Objective To investigate the status of vitamin(Vit)B12 deficiency in elderly inpatients in department of neurology.Methods 827 patients admitted to the department of neurology of Shanghai Punan hospital from March 2007 to July 2008,aged 60 and older,with no severe hepatosis and renal disfunction,and without using VitB12 preparation for 3 months,were included in the study.The levels of serum VitB12,folate,homocysteine(Hcy) were tested to screened patients with VitB12 deficiency.The results of symptoms,positive signs of neurological system and the laboratory examination were analysed and compared between patients with or without VitB12 deficiency.Results There were 827 patients,with the mean age of(77.1?7.5)years old,fulfilled the enrolling criteria.VitB12 deficiency was found in 163 patients(19.7%).The prevalence was higher in female(21.9%)than male(17.4%),which had a tendency of increased with age.In the patients with VitB12 deficiency,the rate of complicating gastrointestinal diseases was higher.Compared with patients with normal serum VitB12 level,there were more cases suffered from unstable gait(27.0%),hypopallesthesia(34.4%),and more cases combined with some chronic diseases such as cerebral infarction(62.6%),hypertension(68.7%),coronary heart disease(39.9%).77% cases had neuroelectricphysiological abnormalities,but only 9.82% cases had megaloblastic anemia.Conclusions VitB12 deficiency is common in elderly inpatients in neurology department,whose clinical manifestations are various and atypical,and the neurological symptoms are more common than megaloblastic anemia.

Aim To investigate the neuroprotective effects of osthole(Ost)on the primary cultured cortical neurons transfected with APP595/596 gene and its underlying mechanism.Methods Neonatal mouse cortical neurons were transfected with APP595/596 gene to establish AD cell models for the further study.Then,the cell viability was detected by CCK-8 assay,and the leakage of lactate dehydrogenase(LDH)was assayed by LDH kit to evaluate the injury degree.Transferase-mediated nick end labeling(TUNEL)was used to evaluate the cell apoptosis.The expression of β-amyloid peptide(Aβ)and β-site APP cleaving enzyme 1(BACE1)was measured by immunofluorescence,while the miRNA-107 was measured by RT-PCR.Results Compared to model group,Ost could significantly improve the neurons viability,decrease the LDH release and prevent the apoptosis.Ost also inhibited the expression of Aβ and BACE1 at protein level,while enhanced the expression of miRNA-107 at gene level.Conclusion Ost plays a neuroprotective role in neurons transfected with APP595/596 gene in part through up-regulating miRNA-107.

Objective To establish an experimental animal model of hypertriglyceridemic(HTG)panereatitis by using special lipoprotein lipase(LPL)deficient HTG heterozygous mice.Methods LPL deficient HTG heterozygous mice and wild type mice were divided into experiment group and control group,then each group was subdivided into 12 h,24 h subgroup.Acute pancreatitis(AP)was induced by caerulein intra-abdominal injection for 7 times(50μg/kg)at the interval of 1 h;the mice in the control group received same amount of normal ssline.The serum level of TG,amylase was determined,and morphologic changes were scored.Results The serum level of TG in LPL deficient HTG heterozygous mice was(3.55±0.27)mmol/L,which was significantly higher than that of wild type mice[(0.94±0.18)mmol/L,P＜0.05].The serum level of TG and amylase in heterozygous mice at 12 h was(3.55±0.27)mmol/L and(3685±484)U/L,which was significantly higher than those in wild type mice[(0.92±0.11)mmol/L and(2501±410)U/L,P＜0.05].The pancreatic tissue edema,necrosis,bleeding and inflammatory cell infiltration score was 3.94 ±0.21,3.94±0.21,1.84±0.25 and 1.84±0.25 in heterozygous mice,which was significantly higher than that of wild type mice(3.06±0.01,2.52±0.51,0.46±0.22 and 0.58±0.38,P＜0.05).Conclusions The serum level of TG was moderately and stably elevated in heterozygous mice.The mice developed more severe panereatitis after caemlein induction.which was an ideal experimental model for study of mechanism of HTG pancreatitis.

