Objective To find a sensitive index and to investigate the working memory impairment of patients with amnestic mild cognitive impairment.Methods Fifteen patients with amnestic mild cognitive impairment,15 healthy matching aging controls were performed a matching-to-sample task while event-related potential(ERP),reaction time and correct rate were recorded.Subjects were required to press a button in the match condition and another button in the conflict condition.Results In the matching condition,there was no difference between the two groups in distribution(parietal lobe),peak latency and amplitude of P300(F_ 1,28 =1.0324,P=0.3183;F_ 2,42 =0.543,P=0.585).In the conflict condition,the distribution of N270 was fore head,and its latency of patient group were more delayed than the aging group(F_ 1,28 =25.3264,P=0.000),but its amplitude showed no significant changes(F_ 1,28 =0.507,P=0.482).The result of brain mapping showed same change.Conclusion The N270 component is more sensitive than P300 to reflect the central executive function impairment of working memory in patients with amnestic mild cognitive impairment.

Objective To study the protective effect and molecular mechanism of trypsin inhibitor on reperfusion injury after cerebral ischemia. Methods The model of ischemia for 1h and repufusion for 24h of rat cerebrum was set by ligating MCAO described by zea-longa. 24 male rats were divided randomly into the sham operation group, the control group and trypsin inhibitor group. The presence of neurological function deficit was measured by Zea-Longa method, and the immunohistochemical staining and TUNEL were used to detect p53 protein expression and cell apoptosis in the brain tissues respectively. Results The score of neurological function deficit was zero in the sham operation group, 2 8?1 0 in the control group and 1 3?0 7 in trypsin inhibitor group. There were 12 3?2 5 p53 immunostaining positive cells in the control group and 5 5?1 3 in trypsin inhibitor group. The number of apoptotic cells was zero in the sham operation group, 7 6?1 0 in the control group, and 3 5?0 9 in trypsin inhibitor group. There was a significant difference in all above observing indices between the control group and trypsin inhibitor group(P

Objective To establish the murine congenital infection model by MCMV and observe the pathological changes and infection status of brain tissue.Methods After anesthesia,mice who were pregnant 11-13.5 days (E11-13.5 d) were intra-amniotic injected one uterus by one with virus (MCMV K181 suspension,1 μl,1×103 PFU).The control group of the same period was intra-anmiotic injected with culture medium DMEM (1 μl).Carefully reset the uteruses and close the abdomen.After 5 days of separated feeding,kill the pregnant mice,take the fetus out of the uterus,anesthetize and kill them.Make frozen sections of these fetal brains.Some sections were stained using conventional HE method,to observe the pathological changes under the light microscope.Detect MCMV early antigen in the brain tissue by immunohistochemistry staining and immunofluorescence assay.Results The survival rates of the infected group were 71.9％.Compared with the control group,intra-amniotic inoculation of MCMV does not affect the rate of fetal survival,fetal absorption,fetal death and the average weight of the heads,but decrease their average weight of the bodies.The pathological changes are found in the brain tissue of the mouse in the infection group.Through enzyme immunohistochemistry assay,there are many MCMV infected cells in brain-ventricular zone,brain subependymal zone,cerebral cortex and hippocampus area in the infection group.Similar findings were observed by immunofluorescence method.Conclusion By intra-amniotic injection of MCMV suspension,murine model of MCMV congenital infection can be successfully established.This model could be used to study the mechanisms of encephalodysplasia caused by congenital CMV infection in vivo.

