Aim To explore the effect of N-acetyl-L-cysteine ( NAC ) on β amyloid peptide 25 - 35 ( Aβ25-35 )-induced the increase of calpain activity and its possible mechanism. Methods The activity of cal-pain was induced by 20μmol·L-1 Aβ 25-35 in primary cortical neuron. Neurons were incubated in the absent or present Aβ25-35 , or pre-incubated NAC ( 10 mmol ·L-1 ) , then co-incubated with Aβ25-35 . The meas-urement of calpain activity, H2 O2 level and mitochon-drial membrane potential was performed on a micro-plate fluorometer. The ATP level was detected using a luciferin/luciferase based ATP assay kit. Results In Aβ25-35 treated group, the activity of calpain and H2 O2 was obviously higher than that in control group. How-ever, in neurons pre-incubated in NAC and then co-in-cubated in Aβ25-35 , the calpain activity and H2 O2 level were significantly decreased compared with that in Aβ25-35 group. Upon Aβ25-35 exposure for 12 h, corti-cal neurons showed a significant decrease in mitochon-drial membrane potential and ATP level when com-pared to the control group. Pre-treatment with NAC showed an increase in mitochondrial membrane poten-tial and ATP level as compared to neurons treated with Aβ25-35 alone for 12h. Conclusion This result sug-gests that NAC can attenuate calpain activity induced by Aβ25-35 through protecting mitochondria.

Objective To have a systemic review of the association between relative ratio in proton magnetic resonance spectroscopy (1H-MRS) and Alzheimer's disease (AD). Methods A search in Medline and China National Knowledge Infrastructure (CNKI) was performed,and relevant English and Chinese-language articles about assessing AD with 1 H-MRS were identifed.The data of relative metabolic ratios (NAA/Cr,Cho/Cr,mI/Cr ) from different brain regions (hippocampus,posterior cingulate gyrus,temporal lobe,parietal lobe,frontal lobe,occipital lobe) were extracted from the articles.The quality of the articles was evaluated according to the standard recommended by Newcastle-Ottawa criteria. The Meta-analysis was done with the Review Manager 4.2 to calculate pooled weighted mean difference (WMD) with 95％ confidence interval (95％ CI ),and linear correlation analysis between NAA/Cr ratio and mI/Cr ratio was done by SPSS 17.0.Results Thirty six articles (27 English articles,9 Chinese articles) were included. After heterogeneity test was done,fixed effects model or random effects model was selected. The meta-analysis showed that the NAA/Cr ratio in patients with AD was higher than that in controls ( WMD:-0.14,95％CI:-0.17 to -0.11 ).The mI/Cr ratio in patients with AD was lower than that in controls ( WMD:0.10,95％ CI:0.07 to 0.13 ).There were greatest changes in NAA/Cr ratio and mI/Cr ratio on the hippocampus ( WMD of NAA/Cr: - 0.27,95％ CI:- 0.36 to - 0.19 ; WMD of mI/Cr:0.21,95％ CI:0.10 to 0.33). There were also no differences between patients with AD and controls with respect to the Cho/Cr ratio (WMD:0.01,95％ CI:0.00 to 0.01,P ＞0.05). The NAA/Cr and mI/Cr changes are markedly correlated with each other in different brain regions ( r =0.947,P =0.004). Conclusion The hippocampus region is the first to present neuropathological changes in AD and the changes of NAA/Cr and MI/Cr might reflect the neurodegenerative process of AD.

