomonas aeruginosa（9.65%），klebsiella pneumoniae（6.14%），staphylococcus aureus. The active compounds against G+ bacteria were vancomycin（100%），linezolid（95.92%），piperacillin-tazobactam（88.89%）. The active compounds against G- bacteria were imipenem（66.04%），followed by piperacillin-tazobactam（59.46%），amikacin（55.10%）. Super-infection in bacterial meningitis was observed in 8 patients. Conclusions：G+ bacteria are more common in neurosurgical patients，es－pecially coagulase-negative staphylococci，however，G- pathogens has increased recently，especially acinetobacter baumannii，therefore nosocomial meningitis pathogens should be considered in the choice of empirical antibiotic treatments.

Abstract:Objective：To evaluate the risk factors for post traumatic cerebral infarction（PTCI） in patients with moderate or severe traumatic brain injury（TBI）. Methods：A retrospective review of 327 moderate or severe TBI patients admitted to our department between January 2010 and December 2011 was performed. Patients who had a history of cerebral infarction，severe heart disease，poorly controlled hypertension，hyperlipidemia and diabetes，and Glasgow coma scores（GCS） of more than 12 were excluded to avoid interfering factors. Patients who died during the first 3 d after the admission were also excluded. All cases were analyzed in aspects of gender，age，GCS，basal skull base fracture，traumatic subarachnoid hemorrhage（tSAH），cerebral hernia，low blood pressure or shock，decompression craniotomy. Statistical analysis was performed using chi-square and student’s t test. Multivariate analysis was per－formed using logistic regression to identify independent risk factors for the development of PTCI. Results：There were 37 cases（11.31%） suffering from PTCI. Attack rate of PTCI for patients with GCS 3-8 was 16.11%（34/211） while for patients with GCS 9-12 was 2.59%（3/116）. High attack rate of PTCI after moderate or severe TBI was also related with basal skull base fracture（19.74%，15/76），tSAH（14.89%，32/215），cerebral hernia（17.95%，14/78），low blood pressure or shock（20.00%，15/75） and decompression craniotomy（14.61%，26/178），with significant correlations（P=0.001，0.031，0.029，0.039，0.001，0.043）.While the patients’ age and gender were not associated with the development of PTCI（P=0.353，0.882）. Conclusions：Low GCS，basal skull base fracture，tSAH，cerebral hernia，low blood pressure or shock，and decompression craniotomy are risk factors for PTCI in patients with moderate or severe TBI. No association is observed between PTCI and gender or increasing age.

Abstract:Objective：To evaluate the diagnostic value of four neuropsychological scales in early diagnosis of Alzheimer’s disease（AD）. Methods：Totally 109 subjects were divided into three groups：24 in AD group，40 in mild cognitive impairment（MCI） group，45 in normal cognition（NC） group. Four scales were adopted including memory and executive screening（MES），auditory verbal learning test-Huashan version（AVLT-H），mini-mental state examination（MMSE）and Montreal cognitive assessment scale（MoCA）. Scores of the four scales were compared and analyzed. Results：For each scale，there were significant differences among the scores of three groups（P＜0.01）. The scores of short-term and of AVLT-H were not correlated with education level（P ＞0.05）. The sensitivity and specificity of MES were 95.6% and 75.0% respectively using a cut-off score of 80. The sensitivity and specificity reached the highest （100.00% and 99.84%） when taking a cut-off score of 4 in long-term delay recall of AVLT-H. Conclusions：Both MES and AVLT-H are of great value in diagnosing AD at early stage and are not influenced by the education level.

