Objective To investigate the changes in cortical excitability and inhibitory circuits of patients with Alzheimer's disease (AD) or behavioral variant frontotemporal dementia (bvFTD) using transcranial magnetic stimulation (TMS).Methods Forty-four patients with AD,30 patients with bvFTD and 44 healthy controls were enrolled in the study.The epidemiological data of all subjects were collected.Each participant received a neurological evaluation and neuropsychologic tests which included Mini-Mental State Examination (MMSE),Activities of Daily Living (ADL) and Hamilton Rating Scale for Depression (HAMD).Neurophysiological evaluations including resting motor threshold (rMT),active motor threshold (aMT) and cortical silent period (CSP) were conducted by means of TMS.Neurophysiological parameters were compared among groups.Results There were significant differences in MMSE,ADL and HAMD assessments among 3 groups,but no significant differences between AD and bvFTD groups.There were significant changes in left rMT(46.52％ ± 8.77％,52.00％ ± 7.30％ and 52.14％ ± 8.27％,F =6.295,P=0.003),left aMT(29.68％ ±7.01％,35.13％ ±8.46％ and 35.39％ ±7.24％,F=7.735,P=0.001) and right rMT(47.82％ ±7.94％,52.07％ ±8.77％ and 53.12％ ±8.61％,F=4.772,P=0.010) among 3 groups.AD patients showed significantly reduced left rMT and aMT comparing to bvFTD patients (P =0.017 and 0.008 respectively) and controls (P =0.005 and 0.002 respectively).In addition,AD patients had a significant decrease in right rMT comparing to controls (P =0.011).There were no statistically significant differences in TMS parameters between bvFTD patients and controls.Conclusions AD is associated with hyperexcitability of the motor cortex,whereas the lack of changes in motor cortical excitability is found in bvFTD.TMS technique may be helpful in differential diagnosis between AD and bvFTD.

Objective To investigate the clinical characteristics of myeloperoxidase (MPO)-antineutrophil cytoplasmic antibody (ANCA) associated vasculitis (MPO-AAV) with pulmonary injury and the relationship between pulmonary injury and ANCA against light chain of MPO (LCMPO-ANCA).Methods A total of 195 patients with newly diagnosed primary active MPO-AAV were recruited in this prospective study.Indirect immunofiuorescence assay was used to detect peri-nuclear ANCA (p-ANCA).Immunoblotting and ELISA were used to detect myeloperoxidase-antineutrophil cytoplasmic antibody (MPO-ANCA).Clinical features of patients with both positive p-ANCA and MPO-ANCA were collected.Disease activity was evaluated by Birmingham Vasculitis Activity Score-version 3 (BVAS-V3) Recombinant light chain of MPO was used to coat substrate of LCMPO-ANCA by ELISA.The clinical characteristics of pulmonary injury and its correlation with serum levels of p-ANCA,MPO-ANCA and LCMPO-ANCA were explored.Results All 195 patients (64 male and 131 female),consisted of 191 patients (98.0％) with microscopic polyangiitis,3 patients (1.5％) with granulomatosis with polyangiitis,and 1 (0.5％) with eosinophilic granulomatosis with polyangiitis including 64 men and 131 women.Their mean age was (63.2 ±13.5) years old.The level of MPO-ANCA had a positive correlation with general BVAS-V3 (r =0.193,P =0.007) in all patients,and the level of LCMPO-ANCA was positively related with the pulmonary BVAS-V3 (r =0.228,P =0.001).As for multiple systemic damages,the incidence of lung involvement was 60.51％(118/195),which ranked second to renal involvement (71.80％,140/195).The most common pulmonary injuries represented as pulmonary infiltration of 80.51％ (95/118),pleural effusion / pleurisy of 41.53％(49/118),pulmonary nodule or cavity of 22.03％ (26/118).Compared with those without lung involvement,the patients with pulmonary injuries were older [(66.39 ± 10.70) years old vs (58.30 ±15.72) years old;t =4.277,P =0.001],had a shorter course of disease [2.00(1.00,10.50) months vs 3.00(1.00,3.50) months;t =-2.283,P=0.024],and higher scores of general BVAS-V3 (18.21 ±6.08 vs 15.18 ± 5.64;t =3.501,P =0.001).Also,in the patients with pulmonary lesions,the positive rate of LCMPO-ANCA was significantly higher (35.59％ vs 6.49％;x2 =21.569,P ＜ 0.001),and the level of LCMPO-ANCA was significantly higher (0.377 ±0.229 vs 0.285 ±0.079;t =3.399,P =0.001)than those without lung involvement.The pulmonary BVAS-V3 in the patients with LCMPO-ANCA was significantly higher than that in the patients without LCMPO-ANCA (4.34 ± 2.10 vs 2.59 ± 2.52;t =4.301,P ＜ 0.001),whereas the pulmonary BVAS-V3 was not correlated with LCMPO-ANCA (r =0.035,P =0.708) in patients with lung injuries.Conclusion Pulmonary injury was relatively common and insidious in patients with MPO-AAV.To monitor ANCA level is necessary in patients with pulmonary injury.LCMPO-ANCA might play an important role in the pathogenesis of pulmonary lesions in AAV.