Objective To assess the reliability and validity of measurement instrument of nurses' humanistic practice ability (MIHPA).Methods By means of literature analysis,interviewing with experts and two rounds of Delphi with 17 nursing management and nursing education experts,we got preliminary evaluation questionnaire.680 nurses from three level of first-class hospital were selected by convenient sampling,who took part in the survey on nurses' humanistic practice ability.After two weeks,we re-measured 50 nurses.Such methods as exploratory factor analysis,confirmatory factor analysis and reliability analysis were used to screen and assess the questionnaire entries.Results The exploratory factor analysis indicated that the measurement instrument of nurses' humanistic practice ability was composed of five principal components,the five principal components could explain 65.137％ of the total variance.Confirmatory factor analysis showed x2/df=2.158,CFI=0.952,NFI=0.915,RFI=0.901,IFI=0.953,TLI=0.944.Cronbach's α of the total scale was 0.933 and retest reliability was 0.822.Reliability of each subscales was 0.782～0.867 and retest reliability was 0.782～0.848.Conclusions The measurement instrument of nurses' humanistic practice ability,which contained 5 dimensions and 23 items,has good reliability and validity.

Objective: To investigate the prevalence of hyponatremia and the relationship between hyponatremia and prognosis of dilated cardiomyopathy (DCM) for in-hospital patients. Methods: A total of 515 consecutive in-hospital DCM patients treated in HF center of Fu Wai Hospifal from 2008-10 to 2013-10 were retrospectively studied. Hyponatremia was deifned as the serum level of sodium < 135 mmol/L at ifrst admission. The prevalence of hyponatremia and the relationship between hyponatremia and DCM prognosis were studied including the risk of in-hospital time and mortality, the rates of all cause death and HF worsening death after discharge. Surviving patients were followed-up by clinical or telephone visit until 2014-11 or until the death. Results: There were 134/515 (26.0%) patients suffered from hyponatremia at admission, the serum level of sodium was related to HF symptom duration, NYHA functional classiifcation, systolic blood pressure (SBP), left atrial diameter and total bilirubin level, allP<0.01. Compared with non-hyponatremia, the patients with hyponatremia presented longer in-hospital time(14.8±11.1) days vs (11.2±5.8) days and higher in-hospital mortality (18.7% vs 1.8%), bothP< 0.01. There were 483 survivors discharged and were followed-up for (30.7 ± 19.5) months, during that period, the rates of all cause death and HF worsening death were 26.5% and 21.9% respectively. The patients with hyponatremia had the higher rates of all cause death (47.7% vs 20.3%) and HF worsening death (44.0% vs 15.5%), bothP<0.01. Multiple Cox regression analysis showed that with adjusted HF history (> 6 months vs≤ months ), NYHA functional classiifcation (Ⅱ-Ⅳ), SBP (per 10 mmHg elevation), total bilirubin level (per 1 mg/dl change) and LVEDD (per 5 mm change), the hyponatremia at admission is still one of the important independent predictors for all cause death (HR=1.836, 95% CI (1.248-2.702),P<0.01 and HF worsening death HR=2.139, 95% CI (1.406-3.253),P<0.01 in DCM patients after discharge. Conclusion: Hyponatremia is a common electrolyte disorder for in-hospital DCM patients, it is related to longer in-hospital time and higher mortality; higher rates of all cause death and HF worsening death after discharge in DCM patients.