Abstract:Objective：To explore the neuromuscular activation（NEURAC） technology in the treatment of children with cerebral palsy and to provide reasonable，standard and effective rehabilitation treatment method for cerebral palsy. Methods：3-D gait exanimation and gross motor function measure（GMFM） evaluation were performed for cerebral palsy children with abnormal gait，poor ability but certain executive capability before the treatment to determine the cause of abnormal gait muscle. Training program was designed and assessment was conducted after training for 1-2 months. Results：（1）Step length was increased（t=-2.352，P=0.033） and step width was decreased（t=2.726，P=0.001） compared with that before the treatment，with statistically significant differences. （2）Total supporting time was （55.15±19.30） ms before the treatment and （60.19±7.79） ms after the treatment（t=-1.031，P=0.319）. Double supporting time was （20.42±17.12）% after the treatment，significantly increased before the treatment of （14.55±7.28）% （t=-1.496，P=0.155）. （3）There was no significant change in swing time and single supporting time.（4）GMFM score after the treatment was obviously improved than before treatment（t=4.99，P=0.001）. Conclusion：Nerve muscle activation technology can improve the walking ability of cerebral palsy children in a comparatively short time.

Abstract:Objective：To study the correlation between serum vitamin A（VA） and blood lipid triglyceride（TG），total cholesterol（TC），low density lipoprotein（LDL），high density lipoprotein（HDL） by researching the levels of VA and lipid among people aged 50 years old or more in gerocomium of Chongqing and to explore the possibly mechanism between VA and lipid. Methods：A total of 480 peo－ple meeting the inclusion criteria were randomly recruited from 12 gerocomiums of Chongqing. Serum VA and blood lipid were mea－sured by high performance liquid chromatography and Siemens 2400 biochemical detector，respectively and their correlation was ana－lyzed. Results：Serum VA was negatively correlated with LDL significantly（β=-0.068，P=0.017） and was not correlated with TC and HDL. There was significant correlation between serum VA and TG（β=-0.338，P＝0.000）. The most significant correlation between serum VA and LDL was seen in 70-80 years old（β=-0.136，P=0.005），and VA was a protective factor for the prevalence of high LDL（OR=0.286，95%CI＝0.110－0.747）. Conclusions：VA is significantly correlated with LDL but not with HDL and TC in gerocomium of Chongqing. The most significant correlation between serum VA and LDL is seen in 70-80 years old group. There is significant corre－lation between VA and TG，the mechanism of which should be studied further.

Abstract:Objective：To study the knowledge，attitude and behavior of Chongqing rural residents and to provide advices for launching control and prevention for chronic disease. Methods：Totally 2 406 residents aged 18 years old and above in rural areas of Chongqing were randomly selected out using multi-stage stratified random sampling method. Results：The awareness rates of hypertension and di－abetes were 53.6% and 23.1%. There were significant differences in scores of attitude towards prevention and control measures between patients and non patients（t=-2.21，-2.26，-2.45，-2.13，-2.12，all P<0.05）. Smoking and drinking rates were 24.8% and 8.4%，repst－cively. 36.0% and 23.0% residents investigated tested blood pressure and blood glucose in recent 6 months while 8.8% and 27.4% had the awareness of weight control or actively consulted knowledge of prevention and control. Conclusions：Low knowledge level，weak consciousness and low rate of health behavior formation in rural residents suggest that health education should be taken according to different people and intensity of chronic diseases prevention and control should be increased.