Objcctive To detect the telomerase activity in rat hepatic oval cells, and to explore the relationship between telomerase expression and the proliferation and differentiation of oval cells. Methods The 2-acetamidofluorene/partial bepatectomy (2-AAF/PH) rat model was used to induce the proliferation of oval cells. Oval cells were isolated by modified collagenase perfusion and gradient centrifugation. Electron microscope exami-nation and immunofluorescence were adopted to identify oval cells. Immunohistochemistry, RT-PCR and fluores-cence quantitative PCR were used to detect the expression of telomerase in oval cells. All the data collected were analyzed by t test. Results The proliferatiun of oval cells was successfully induced by 2-AAF/PH rat model. Freshly isolated oval cells showed a large and ovoid nuclei, a small proportion of cytomplasm and a cobblestone appearance. When viewed by electron microscopy, there were few and immature organelles, and the nucleus/ cytoplasm ratio was high. Immunofluorescence staining showed that oval cells expressed OV-6, alpha-fetoprotein, cytokeratin-19, albumin and c-kit. Telomerase reverse transcriptase (TERT) was located in the nuclei of oval cells which were around the portal areas. As oval cells differentiated into small hepatocytes, the number of TERT-positive cells decreased significantly. The expression level of TERT mRNA in normal rat liver tissue was 2.27 times higher than that in LE-6 oval cells; the expression level of TERT mRNA in the isolated oval cells was 1.26 times higher than that in LE-6 oval cells. The telomerase activity decreased gradually (from △A=1.05, 1.15 to △A=0.25, 0.45) as the increase of oval cells passage (from passage 24 to passage 40) (t=17.74, 12.38, P<0.05). Conclusions Oval cells have telomerase activity. Telomerase may be indispensable for maintaining the proliferative and multi-directional differentiation abilities of oval cells.

Objective Investigate the prognosis of patients undergoing liver transplantation (LT) for end-stage autoirnmune liver disease (ALD).Method The clinical data of 45 patients with endstage ALD undergoing LT from April 2001 to March 2015 in the first affiliated hospital of Zhejiang University were analyzed retrospectively.The postoperative cumulative survival rate of the recipients was calculated,and the causes of death were analyzed.The postoperative rejections,new onset viral hepatitis and ALD recurrence were also analyzed.Result In 45 ALD recipients,33 cases survived and the postoperative 5-year cumulative survival rate was 78.8％.Causes for 12 dead cases were mnultiple organ failure,liver graft failure,respiratory complications,hemorrhage and hepatic artery embolization.In 45 ALD recipients,6 cases suffered rejection after operation with the incidence bing 13.3％.One case suffered new onset hepatitis B infection 8 years after opcration.One recipient suffered primary disease (primary biliary cirrhosis) recurrence 2 years after operation,and 1 recipient with primary disease (primary sclerosing cholangitis) developed into overlap syndrome.They all survived for a long term after active treatments.Conclusion Most LT recipients with endstage ALD can obtain a long term survival.Attentions should be paid to the immunosuppressive regimens in early period after LT,prevention of infection,rejection and postoperative new onset viral hepatitis,and timely diagnosis of primary disease recurrence.