Objective To investigate the values and characteristics of 75 g oral glucose tolerance test (OGTT) in pregnant women.Methods A total of 6 103 singleton pregnant women aged (30.4±3.8) years (18-49 years) who delivered in Peking University First Hospital between May 1,2011 and December 31,2012 underwent the 75 g OGTT at gestational age of 24-28 weeks.They were divided into five groups based on maternal age:＜25 years (n=222,3.6％),25-years (n=2 485,40.7％),30-years (n=2 573,42.2％),35-years (n=683,11.2％),and ≥ 40 years (n=140,2.3％).The normal values of the fasting,1 h and 2 h blood glucose were lower than 5.1,10.0 and 8.5 mmol/L.Gestational diabetes mellitus (GDM) was diagnosed when blood glucose of any point was higher than or equal to normal value.Comparison between groups was tested by analysis of variance and LSD test.Logistic regression was used to calculate the risk for GDM in different age groups.Results (1) The fasting,1 h and 2 h blood glucose levels were in Gaussian distribution.The （-x）+2s were 5.51,11.12 and 9.49 mmol/L.The 97.5 percentile were 5.63,11.32 and 9.95 mmol/L.Fasting plasma glucose of ＜ 25,25-,30-,35-,and ≥ 40 years were (4.53±0.40),(4.60±0.40),(4.67±0.43),(4.74±0.46) and (4.82±0.49) mmol/L.The 1 h blood glucose were (6.98± 1.70),(7.55± 1.60),(7.92± 1.63),(8.30± 1.71) and (8.76± 1.86) mmol/L.The 2 h blood glucose were (6.11±1.33),(6.53±1.27),(6.89±1.33),(7.23±1.50) and (7.57±1.60) mmol/L.Therewas statistical difference in the blood glucose levels at a same time-point test among different age groups (F=29.61,60.17 and 72.29,all P＜0.01).(3) The total prevalence rate of GDM was 21.1％ (1 290/6 103) ; and the prevalence rates were 9.9％ (22/222),16.7％ (414/2 485),22.7％ (583/2 573),32.1％ (219/683) and 37.1％ (52/140) among the five age groups,respectively,with significant differences (x2=120.68,P=0.00).Compared with the group aged ＜25 years,the OR (95％CI) of the prevalence among 25-,30-,35-,and ≥40 years group were 1.82 (1.16-2.86),2.66 (1.70-4.18),4.29 (2.69-6.86) and 5.37 (3.08-9.39),respectively.Conclusions Advanced age is a risk factor for GDM.The risk of GDM increases significantly after 35 years old and pregnancy in women aged ＜ 35 years can reduce the risk of GDM.

Objective:Positive effects of bioactive glass (BG) on proliferation,mineralization,and differentiation of human dental pulp cells (hDPCs) was already verified in various former studies.The Arg-Gly-Asp-Ser sequence (RGDS) was confirmed of affecting cell adhesion.Before further investigation,the objective of this study is to investigate whether RGDS can affect the effects of BG on the adhesion,proliferation and mineralization of hDPCs.Methods: hDPCs were harvested from third molars of 18-25-year-old individuals after informed consent.Enzyme digestion technique was used.The 4th to 6th ge-neration of hDPCs were used for all experiments.The cells of the experimental groups were cultured in Dulbecco minimum essential medium (DMEM) containing ionic dissolution products of BG and RGDS of seve-ral concentrations (12.5 mg/L,25.0 mg/L,50.0 mg/L,100.0 mg/L,200.0 mg/L).DMEM containing ionic dissolution products of BG without RGDS was used for cell culture as control group.Cell adhesion was tested 4 h after cell seeding by MTT assay.Cell proliferation was examined at 1,3,5,7,and 9 d after cell seeding by MTT assay.Cell mineralization was investigated on days 14 and 28 by alizarin red staining.After being stained and dried,mineralized nodules were dissolved by cetylpyridinium chloride (CPC) for semi-quantitative test.Results were statistically analyzed by one way ANOVA,SPSS (version 19.0) and P<0.05 was considered to be significant.Results: Cell adhesion in BG group showed no difference from that in DMEM group.Compared with BG group,hDPCs in BG+RGDS groups suggested weaker cell adhesion.When the concentration of RGDS increased,the adhered cell number decreased.hDPCs cultured with BG and RGDS showed lower proliferation activity in the early stage,while no significant difference was observed after 3 d.BG group promoted the mineralization of hDPCs compared with positive control group,negative control group and RGDS group.No significant difference was observed between BG+RGDS group and BG group or between RGDS group and positive control group.Conclusion: BG promotes proliferation and mineralization without affecting cell adhesion of hDPCs.Unbounded RGDS inhibits cell adhesion,but has no influence on the positive effects of BG on the proliferation and mineralization of hDPCs.