Abstract:Objective：To prepare SPIO-PLL-pshRNA molecular probes and to preliminarily evaluate the feasibility of transfecting SKOV3 cells and MR imaging in vitro. Methods：Superparamagnetic iron oxide modified with Poly-L-lysine（SPIO-PLL） was bound with plasmid pGenesil1-shRNA through electrostatic absorption. Concentration of plasmid DNA in the supernatant was detected by micro-ultraviolet spectrophotometer after the centrifugation of the complexes. By this way，the ability of plasmid DNA binding to SPIO-PLL could be evaluated. Conjunction between SPIO-PLL and plasmid DNA was confirmed via gel retardation experiments and zeta potential methods. Cell transfection was evaluated with Prussia blue staining for iron assessment and fluorescence microscope for green fluorescent protein（GFP） expression detection. Changes of cells in MR signal intensities were observed after the transfection. Results：DNA binding tests，gel retardation experiments and zeta potential methods demonstrated that SPIO-PLL could effectively bind plasmid DNA when the w/w ratio of SPIO-PLL and plasmid DNA was more than 6∶1. zeta electric potentials of SPIO，SPIO-PLL and SPIO-PLL-pshRNA were （-14.8±0.35），（15.2±0.55） mV and （3.6±0.35） mV，with statistical differences among groups（F=6323.782，P=0.000；LSD：all P=0.000）. After transfection with the SPIO-PLL-pshRNA complexes，iron particles could be detected in cells with Prussia blue staining；green fluorescence could be observed by fluorescent microscope；MR imaging showed T2*WI signal intensities reduction（558±19，427±16，283±19 and 191±8 in non transfection group，10，20 μg/ml and 40 μg/ml groups respectively；600±36 and 670±16 in agarose group and distilled water group）. The signal intensity was decreased with the increase of complex concentration and statistical differences were observed among groups（F=279.667，P=0.000；SNK-q：all P<0.05）. Conclusions：SPIO-PLL nanoparticles can effectively bind with plasmid pGenesil1-shRNA as SPIO-PLL-pshRNA molecular probe. This probe can successfully transfect SKOV3 cells and T2*WI signal intensity reduction can be observed by MR scanning.

Abstract:For the first 30 years since its discovery，reversible protein acetylation has been studied and understood almost exclusively in the context of chromatin remodeling and gene transcription accompanying the research of histone acetyltransferase（HAT），histone deacetylase（HDAC） and HDAC inhibitors（HDACI）. With the discovery of non-histone acetylated proteins and acetylation-modifying enzymes in cellular compartments outside the nucleus，the regulatory potential in diverse cell progressing of reversible acetylation has slowly been recognized in the last decade. However，the scope of protein acetylation involvement in complex biological processes re－mains uncertain. The recent development of new technology has enabled，for the first time，the identification and quantification of the acetylome，acetylation events at the whole-proteome level. These efforts have uncovered a stunning complexity of the acetylome that potentially rivals that of the phosphoproteome. The remarkably ubiquitous and conserved nature of protein acetylation revealed by these new studies suggests the regulatory power of this dynamic modification. The establishment of comprehensive acetylomes will change the landscape of protein acetylation，where an exciting research frontier awaits.

Abstract:Objective：To produce polymethylmethacrylate（PMMA） electrospun nanofibers with different topographic features and to test their potential as scaffold after neural injury. Methods：Electrospining was used to produce nanofibers with random and aligned topographic features，PC12 cells was cultured on nanofibers and PMMA film was served as control to reveal the respective neurite growth manner on different substrate. Results：Both the random and aligned PMMA electrospun nanofibers had fine surface character and there was no difference in fiber diameter detected（t=1.079，P=0.474）. PC12 cell could successfully adhere，grow and differentiate on each kind of substrate，and on aligned nanofibers the neurite which were also aligned as the orientation of the fibers were significantly longer than those cells cultured on random fibers and film（q=35.36，P=0.006；q=29.23，P=0.009）. Conclusion：Aligned PMMA electrospun nanofibers has the potentiality as scaffold after neural injury.

Abstract:Objective：To construct the eukaryotic expression vector containing exogenous miR-134 precursor，to study its expression in the human oligodendroglia（OL） cells and to provide the technical platform for further studying the function of miR-134 and its target gene. Methods：miR-134 precursor sequence was amplified from the human genomic DNA and cloned into pEGFP-N1. After being confirmed by PCR and sequencing，recombinant plasmid was transfected into human OL cells and transcription level of miR-134 was detected by real-time PCR. In addition，transcription efficiency was monitored by the inverted fluorescence microscopy. Results：The target bands were consistent with the expected results by enzyme digestion and PCR identification，and the inserted sequence was correct in gene sequencing detection. After the recombinant plasmid being transfected into OL cells，fluorescence quantitative PCR detection showed that there was no difference between pEGFP-N1 empty plasmid group and control group in the expression level of miR-134（P=0.882）. Moreover，the miR-134 expression level of pEGFP-miR-134 plasmid transfected group was 52 times over that of pEGFP-N1 empty plasmid group，with significant statistical differences（P=0.023）. Conclusions：The miR-134 expression vector is successfully constructed，which paves the way for the future experiment.