Objective To investigate the expression of CD55 and myeloperoxidase (MPO) on neutrophils in patients with MPO-specific anti-neutrophil cytoplasmic antibody associated vasculitis(MPO-AAV), and analyze the relationship between the expression and clinical manifestation.Methods Forty untreated patients with active MPO-AAV (patient group) and 30 healthy volunteers (control group) were enrolled in this study.The CD55 on neutrophils and both membrane and cytoplasmic MPO were detected by flow cytometry.Serum fragment-from the activated complement factor B(Ba) and MPO were measured by ELISA.The clinical activity of vasculitis was valued by Birmingham vasculitis activity score-version 3(BVAS-V3).The significance of laboratory data was evaluated by Spearman correlation test and multivariate linear regression analysis.Results (1)The mean fluorescence intensity(MFI) of CD55 expressed on neutrophils was significantly higher than that in control group[4 068.6±2 306.0 vs 2 999.5±1 504.9,P=0.033].Similar results of serum MPO and Ba in patient group were found compared to controls [500.0(381.0, 612.7) IU/L vs 286.9(225.5, 329.1) IU/L,P<0.001;35.2(25.2, 79.5) ng/L vs 18.0(15.0, 28.0) ng/L,P<0.001], respectively.However, MIF of cytoplasmic MPO in patients was significantly lower than that of control group(1 577.1±1 175.9 vs 3 105.3±2 323.0,P=0.003).(2) In patient group, cytoplasmic intensity of MPO was negatively associated with the serum levels of MPO(r=-0.710,P<0.001) and Ba (r=-0.589,P=0.001).Moreover, serum MPO was positively associated with serum Ba(r=0.691,P<0.001).Membrane intensity of CD55 on neutrophils was positively correlated with patient age (r=0.514, P=0.001), C reactive protein (r=0.376, P=0.018), peripheral neutrophils count (r=0.485, P=0.001) and BVAS-V3 (r=0.484, P=0.002), whereas negative correlation between membrane CD55 and disease duration was seen (r=-0.403,P=0.01).(3) The result of multiple linear regression analysis showed there was statistically significant positive correlation between MFI of CD55 expressed on neutrophils and BVAS-V3 (β=0.001,P=0.027).Conclusions In MPO-AAV,CD55 expression on neutrophils is markedly enhanced, which is one of the independent risk factors related to disease activity.It might protect neutrophils from attacking AAV, CD55 expression on neutrophils is markedly enhanced, which is one of the independent risk factors related to disease activity.It might protect neutrophils from attacking by complement alternative pathway.Activated neutrophils release more MPO and lysosome to intensify the inflammation reaction and aggravate the disease.Thus CD55 might become a new potential target for the treatment of this disease in the future.

As a member of G protein coupled-receptors superfamily, free fatty acid receptor 1 (FFAR1), is also known as GPR40, has been shown to regulate numerous pathophysiological processes in a variety of tissues and organs. The activated FFAR1 has a variety of biological functions. For instance, it can not only regulate metabolism of fatty acids and glucose, but also play an important role in immune inflammatory response, it may be a potential drug target for the treatment of various chronic inflammatory diseases. In this review, we focus on the recent researches of FFAR1's action in the regulation of pathophysiological processes, its molecular mechanism and new agonists development. At the same time, this review will take the discovery of series FFAR1 agonists as examples, and display the applied prospects of FFAR1.

Objective To construct expression vectors of calmodulin(CaM)mutants N2 and C2,and to express,purify,and identify the mutant proteins,in order to study the interactions between CaM and calcium channels. Methods The cDNA of N?lobe and C?lobe of CaM were used to prepare the cDNA of N2 and C2. Next,the recombinant cDNAs were cloned into a pGEX?6p?3 plasmid,and the recombinant plasmids were trans?ferred into E.coli BL21 cells. The transfected BL21 cells were stimulated with IPTG. The fusion proteins were extracted by ultrasonication and puri?fied by using GS?4B beads. Finally,protein activity was identified by the pull?down assay. Results Both the restriction digestion map and the DNA sequence identification results confirmed that the recombinant plasmids were successfully constructed. SDS?PAGE results showed high purity and concentration of N2 and C2 proteins. Their activities and binding abilities with the calcium channel fragment were confirmed by the pull?down assay.Conclusion In this study,expression vectors of N2 and C2 are successfully constructed,and physiologically active N2 and C2 CaM mutant proteins are obtained.