AIM: To explore the role and possible mechanism of polyamine in L - arginine inhibiting cardiac hypertrophy induced by isoproterenol (ISO). METHODS: Hypertrophic model of rats was established using ISO. Pretrea-ted with L - arginine, hypertrophy status of rats was determined by hypertrophy coefficient, collagen content and the expression of ANP mRNA. High performance liquid chromatography ( HPLC) was used to measure the concentrations of polyamines. Western blotting was performed to detect the expressions of ornithine decarboxylase ( ODC) and spermidine/ spermine Nl - acetyltransferase (SSAT). The activity and levels of NOS and NO in serum were also observed. RESULTS : Hypertrophy coefficient and expression of ANP mRNA increased significantly after injection of ISO for 7 d. Moreover, cardiac muscle fibres became thick and disorganized. Pretreated with L - arginine, the above index decreased. Meanwhile , the concentration of polyamine was decreased and plasma NO content and NOS activity were increased, the expression of ODC was downregulated and the expression of SSAT was upregulated. CONCLUSION: Exogenous L - arginine inhibits cardiac hypertrophy through downregulating L - arginine/polyamine pathway and upregulating L - arginine/NO pathway.

Aim To investigate the effects of neurotro-phin-3 ( NT-3 ) gene overexpression on the differentia-tion into cholinergic neuron of neural stem cells ( NSCs) in vitro and its underlying mechanism. Meth-ods Brain-derived NSCs from newborn mice were iso-lated and cultured in vitro and determined by immuno-fluorescence. The NSCs were divided into three groups: NSCs, GFP-NSCs and NT-3-NSCs groups. The expression of NT-3 was detected by immunofluo-rescence and ELISA. Then, the ability of NSCs on dif-ferentiation into cholinergic neuron was detected by im-munofluorescence and RT-PCR, and the Acetylcholine Assay Kit was used for acetylcholine ( ACh) , and the expression of Hes1 , Mash1 and Ngn1 mRNA was de-termined by RT-PCR. Results The neurosphere dis-played Nestin and Sox 2-postive by immunofluores-cence, suggesting that the cultured cells were NSCs. The proportion of ChAT immunopositive cells was sig-nificantly higher in the NT-3-NSCs group than that in the other two groups ( P <0. 01 ) . Ach secretion in NT-3-NSCs was significantly elevated compared with the other two groups ( P <0. 01 ) . NSCs transfected with NT-3 increased the levels of Mash1 and Ngn1 mR-NA, and decreased the level of Hes1 mRNA ( P <0. 05 ) . Conclusion NT-3 can significantly promote the in vitro differentiation of NSCs into cholinergic neu-rons via probablly inhibiting Notch signaling pathway.

Aim To investigate the effects of osthol on cell apoptosis and inflammatory cell infiltration after brain stab wound injury in mice. Methods The mice underwent the stab wound injury by a needle, then were randomly divided into sham operation group, model group, osthol 10, 20, 30 mg · kg-1 treatment group. The main examinations included mice brain wa-ter content; the apoptotic cytokines Bax, Bcl-2, Caspase-3 mRNA expression were assessed by PT-PCR; immunohistochemistry staining was used to de-tect neutrophils (MPO) and microglia (Iba-1) infiltra-tion and Caspase-3 positive cell expression around in-jured lesions. Results Treatment with osthole 20, 30 mg·kg-1 group significantly reduced the water content in injured brain, improved the ratio of Bax/Bcl-2, and reduced the expression of apoptosis cytokine Caspase-3 mRNA. Osthole 30 mg·kg-1 treatment group obvious-ly reduced the infiltration of neutrophils and microglial cells and significantly reduced the number of apoptotic cells around the injured cerebral cortex. Conclusion Osthole has therapeutic effect on stab wound injury in mice, and the possible mechanism may be by reducing the infiltration of inflammatory cells and reducing apop-totic cells.