Abstract:Objective：To construct recombinant adenovirus vector，which takes thyroid-stumulating hormone receptor（TSHR） A sub－unit（TSHR289） as immunogen，to construct Grave’s animal model and to explore traditional Chinese medicine prescription for the prevention and treatment of Grave’s disease. Methods：TSHR289 gene was amplified from the plasmid of pSV2neoECE-TSHR289-6H-dhfr to attB1-Kozak-TSHR289/6xHis/IRES/eGFP-attB2 vector with overlapping PCR. PCR products purified by agarose gel electrophoresis were collected and adenovirus vector pAV.EX1d-TSHR289/6xHis/IRES/eGFP（pAd-TSHR289/6xHis） was recombined with GatewayR technology. Then recombined adenovirus vector was translated into Escherichia coli Stbl3；positive clones were screened out by PCR before sequencing. pAd-TSHR289/6xHis was transferred into HEK293A cells to produce recombinant adenovirus；aden－ovirus was purified by CsCl gradient centrifugation and titer of the virus was measured by TCID50. Then mRNA expression of of TSHR289 in HEK293 was measured by RT-PCR. Grave’s model was created by immunizing BALB/c mice with Ad-TSHR289/6xHis. Results：Construction was proved correct by screening positive bacterial. Enhanced green fluorescence protein and cytopathic effect of HEK293A cells was observed by microscope after infection. RT-PCR demonstrated that mRNA expression of TSHR289 in recombinant adenovirus transferred cell were 285 times higher compared with those of untransfected cell. The titer of the virus was 3.16×1010 PFU/ml and the positive rate of T4 and TRAb was 7/10 on the 10th week after BALB/c mice being immunized. Conclusions：Recom－binant adenovirus vector of overexpressed human TSHR289 can be successfully constructed. The animal’s experiments prove that Ad-TSHR289/6xHis can induce autoimmunity and produce TRAb，which provide the prerequisite for the experiment in vivo of treating Grave’s disease.

Abstract:Objective：To determine the three dimensional structure and to analyze the molecular mechanism of staphylococcus aureus（S.aureus） infection for developing a new antibacterial drugs by cloning，expressing，purifying and crystallizing the surface protein SdrE from S.aureus. Methods：SdrE gene was amplified and cloned into the vector. Recombinant plasmid，pW28-SdrE，was trans－formed into E.coli B834 to express SdrE protein. SdrE protein was purified by Ni2+-NTA affinity chromatography column and DEAE anion-exchange chromatography column. State of aggregation in solution was analyzed by Hiload Superdex 200 gel flittration column. SdrE crystal was screened with Hampton kit and crystal was optimized with chessboard method. Results：（1）Recombinant plasmid pW28-SdrE was constructed and expressed in soluble form in E.coli B834 strain. （2）Puried SdrE exhibited as monomer in solution. （3）Single crystals of SdrE with good shape and diffraction capability were obtained. Conclusions：SdrE crystals can be grown by classic protein purification and crystal cultivation methods，which provides an important base for structural and functional study of SdrE in future and is necessary for developing new antibacterial drugs.

Abstract:Objective：To develop an analytical method based on high performance liquid chromatography（HPLC） for the determination of azodicarbonamide（ADC） in flour. Methods：Samples were extracted with acetone，degreased with n-hexane，then separated with a agilent Eclipse XDB-CN column（250 mm× 4.6 mm，5 μm） using acetonitrile-ammonium acetate（20 mmol/L） as mobile phase with a gradient elution program and were detected with diode array detector with the wavelength of 245 nm. Results：ADC had a good linear relationship within the range of 1.0-90.0 mg/kg；the correlation coefficient was higher than 0.99 and the limit of quantification（S/N≥10） was 5.0 mg/kg. Average recoveries of ADC at spiking levels of 5.0，10.0，45.0 mg/kg were ranged from 68.6% to 116.9% and relative standard deviations were less than 7.0%. Conclusions：The method developed is simple，rapid，accurate and reliable；it can be used for the determination of ADC in flour.