Objective To compare the advantages and disadvantages of three methods of yeast-like fungi identification of vulvovaginal candidiasis (VVC).Methods The pathogenic candida strains were identified by CHROM agar chromogeinc culture medium,the chromogenic medium of Autobio and VITEK 2 system,and the result of VITEK 2 system was the gold standard.Results All candida strains were appraised.By VITEK2 system,217 candida isolates were identified as candida albicans,and 26 were detected as non-candida albicans.214 isolates were identified as candida albicans and 13 isolates were identified as non-candida albicans by Autobio.214 isolates were appraised as candida albicans,16 isolates were identified as non-candida albicans.The coincidence rate of identification of CHROMagar and the chromogenic medium of Autobio were 94.7％ and 95.06％.Compared with VITEK 2 system,there was no significant difference between these two chromogeinc culture medium identifying candida albicans and non-candida albicans.Conclusion The chromogenic medium of Autobio has higher cost-effectiveness,higher color discrimination and simpler technical operation,and is worthy of promotion in identifying candida species in clinic.

Objective To explore the effects of Buyang Huanwu decoction(BYHWD)on expressions of nuclear factor-κB p65(NF-κBp65)and its inhibitor( I-κB)in signal transduction of NF-κB in brain tissue of rats with focal cerebral ischemia injury. Methods 180 Sprague-Dawley(SD)rats were randomly divided into normal group,sham-operated group,model group,pynolidine dithiocarbamate(PDTC)group,minocycline(MC)group and BYHWD treatment group,each group 30 rats. The rats of PDTC group were given PDTC 100 mg?kg-1?d-1 by intra-peritoneal injection. In MC group,MC was given by filling the stomach,the dose was 2.35 g?kg-1?d-1,the drug solution was prepared by adding the distilled water,and the total volume of drug solution to fill the stomach was kept at the same volume in various groups,thus the concentration of the drug was different. In BYHWD group,BYHWD was given,the dose was reduced to 5 g?kg-1?d-1 according to the body surface area dose conversion formula about people and animals. In sham-operated group and model group,the distilled water was given in the same volume as other drug solution. The protein expression levels of NF-κBp65 and I-κB in ischemic tissues were examined by using immunohistochemical method on the time points 7,14 and 21 days after treatment in each group. Results Compared with model group, the cell numbers with expression of NF-κBp65 in PDTC group,MC group and BYHWD group were significantly decreased along with the prolongation of therapy time,the decrease in number was more and more,until 21 days,it reached the valley level(cell/400 times HP:44.00±6.91,45.33±6.55,18.67±2.14 vs. 126.00±5.78,all P<0.05);the number of cells with expression of I-κB was obviously increased,the differences being statistically significant(all P<0.05),but the differences in expression of NF-κBp65 among the treatment groups at the different time points were not statistically significant(all P>0.05). After treatment for 7 days,the number of cells with positive expression of I-κB protein in BYHWD group was less than that in MC group(cell/400 times HP:55.00±3.40 vs. 72.50±4.29,P<0.05);after treatment for 14 days,the number in BYHWD group was approximately the same as that in the MC group, the difference being not statistically significant(93.50±6.15 vs. 93.00±6.20,P>0.05),and after treatment for 21 days,the number in BYHWD group was significantly higher than that in MC group(88.83±4.95 vs. 71.17±7.16, P<0.05). Conclusion BYHWD can regulate the expressions of inflammatory cytokine I-κB and NF-κB in signal transduction of NF-κB in ischemic brain tissue to inhibit the inflammatory reaction,thus it has the protective effect on cerebral ischemia.

Objective To investigate the nature of Th17 cells in murine cytomegalovirus(MCMV)infection during the acute stage,we characterized the frequency of IL-17A-producing CD4 T cells and the level of Th17 cytokines,IL-17A,in MCMV-infected mice.Methods BALB/c mice were randomly divided into two groups.One was infected with MCMV Smith for establishing disseminative infection; the other was sham-inoculated control.On day 3,7,14 and 28 of the experiment,three mice of each group were randomly chosen to be killed separately.Real-time PCR was used to detect MCMV loads in organs of MCMV-infected mice,the pathology of spleen was observed by HE staining.The frequency of CD4+IL-17A+ T cells in total splenocytes of mice was detected by flow cytometry.The level of IL-17A in culture supernatants of splenocytes was measured by double antibody sandwich ELISA.Results MCMV loads in salivary gland reached the peak on day 14 after MCMV infection,the most severe spleen injury was also shown on day 14,the frequencies of CD4+IL-17A+ T cells in total splenocytes increased significantly( all P＜0.01 ) in MCMV-infected mice than those in controls,and reached the peak on day 14 ( 1.14％ ±0.09％ vs 0.19％ ±0.04％,t =17.551,P=0.000).The levels of MCMV-specific IL-17A in culture supernatants were increased dramatically in MCMV-infected mice than those in controls on day 14 [ (81.98± 12.37) pg/ml vs (44.96±8.44)pg/ml,t=4.281,P=0.006].In MCMV-infected mice,correlation was positive between the levels of MCMV-specific IL-17A in culture supernatants and MCMV loads in salivary gland tissues (r=0.54,P＜0.05 ),the levels of IL-17 A in culture supernatants were higher in more severe spleen injury.Conclusion Thl7 cells and IL-17A were involved in the immunity response during acute MCMV infection.They may correlate with the persistence of MCMV and the pathology of spleen in infected mice.