Objective To evaluate the efficacy and safety of emergency percutaneous coronary intervention(PCI) in patients with non-ST-segment elevation acute coronary syndrome(ACS).Methods A total of 233 patients(emergency group) were treated with emergency PCI within 48 h of heart attack and another 152 patients(delayed group) were treated with PCI after 3-14 days of medical therapy.All culprit lesions were treated.Procedural success rate,the time from admission to angina relief,the length of hospital stay and cardiac events incidence in 30 days were observed.Results The procedural success rates for the emergency group and the delayed group were similar(98.1% vs 95.5%),but cardiac events incidence in 30 days was significantly lower in the emergency group than that of the delayed group(2.9% vs 14.1%,P

Objective To search for the methods of inducing the high stability rat middle cerebral artery occlusion (MCAO) model and to noninvasively assess the method of model effect. Methods Six kinds of filaments with different diameters were used to induce rat MCAO models and their success rate, incidence of subarachnoid hemorrhage, and infarct volume were compared. The model scores were performed to assess the model effect after the operation and at 24 hours after reperfusion. Results Higher model success rate and lower incidence of subarachnoid hemorrhage were achieved when the 0. 28 mm in diameter and 0.32-0.34 mm in tip diameter filaments were used. 1he sensibility and specificity were higher when the model scores were performed at 24 hours after reperfusion. Conclusions Using the filaments of 0.28 mm in diameter and 0. 32-0. 34 mm in tip diameter for intraluminal thread could improved the stability of the models. The model scores at 24 hours after reperfusion could noninvasively assess the model effect.

BACKGROUND: The functional gene fragments integrate into gene vector, which is then transfacted into target cells or joint cavity, through the transgenic target cells continue to secrete a large number of functional gene product, local therapeutic concentrations could be maintained within a long period of time, thus repairing articular cartilage injury. OBJECTIVE: To transfect rabbit articular chondrocytes using recombinant retroviral vector-mediated transforming growth factor-βl (TGFβ_1) in vitro, and to observe its expression and its effect on biological characters of chondrocytes. METHODS: Rabbit chondrocytas were isolated by use of trypsin digestion method. Vector was PLNCX_2 Hind Ⅲ/Not Ⅰ doubly digested and dephosphorylated, connected with some multiple cloning sites and RFP gene following pDsRed_2 double digestion, to build PLNCX_2-RFP. TGFβ_1 gene was amplified from the PGEMT-TGF and connected with PLNCX_2-RFP following double digestion, to build PLNCX_2-TGFβ_1-RFP. Subsequent to packaging retroviral vector, viral supernatant titer was detected. The cultured and transfected chondrocytes in rabbit knee joint were divided into 3 groups: control group (without any transfection), transfected PLNCX_2 group and transfected PLNCX_2-TGFβ_1-RFP group, continued screening 2 weeks to observe the cellular changes. Cell supematant transfected stably were collected for detecting the effect of gene transfection on the chondrocytes with NO detection kit, ELISA assay was applied to determine human TGFβ_1 expression in cell culture supernatant. RESULTS AND CONCLUSION: The recombinant gene PLNCX_2-TGFβ_1-RFP was identified correct sequence by the enzyme digestion sequencing TGFβ_1 and RFP, which showed that the eukaryotic expression vector PLNCX_2-TGFβ_1-RFP had been successfully built as expectation. They were then transfected into packaging calls and cultured, the virus titer was defined as 1×10~6 CFU. Following stable transfection of cartilage cells, red fluorescence can be observed, proving successful transfection. After continuous screening 2 weeks, the scattered adherent calls formed positive clones, and gradually diffusely integrated, cell clusters appeared with common dual cores, the calls proliferated actively. NO concentration in the transfected PLNCX_2-TGFβ_1-RFP group was higher than that of transfected PLNCX_2 group (P < 0.05), no difference was significant between control group and transfected PLNCX_2 group. The control group and the group transfected PLNCX_2 showed no TGFβ_1 expression, while TGFβ_1 concentration was (28.08±3.73) ng/L in the transfected PLNCX_2-TGFβ_1-RFP group. PLNCX_2 ratroviral vector-mediated human TGFβ_1 can be effectively transfected into rabbit knee joint cartilage cells and obtain stable expression, while the transfected cartilage calls proliferate actively.

