In order to set up a base for stem cells to be widely used in clinical medicine, we tried to optimize, in this study, the technique that induces human mesenchymal stem cells (hMSCs) to differentiate into neural stem cells by using cerebrospinal fluid (CSF) from the different groups. After the induction, presence of neural stem cells was confirmed with microscope observation, flow cytometry analysis, immunohistochemistry and fluorescent immunohistochemistry. At the same time, we also compared and analysed the data of the number of stem cells when it totally met the requirements for clinical treatment and the days required. At last, we confirmed that hMSCs could be induced to differentiate into neural stem cells, and that the number of cells totally met the requirements for clinical treatment. But there were some differences both in the number of cells and the days required. Among the groups, the group that marrow mesenchymal stem cells from patients own induced by CSF from healthy volunteers used the shortest time and the quantity of the cells was significantly higher than those of the others.
Cell Differentiation ; Cerebrospinal Fluid ; chemistry ; Culture Media ; chemistry ; Flow Cytometry ; Humans ; Immunohistochemistry ; Mesenchymal Stromal Cells ; cytology ; Neural Stem Cells ; cytology

Objective To evaluate the effect of angiotensin Ⅱ (Ang Ⅱ ) on the change of nephrin phosphorylation both in Ang Ⅱ-infused rat model and cultured podocytes.Methods Thirty Wistar rats were subcutaneously embedded with osmotic minipumps and randomly divided into 3 groups according to receiving either Ang Ⅱ at a dose of 400 ng· kg-1· min-1 or Ang Ⅱ +telmisartan at a dose of 3 mg·kg-1 ·d-1,or normal saline as a control group.Blood pressure and 24-hour urinary albumin were measured at 0 d,7 d,14 d,21 d and 28 d of the experiment.Renal histomorphology was evaluated through electron microscopy.The concentrations of Ang Ⅱ both in blood plasma and kidney were detected by radioimmunoassay.In vitro,cultured murine podocytes were exposed to Ang Ⅱ (10-6 mol/L) pretreated with or without losartan (10-5 mol/L) for different time periods.Nephrin and its phosphorylation expression were analyzed by Western blotting.The distribution of F-actin was presented by FITC-phallodin labeling.The change in phenomenon of F-actin was evaluated by cortical F-actin score index (CFS).Results (1)Ang Ⅱ-infused rats exhibited increased Ang Ⅱ concentration,significant hypertension and marked albuminuria.(2)In Ang Ⅱ-infusion group,nephrin expression was decreased (P＜0.05).Ang Ⅱ-receiving rats displayed diminished phosphorylation of nephrin.(3)In vitro,the phosphorylation of nephrin was significantly reduced after Ang Ⅱ stimulation for 3-6 hours (P＜0.05).(4)Ang Ⅱ stimulatation resulted in irregularly arrangement of F-actin followed by the redistribution of F-actin to podocyte periphery and formation of F-actin ring,in which the CFS obviously increased compared to control (P＜0.05).Conclusions Phosphorylation of nephrin is important for the survival status of podocytes.Ang Ⅱ-induced nephrin dephosphorylation may be an important molecular mechanism for Ang Ⅱ-induced podocyte cytoskeleton rearrangement and foot process effacement.

Objective To investigate the change of serum non-esterified fatty acid (NEFA) level in nondiabetic first-degree relatives of type 2 diabetics, and to explore the related factors in the change.MethodsSerum lipid profile, plasma glucose and insulin levels were measured in 186 type 2 diabetic patients, 565 nondiabetic first-degree relatives of type 2 diabetics and 149 normal controls. Results (1) The fasting NEFA level in first-degree relatives was significantly lower than that of type 2 diabetic patients [(0.53±0.28 vs 0.63±0.31) mmol/L,P＜0.01]and HOMA-IR was significantly higher than that of normal controls (0.98±0.51 vs 0.89±0.47,P＜0.01). (2) The fasting NEFA level in the first-degree relatives with higher body mass index (BMI), plasma glucose or area under curve of glucose concentration (AUCglu) was higher than that in those with lower BMI, plasma glucose , blood pressure or AUCglu (all P＜0.05). (3) NEFA showed significantly positive correlations with BMI, systolic blood pressure, diastolic blood pressure (DBP), AUCglu in the first-degree relatives by correlative analysis (r=0.12, r=0.148, r=0.21 and r=0.281 respectively, all P＜0.05). Stepwise linear regression analysis showed that DBP, AUCglu and age were the independent risk factors of NEFA (all P＜0.01). Conclusion Insulin resistance exists in nondiabetic first-degree relatives of type 2 diabetics, which seems to be related to elevated NEFA levels.

Objective To explore the variation and influential factors of high sensitive C-reactive protein (hs-CRP) level in type 2 diabetic family members. Methods A total of 427 type 2 diabetic patients, 377 non-diabetic first-degree relatives of type 2 diabetics and 135 normal control subjects without diabetic family history were recruited. Serum hs-CRP, clinical and biochemical parameters were measured. The relations among indicators were analyzed. Results Compared with normal control subjects, serum hs-CRP levels in type 2 diabetics and first-degree relatives were significantly increased (both P＜0.05), and the increment was even marked in type 2 diabetics than that in first-degree relatives (P＜0.01). The serum hs-CRP levels in type 2 diabetics and first-degree relatives were positively associated with body mass index, waist-hip ratio, abdominal circumference, postgrandial 2 h plasma glucose, fasting and postgrandial 2 h serum insulin, HOMA-IR, triglyceride, creatinine and negatively correlated with high density lipoprotein-cholesterol. In first-degree relatives, serum hs-CRP level was positively associated with systolic blood pressure and HOMA-β. Conclusion As in type 2 diabetic patients, there exists inflammatory reaction in the non-diabetic first-degree relatives of type 2 diabetics, which may play an important role in the pathogenesis of type 2 diabetes mellitus.