Abstract:Objective：To investigate the clinical significance of plasma free metanephrines（MNs） in the diagnosis and prognosis of pheochromocytoma by high performance liquid chromatography-electrochemical detection（HPLC-ED）. Methods：HPLC-ED was carried out to detect plasma free MNs，including 21 patients with pheochromocytoma，26 healthy volunteers，23 patients with primary hypertension and 24 patients with adrenal mass but without pheochromocytoma. Results：Plasma concentrations of free MNs in patients with pheochromocytoma were significantly increased compared with those of other groups（P<0.05）. Taking normetanephrine（NMN） 150 ng/L and（or） metanephrine（MN） 120 ng/L as cut-off value，the sensitivity and specificity for diagnosis of pheochromocytoma were 100% and 89% respectively. Plasma concentrations of free MNs showed significant positive relationships with tumors size in patients with pheochromocytomas（NMN：r=0.952，P=0.000；MN：r=0.510，P=0.018）. Plasma concentrations of free MNs in patients with pheochromocytoma dropped rapidly after the surgery. Conclusion：Plasma free MNs are good markers for the diagnosis and progno－sis of pheochromocytoma.

Abstract:Objective：To establish an ultra performance liquid chromatography（UPLC） method for detecting content of uridine，guanosine and adenosine in Kangbingdu oral liquid. Methods：ACQUITY UPLC BEH C１8（1.7 μm，2.1 mm×50 mm） was used and the mobile phase was methanol and water by gradient elution mode；column temperature was 30 ℃，at a flow rate of 0.5 ml/min and detection wavelength of 254 nm. Results：The linear range of uridine was 2.033-20.33 μg/ml and the regression equation was as follows：Y=80 664X-1 046（r=0.999 9）. The linear range of guanosine was 1.248-12.48 μg/ml and the regression equation was as follows：Y=55 261X-742（r=0.999 9）. The linear range of adenosine was 1.254-12.54 μg/ml and the regression equation was as follows：Y=52 155X -480（r=0.999 9）. The average recovery rates were 98.56％（RSD=1.21％），98.77％（RSD=1.09％） and 98.88％（RSD=1.22％） respectively. Conclusions：The method is accurate，rapid and reliable and can be used to determine the nucleoside compounds in Kangbingdu oral liquid.

Abstract:Objective：To investigate the effect of different down-regulation protocols on the outcome of in vitro fertilization-embry－otransfer in patients with endometriosis. Methods：Clinical data of 108 patients with endometriosis who underwent in vitro fertilization-embryo transfer in the reproductive center of the Third Affiliated Hospital of Zhengzhou University from January 2010 to December 2012 were analyzed． According to the different down-regulation protocols，patients were divided into two groups：prolonged therapy group（n=33） and super prolonged therapy group（n=75）. Outcome of assisted reproduction between the two groups was analyzed. Results：There was no obvious difference in patients’ age，infertility duration，basic of follicle-stimulation hornone，luteinizing hormone and estradion levels（P >0.05）. Total duration and dosage of gonadotropin were obvious higher in super prolonged therapy than in pro－longed therapy group，but the number of oocytes and cleavage，number of transferable embryo and number of high quality embryo were fewer in super prolonged therapy than in prolonged therapy group（P<0.05）. Clinical pregnancy rate and embryo implantantion rate were higher in super prolonged therapy than in prolonged therapy group（P<0.05）. Conclusions：Use of super prolonged protocol regulation can improve embryo implantantion rate and pregnancy rate for patients with endometriosis at Ⅲ-Ⅳ stage.