Aim To investigate the effect of osthole on neuron synapses infected APP gene and its underlying mechanism. Methods The neurons were divided into three groups:GFP, APP, APP+Ost groups. The neu-rons were infected APP gene with containing mutational site in vitro for mimicking the characterstics of Alzhei-mer’ s disease ( AD) . The cell viability was assessed by CCK-8 , the expression of synapsin-1 was deter-mined by immunofluorescence, and the concentration of PSD-95 and SYP were detected by ELISA. The ex-pressions of Aβ1-42 , CAMKK2 , phoshorylated AMPKα1 , AMPKα1 protein were determined by West-ern blot. Results Strong APP staining was visible in neurons infected with APP and abundant expression of Aβ1-42 , a neurotoxic oligomer. Compared with APP group, APP+Ost group significantly increased cell vi-ability, promoted the expression of synapsin-1, up-reg-ulated the concentration of PSD-95 and SYP, and de-creased the expressions of CAMKK2 and p-AMPKα1 . Conclusions Ost can protect the neuron synapses a-gainst infected with APP gene. Its neuroprotective effect may be related to inhibiting the CAMKK2/AMPK signal pathway.

Objective To investigate the effect of intramuscular and oral mecobalamin on hematologic markers and neurologic signs in elderly patients with vitamin B12 deficiency.Methods One hundred and twenty-six patients with vitamin B12 deficiency who fulfilled the inclusion criteria were randomly divided into 3 groups,42 cases in each,including the intramuscular group (to receive mecobalamin therapy intramuscularly),the oral group ( to receive mecobalamin therapy orally) and the control group ( without mecobalamin therapy).The changes of hematologic markers including hemoglobin (Hb,g/L),mean corpuscular volume (MCV,fl),serum levels of vitamin B12 (ng/L),folate and total homocysteine (Hcy,μmol/L),and neurological signs before and after treatment were compared among these groups.Results Baseline characteristics among the three groups were similar.After a 6-month therapy,there were no differences in any markers in control group patients in comparison to baseline;for patients in the intramuscular group,the blood vitamin B12 levels increased from (139.13 ± 31.57)ng/L to (328.10 ± 42.35 )ng/L (P ＜ 0.001 ).Hcy levels decreased from (36.29 ± 16.23 )μmol/L to ( 18.23 ± 9.85 ) μmol/L ( P ＜ 0.001 ).Hb rose from ( 125.34 ± 16.21 ) g/L to ( 132.79 ± 15.98 )g/L (P =0.037).MCV reduced from (92.98 ±5.35)fl to (87.65 ±5.74)fl (P ＜0.001 ) ;For patients in the oral administration group,the blood vitamin B12 levels increased from ( 138.19 ± 29.95) ng/L to (487.79 ±32.21 ) ng/L ( P ＜ 0.001 ).Hcy levels decreased from ( 33.27 ± 11.51 ) μmol/L to ( 17.49 ± 10.13 ) μmol/L( P ＜ 0.001 ).Hb rose from ( 125.89 ± 17.65 ) g/L to ( 133.46 ± 16.26) g/L ( P =0.041 ).MCV reduced from (93.08 ± 5.10 ) fl to ( 89.29 ± 5.37 ) fl ( P =0.001 ).After the 6-month therapy,there were somewhat improvement in MMSE scores of the intramuscular ( 28.24 ± 3.89 vs.27.85 ± 3.56,P =0.633 ) and the oral groups (27.97 ± 3.77 vs.27.34 ± 3.15,P =0.408 ) compared with baseline,but the differences were not significant.The achilles tendon reflex of the intramuscular ( 1.86 ± 0.67 vs.1.56 ± 0.61,P =0.035 ) and the oral groups ( 1.79 ± 0.64 vs.1.43 ± 0.51,P =0.006 ) were enhanced compared with baseline.Foot vibration sensation of the intramuscular ( 1.35 ± 0.37 vs.1.06 ± 0.41,P =0.001 ) and the oral groups ( 1.24 ± 0.52vs.1.01 ± 0.43,P =0.03) were improved compared with baseline.After treatment,the serum VitB12 concentration in the oral group were higher than that of the intramuscular group ( P ＜ 0.001 ).There were no significant differences in other indexes between the two groups.Conclusion Mecobalamin,administered either intramuscularly or orally,may improve the hematologic markers and neurologic signs in elderly patients with vitamin B12 deficiency.