Objective To investigate the features of the memory function disturbance in patients of generalized seizures and the correlation between the memory function and proton magnetic resonance spectroscopy(1H-MRS).Methods The clinical memory scale of 45 general seizures and 20 normal age and sex-matched controls wss measured and a1 H-MRs was performed in the bilateral hippoeampal.The levels of NAA.Cho.Cr and the ratios of NAA/Cr+Cho were compared between these two groups.Meanwhile,tHe correlation coefficients was calculated and the results of clinical memory scale scoreB and the level of NAA,Cho.Cr and the ratios of NAA/Cr+Cho in groups were compared.Results As compared with the controls.the scores of pointing memory(17.19±5.86),associational learning(20.00±6.77),image free memory(18.44±6.62),association memory of portrait(13.19±6.62)and memory quotient (MQ,79.07±20.20)excepts insignificant figure recognition wcrc significandy decreased in patients of seizures(t=4.794-10.224,all P<0.01);The scores of scale and MQ of both the group of patients who wok antiepileptic drugs(AEDs)and the group of patients whose frequency exceeded 3 per month were significantly lower than the group who didn't take AEDs and the group whose frequency was lower than 3 per month(t=3.267-6.537,all P<0.01).The volumes of hippocampal formations(2.45 ±0.25 on left and 2.56 ±0.31 on right),the level of NAA(12.93 ±1.73 on left and 11.88 ±1.69 on right),Cho(15.02 ±0.86 on left and 14.94 ±0.96 on right),Cr(11.86 ±0.71 on left and 10.71 ±0.42 on right)and the ratios of NAA/Cr+Cho(0.48 ±0.08 on left and 0.39 ±0.07 on riht)in the group of pailents of generalized seizures were significantly difference than those in the controls(t=4.103-5.768,all P<0.01).Patients with seizures showed significant correlation between tlle scores of all items.MQ in clinical memory scale and NAA,NAA/Cr+Cho ratio(r=0.489-0.727,all P<0.01),and inverse correlation between the scores and MQ and Cho of in the bilateral hippocampal(r=-0.604-0.484,all P<0.01).However,there was no correlation between the scores and MQ and Cr of in the bilateral hippocampal.Condusions The patients of the general seizures do have the problem of the disturbance of short-term memory including acoustic memory and some visual memory.Both the high frequency of seizure and AEDs could affect the short-term memory function.The correlation between the scores of all the items and MQ in clinical memory scale proved that lack of nerve cell and hyperplasia of gliocyte was one of the reasons of the short-term memory disorder in epileptic.

Objective To investigate the feasibility and efficacy of vaginal sacrospinous ligament fixation (VSSLF) procedure which reserving uterus in the treatment of uterine prolapse. Methods 8 cases with uterine pro-lapse underwent VSSLF procedures which reserving uterus from July 2006 to December 2008. Those with incontinence d urine simultaneously underwent ventrofixation of the middle piece of the urethra and folding of the ligamentum pos-terius of the uretha and those with anterior or posterior vaginal wall prolapse underwent eolporrhaphia anterior-posteri-or. Results All patients successfully accomplished by VSSLF,without intraoperative or postoperative hemorrhage or organ injury. All patients' prolapse symptoms were relieved efficently according to the pelvic organs prolapse quantifi-cation(POP-Q). Patients were followed-up for 18 months after operation. 8 patients all recovered well without any complication related to the procedure after operation. Conclusion VSSLF is a simple,safe,effective and micro-inva-sire way for patients with uterine prolapse who want to reserve uterus.