Abstract:Objective：To evaluate the efficacy of fludarabine combined with cyclophosphamide（FC） regimen for chronic lymphocytic leukemia（CLL）. Methods：Clinical data of 41 patients with CLL treated by FC regimen were retrospectively analyzed. Results：The rates of complete response（CR），partial response（PR） and overall response（OR） were 41.5%，36.6% and 78.1% respectively. There was no difference between initial and secondary treatment groups in CR，PR and OR rates（58.8%，23.5%，82.4% vs. 29.1%，45.8%，75%，P ＞0.05）. The main side effects were myelosuppression and immunologic function suppression and there was no difference in hematotoxicity between two groups（P ＞0.05）. The overall sur－vival（OS） and disease-free survival（DFS） rates of 32 patients who had finished 4 cycles of chemotherapy，were 59.1% and 35% respectively.The DFS rate of initial treatment group was better than that of secondary treatment group（53.6% vs. 20.8%，P ＜0.05），but there was no difference in OS rate between two groups（77.1% vs. 47.3%，P ＞0.05）. Conclusions：FC combined chemotherapty regimen can improve the prognosis and disease-free survival of patients with CLL. It is the currently preferred scheme for patients with CLL due to its good efficacy and tolerance，but more attention should be paid to the long-term effect.

Abstract:Objective：To investigate the effect of continuous femoral nerve block on the deep vein thrombosis（DVT） in patients under－going total knee arthroplasty（TKA）. Methods：During February 2011 to March 2013，115 patients undergoing TKA were recruited in this study. These patients were assigned randomly into 2 groups：study group（n=59） and control group（n=56）. All these patients received unilateral TKA under intravenous plus inhalation anesthesia. After the surgery，patients in study group were treated with continuous femoral nerve block for the postoperative pain，meanwhile，patients in control group were treated with patients controlled intra－venous analgesia（PICA）. The level of D-dimer in plasma was detected and the ultrasonic detector was applied to estimate the DVT. Visual analogue scores（VAS） at 2，6，12，24，48，72 h after the surgery were recorded. Functional exercise was assessed with the distance of patients’ walk within 3 d after TKA. Results：After the TKA，the plasma level of D-dimer was significantly lower in study group than in control group（P=0.000 8，P=0.000，P=0.000）. The outcomes of the ultrasonic detection showed that 6 out of 56 patients in control group developed DVT after TKA，whereas 2 out of 59 patients in study group developed DVT after TKA（?字2=4.296，P=0.039）. The VAS at each time point was lower in study group than in control group. The distance was longer in study group than in control group（P=0.000，P=0.000，P=0.000）. Conclusions：Continuous femoral nerve block has better effect on DVT after TKA than PICA，which may relate with its better analgesic effect and its promotion of postoperative functional exercise.

Abstract:Objective：To present the results of a prospective study using a combined 585 nm pulsed dye laser（PDL）/1 064 nm neodymi－um：yttrium-aluminum-garnet（Nd：YAG） laser to treat steatocystoma. Methods：One hundred and two patients were enrolled in this study with 45 non-inflammatory lesions and 57 inflammatory lesions. The parameters were set as follows：energy densities of 5.0-8.0 J/cm2 with PDL and 10-30 J/cm2 with 1 064 nm Nd：YAG laser；pulse duration of 0.5 ms with 585 nm PDL and 15 ms with 1 064 nm Nd：YAG laser. Delaying time between 2 beams was medium and spot size was 7 mm. Results：The curative rates for inflammatory steatocystoma were 89.5%，96.5% and 98.2% in 2 weeks，4 weeks and 24 weeks respectively while those for non-inflammatory steato－cystoma were 88.9%，95.6% and 95.6% in 2 weeks，4 weeks and 24 weeks respectively，without differences between the two（P=1.000） Two non-inflammatory lesions relapsed with a recurrence rate of 1.96% during 6-18 months’ telephone follow-up. Transient purpura was present in all patients and was faded away within 1-3 weeks. The incrustation was observed in 43.1% patients. The severity of pain was assessed by visual analogue score（VAS） and the average score was 2.41±0.97. However，other adverse effects were limited. Conclusions：Combined 585 nm PDL/1064 nm Nd：YAG laser is a valuable treatment option for steatocystoma. It can eliminate both inflammatory and non-inflammatory steatocystoma with good healing and minimal adverse effects.