BackgroundWith the increasing prevalence of dry eye and the continuous improvement of living standards,the problem of dry eye more and more get the attention of people.At present,China still lacks the large population-based epidemiological data of dry eye. Objective To investigate the prevalence and possible risk factors of dry eye in a community of Huidong County of population aged 14 and over.Methods From September 2010 to January 2011,using questionnaires and examination of dry eye related,2800 people were selected randomly for cross-sectional survey.Those suspected as dry eye were examed by the SchirmerⅠtest ( S Ⅱ T),tear-film breakup time(BUT),corneal fluorescein staining(F1).Results In the 2475 questionnaire effectively,154 persons were diagnosed as dry eye,and the prevalence rate of dry eye was 6.22％,8.06％in females,4.14％in males.The prevalence rate increases with age.The S Ⅰ T and BUT decreased with increasing age.S Ⅰ T and BUT in females are less than males.Foreign body sensation is the primary complaints of patients.Logistic analysis showed that the most common risk factors in dry eye are age and gender.The system disease and eye diseases,eye fatigue and long exposure to dust are also main determinants.ConclusionsThe population prevalence rate of dry eye increased with age,the prevalence rate of dry eye in females is higher than that in males.The key factors associated with dry eye are age,gender,systemic disease and eye diseases,occupation,working environment.

Objective To investigate the effects of siRNA-mediated knockdown of S100A4 expression on the inva?sion and migration of SNB19 glioma cells. Methods The S100A4 expression was knockdowned using S100A4 siRNA in SNB19 glioma cells. Glioma cells were assigned into control group,siRNA-negative control treated group (siRNA-NC) and siRNA-S100A4 group. RT-PCR and western blot were used to detect the mRNA and protein expression of S100A4, respectively. The wound-healing assay and transwell invasion assay were used to determine the ability of migration and invasion of SNB19 glioma cells, respectively. The expression of matrix metalloproteinase 9 (MMP-9), matrix metallopro?teinase 2 (MMP-2) and E-cadherin proteins were evaluated by using western blot. Moreover, the morphology of lamellipo?dia of glioma cells were examined by using inverted phase-contrast microscopy. Results The mRNA and protein expres?sion levels of S100A4 was obviously down-regulated after transfection of S100A4 siRNA. Compared with control group, the mRNA expression levels of S100A4 in siRNA-NC group and siRNA-S100A4 group were 0.97±0.07 and 0.21±0.04,respectively(P<0.01). The protein expression levels of S100A4 in control, siRNA-NC and siRNA-S100A4 groups were 78.12%±2.63%, 77.16%±3.00%and 37.95%±2.71%, respectively(P<0.01). The migration and invasiveness capability were decreased up to 46% and 55% in the siRNA-S100A4 group compared with the control group(P<0.01). The pro?tein expression levels of MMP-9 and MMP-2 were inhibited up to 62% and 68%(P<0.01)whereas the expression of E-cadherin was increased up to 154%(P<0.01)in the siRNA-S100A4 group. The lamellipodia became smaller or unex?tended in siRNA-S100A4-treated SNB19 glioma cells. Conclusion S100A4 plays an important role in the invasion and migration of glioma cells, suggesting that S100A4 might be a potential candidate for anti-glioma strategy to prevent the invasion and migration of glioma cells.

Objective To summarize surgical treatment of Takayasu arteritis,and analysis the drug treatment effect during the perioperative period.Methods Retrospective analysis 46 patients with Takayasu's arteritis disease and received cardiovascular surgery between January 2010 to December 2015,in Anzhen Hospital.By collecting their clinical characteristics,preoperative drug therapy,surgical treatment,pathological examination results to analyze operation conditions,effect of drugs and preoperative conditions.Results The perioperative mortality rate was 2.2％ and the complication rate was 23.9％ in 46 patients.There were 34 patients with symptomatic relief in the perioperative period,11 patients didn't take hormone drugs before operation.There were 11 cases of complications during the perioperative period,of which 7 patients were in active stage and 10 patients had not been used before operation.Conclusion The surgical treatment of patients with Takayasu's arteritis disease can effectively improve symptoms.The patients in Takayasu's arteritis active stage will affect the outcome of the surgery.Rational use of hormone drugs before surgery,can effectively control the patient's condition,improve the rate of remission of symptoms,and effectively reduce the incidence of perioperative complications.