ObjectiveTo observe the changes of AIM2 ( absent in melanoma 2) inflammasome during early murine cytomegalovirus (MCMV) infection.MethodsBALB/c mice were randomly divided into two groups.One was infected with MCMV Smith for establishing disseminated infection,the other was sham-inoculated control.On days 1,3,5 and 7 of the experiment,three mice of each group were randomly chosen to be killed separately.The expression of AIM2,ASC and caspase-1 in splenic macrophages was detected by Western blot,the levels of IL-1β and IL-18 in sera were measured by double antibody sandwich ELISA,and the viral titers in salivary gland tissues were quantified by a standard plaque assay.Results The MCMV titers in salivary gland tissues were gradually increased in MCMV-infected mice on days 3,5 and 7,while the expressions of AIM2 in macrophages were began to increase on day 1 and significantly increased and reached the highest level on day 3 but gradually decreased afterwards.The relative intensity of AIM2 on day 3 differed significantly between the MCMV-infected mice and the controls (1.121±0.243 vs 0.240±0.046,P＜0.01,t test),as did ASC ( 1.318±0.333 vs 0.248±0.090,P＜0.01 ) and caspase-1 ( 1.085±0.243 vs 0.247±0.064,P＜0.01 ).Meanwhile,the levels of IL-1β and IL-18 in MCMV-infected mice were (112.72±5.20) pg/ml and (42.74±4.23) pg/ml,and the levels were significantly higher (P＜0.01 ) than those in controls [ (47.86±4.35) pg/ml and (22.60±2.82) pg/ml].ConclusionThese results demonstrate that AIM2 inflammasome is activated in macrophages during early MCMV infection and could be as a therapeutic target for CMV-induced diseases.

Objective To explore the correlation between the expression of IL-17A and the degree of spleen damage in acute mouse cytomegalovirus(MCMV) disseminated infection in vivo and to understand the mechanism about how IL-17A involved in the pathological damage of the spleen in MCMV infection.Methods An acute disseminated MCMV infection model was established in mice.BALB/c mice were randomly divided into two groups.Mice in group one were infected with MCMV Smith to establish disseminated infection.Mice in another group were sham-infected control.Three mice from each group were randomly chosen to be sacrificed on days 3,7,14 and 28 after the infection.Viral titers in spleen tissues were determined using a standard plaque assay.The expression of IL-17A mRNA and MCMV mRNA in the splenocytes were measured by RT-PCR.The expression of IL-17A in spleen tissues was observed by immunohistochemical staining.The pathology of the infected mice was assessed by histological examination of H&E stained spleen sections.Results Viral titers and MCMV mRNA in the spleen peaked on day 3,but quickly diminished on day 7.Virus was no longer detectable in the spleen on day 14 after the infection.The expression of IL-17A mRNA was significantly increased during the acute infection and reached the highest level on day 14,then decreased on day 28.It is significantly higher than that of the mock infection group.Immunohistochemistry assay also indicated that the expression of IL-17A in spleen tissue gradually increased to climax on day 14,then decreased on day 28.Accordingly,the pathological damages of spleen tissue in the infected mice deteriorated until day 14,then showed signs of recovery on day 28.The most severe pathological injury of spleen tissue and the highest expression of IL-17A appeared in the same period of time.Conclusion Our results showed a close correlation between IL-17A and the pathological damage in spleen.Thus,IL-17A may contribute to the spleen pathological damage during the acute disseminated MCMV infection.