Abstract:Objective：To analyze the three dimensional characteristics of condylar position between centric relation（CR） and maximum intercuspation（MI） in patients with mandibular deviation and to discuss its influence on orthodontic treatments of dental maxillofa－cial deformities. Methods：Thirty-four patients with mandibular deviation were taken as experimental group and fifty patients with non-deviation as control group. Experimental group was then divided into two groups according to the existence of temporomandibular joint disorder（TMD）. CR/MI occlusions and occlusal interferences of CR position were recorded respectively. Condylar positions of CR and MI were measured respectively by measures condyle displacement（MCD） and the proportion of occlusal interferences and CR-MI discrepancy，as well as the offset in condylar position between CR and MI in three dimensional direction were compared. Results：（1）Percentages of occlusion interferences in experimental and control group were 94.12% and 72% respectively with statistical differences（χ2=6.421，P=0.010） and the occlusion interferences of both groups were located at the posterior teeth，accounting for 91.18% and 68% respectively；（2）Percentages of CR-MI discrepancy in experimental and control group were 79.41% and 12.00% respectively（ χ2=38.59，P=0.000）；（3）In the experimental group，there was no significant difference in the percentage of CR-MI discrepancy（ χ2=0.167，P=0.687） and the amount of displacement of CR-MI in vertical（t=0.283，P=0.779） or sagittal（t=-0.743，P=0.463） plane between deviated and non-deviated side（P >0.05）. There was no significant difference in the amount of displacement of mandible and ＣＲ-ＭＩ discrepancy in three dimensional direction between TMD group and non-TMD group（P >0.05）. Conclusions：Patients with mandibular deviation show a higher tendency to suffer occlusal interferences and CR-MI discrepancy，so it is necessary to analyze the consistency of CR-MI in patients with mandibular deviation and to make a treatment plan in the ideal position of jaw.

Abstract:Objective：To study and to summarize the clinical feature，imaging feature，key points for diagnosis and differential diagnosis of bronchioloalveolar carcinoma（BAC） so as to improve the clinicians’ understanding，diagnosis and treatment of BAC. Methods：Clinical data of 32 patients with BAC in the department of respiratory and department of thoracic surgery in our hospital from May 2010 to September 2013 were studied retrospectively，including clinical manifestation，thoracic image，laboratory test and pathological examination，etc. Results：Clinical features of BAC were cough，expectoration，blood in sputum，progressive dyspnea；a lot of white frothy sputum was the remarkable characteristic of BAC，but not the common performance. The imaging of BAC was of 3 types：solitary nodular type，pneumonia type and diffusing type，each type had its typical imaging findings. Thirty-two patients were diagnosed as BAC by pathological or cytological examination. Conclusions：Mastering the clinical feature，imaging feature，key points for diagnosis and differential diagnosis of BAC and performing early lung biopsy are contribute to diagnosing BAC at early stage and reducing the rate of misdiagnosis and mistreatment significantly.

Abstract:Objective：To investigate the effects of the gentle liver manipulation，including toching，retracting and moving liver lobes gently，on hepatic ischemia-reperfusion injury during hepatic ischemia and its mechanism. Methods：Thirty SD rats were divided into three groups randomly：sham-operated group（sham group），ischemia reperfusion group（IR group：produced by total inflow occlusion for 15 min） and manipulation group（MN group：induced with 13 min gentle manipulation including touching，retracting and moving lobes during 15 min ischemia）. Serum aminotransferases alanine transaminase and aspartate aminotransferase（ALT and AST），tumor necrosis factor-α（TNF-α） and Pselection in the liver were measured after reperfusion. Results：In MN and IR groups，serum ALT and AST（P=0.000） as well as pathological injury scores（P=0.000） were increased；TNF-α and P-selection expressions were higher（P=0.000） compared with those in sham group. Those variables mentioned above were significantly increased in MN group compared with those in IR group（P=0.000）. Conclusions：Gentle in vivo liver manipulation increases the reperfusion injury during hepatic is－chemia，which may be caused by the decrease of pro-inflammatory and activity of neutophils.