AIM:To construct GSK3?-overexpressed SH-SY5Y cells and to observe the effects of GSK3?-overexpression on tau protein phosphorylation and tubulin acetylation in SH-SY5Y engineered cells. METHODS: The cDNA of GSK3? construct was subcloned into mammalian expression vector pBudCE4.1. The integrity of the GSK3? construct was confirmed by sequence analysis. GSK3? was transiently transfected into SH-SY5Y cells using Lipofectamine2000. Western blotting was used to measured protein levels of GSK3? and phosphorylating GSK3?, as well as, the total tau and phosphorylated tau protein and acetylated tubulin. RESULTS: 36 h after transfection, the levels of GSK3? and phosphorylating GSK3? in SH-SY5Y cells were significantly increased compared with non-transfection group and vector group. After 48 h, the levels of phosphorylated tau protein (Ser199/202, Thr231 and Thr205 residues) but not total tau protein were markedly increased in GSK3?-overexpressed SH-SY5Y cells. In addition, the level of acetylated tubulin was lower than that in non-transfection group and vector group. CONCLUSION: The over-expression of GSK3? in SH-SY5Y cells results in robust increases in tau protein phosphorylation at Ser199/202, Thr231 and Thr205 residues, and decreases in tubulin acetylation.

Objective To study the iodine deficiency disorders (IDD) situation in Wenzhou City during 1995-2014.Method According to National IDD Surveillance Project,IDD surveillance had been consecutively carried out during the past 20 years,which consisted of goiter rate in 8-10 years old children,iodized salt and urinary iodine levels.Results The goiter rate of 8-10 years old children was decreased from the highest of 31.09％ (2 190/7 043) in 1995 to the lowest of 2.28％ (77/3 378) in 2014;the highest level of median urinary iodine was 214.78 μg/L,and the lowest level was 74.48 μg/L,and which was increased from 74.48 μg/L in 1995 to 187.00 μg/L in 1996,and then had been maintained at the appropriate level recommended by World Health Organiation (WHO),except that in 1998,2003,2004 and 2006.The qualified rate of iodized salt was increased from 54.95％ (1 471/2 677) in 1996 to 95.52％ (2 548/2 754) in 1999,but decreased to 62.75％ (768/1 224) in 2003,however it was fluctuated from 78.61％ (2 503/3 184) to 92.48％ (2 989/3 232) from 2004 to 2013,and it was 90.43％ in 2014 (2 983/3 300).Conclusions The comprehensive measures for controlling IDD,with universal salt iodization,has been gradually achieved remarkable effect in Wenzhou City,but the non-iodization salt existing in the market is still a problem,and people have misunderstandings about iodization salt.Iodine supplementation had better be conducted according to local conditions and based on scientific policy.

Objective To explore the role of Foxp3+ T cells (Tregs) in liver injury induced by chronic intermittent hypoxia. Methods Thirty-two male Wister rats were divided into four groups:control group (A), high-fat diet group (B), intermittent hypoxia group (C), and high-fat diet and intermittent hypoxia group (D). After 4 weeks, blood samples were collected and livers were surgically removed. Using the standard automatic clinical analyzer to test serum levels of total cholesterol (TC), low density lipoprotein cholesterol (LDL- C), alanina aminotransferase (ALT) and aspartato aminotransferase (AST). The MDA content of liver tissue was measured by colorimetrc method. The levels of TNF-αand IL-1βwere measured by radiommunoassay, and the expression of Foxp3 protein was measured by Western blotting technique. Results Serum levels of TC and LDL-C were significantly higher in B group than those of A, C and D groups, and which were higher in D group than those of A and C groups (P<0.05). There were no significant differences in serum levels of TC and LDL-C between A group and C group. Serum levels of ALT, AST, MDA, TNF-αand IL-1βwere significantly higher in C group than those of A group, and which were significantly higher in D group than those of A, B and C groups (P<0.05). There were no significant differences in these indicators between A group and B group, and between B group and C group. Foxp3 protein expression in liver was significantly lower in D group than that of other groups (P<0.05). Conclusion Foxp3+T regulatory cells involve in the regulation of hepatic injury induced by chronic intermittent hypoxia on the basis of a high-fat diet, and which may play an important role in this process of protective immune response.