Serum cortisol levels during oral glucose tolerance test (OGTY) were measured in subjects of type 2 diabetic pedigrees. The results showed that cortisol levels during OGTF were higher in type 2 diabetic patients than those in non-diabetic first-degree relatives and normal controls. Fasting cortisol level was positively correlated with fasting plasma glucose level in type 2 diabetic pedigree members. These results suggest that the dysregulation of hypothalamic-pituitary-adrenal axis may coexist in type 2 diabetic patients.

Objective To assess the development,progression and change of nonmotor symptoms in patients with Parkinson' s disease and its impact on patients' quality of life.Methods Eighty-seven consecutive patients with idiopathic Parkinson' s disease were studied.Parkinsonian status was assessed at baseline and 3 years follow-up using Unified Parkinson' s Disease Rating Scale (UPDRS) part Ⅲ & Ⅳ,Nonmotor Symptoms Questionnaire (NMSQuset),Parkinson-related quality of life (PDQ) scales.Paired ttest,Chi-square test,Spearman rank order correlation and hierarchical regression of the major statistical procedures were employed.Results At 3 years follow-up,compared to baseline,the UPDRS Ⅲ score (22.21 ±11.31 vs 30.49± 11.68),UPDRS Ⅳ score(1.00±1.54 vs 2.94±3.12),NMS score (7.98±3.96 vs 12.35 ± 5.12) and PDQ score (28.11 ± 22.88 vs 36.65 ± 26.95) were significantly higher ( t =- 5.54,- 5.75,- 6.46,- 5.29,all P =0.000,respectively).The aggravation of motor and nonmotor symptoms caused the decline of quality of life.The prevalence of constipation,problem of remembenng thing,nocturia ranked tops,and depression,and anxiety were still in the middle,compared with baseline.The prevalence of pains,sweating,dribbling,sense of incomplete emptying etc were significantly increased during the follow-up,△R2 were 21.6％ and 23.4％ respectively,resulting in the deterioration of quality of life.Conclusions PD nonmotor symptoms appear from the early stage.The motor and nonmotor symptoms aggravate over time.

Objective To observe progression of motor symptoms and occurrence of motor complications in parkinsonian patients and investigate the rate of progression of motor symptoms and risk factors of motor complications.Methods One hundred and thirty patients diagnosed with PD in 2007 in Department of Neurology,Xinhua Hospital were followed up for 3 years.The Unified Parkinson' s Disease Rating Scale (UPDRS) and H-Y staging were used to assess and follow up motor symptoms and occurrence of motor complications,and analyze the rate of progression of motor symptoms and risk factors of motor complications with statistics.Results ( 1 ) Mean annual growth in H-Y staging was 2.5％,and UPDRS motor scores was 3.1％ ; the incidence of dysphagia at endpoint in patients was increased by 23.0％ compared with baseline; incidence of falls was increased by 16.7％;(2)Daily levodopa dose at endpoint ( OR =1.004,95％ CI 1.001—1.006,P =0.008 ) was independent risk factors with dyskinesia; While duration ( OR =1.637,95％ CI 1.083—2.473,P =0.019 ),levodopa treatment duration ( OR =0.698,95％ CI 0.494—0.987,P =0.042 ),daily levodopa dose at haseline ( OR =1.005,95％ CI 1.001—1.010,P =0.016) and at endpoint ( OR =1.014,95 ％ CI 1.001 —1.027,P =0.032 ) were risk factors with motor fluctuations.Conclusions As the disease progresses,motor function in parkinsonian patients gradually worsens,the incidence of swallowing difficulty and of falls is increased,and the incidence of motor complications is increased.The total exposure to levodopa in parkinsonian patients is predictor for motor complications.

Objective To derive a questionnaire to assess the risk of developing motor complications through a 3-year prospective investigation on 71 patients of Parkinson' s disease (PD) in the out clinic at our hospital.Methods Three years after the first assessment,71 PD out patients were reassessed using various scales,including Unified Parkinson Disease Rating Scale,Hoehn-Yahr grade,Mini Mental State Exam,Hamilton Depression Scale and Hamilton Anxiety Scale.Results The incidence of motor complications was 43.6％ (31/71).Logistic regression analysis showed that the prognostic factors for motor fluctuation were age of onset ≤ 54 (OR =6.4,95％ CI 1.7-24.5,P =0.006),the occurrence of swallowing difficulty (OR =3.8,95 ％ CI 1.0-14.1,P =0.04) and depression (OR =4.0,95 ％ CI 1.1-13.7,P =0.03),and the prognostic factors for dyskinesia were age of onset ≤54 (OR =48.5,95％ CI 1.9-121.0,P:0.02),the occurrence of falling (OR =64.1,95％ CI 2.9-142.2,P =0.008) and the daily levodopa dosage ＞ 600 mg(OR =17.5,95％ CI 1.1-276.2,P =0.04).Based on the regression model,the assessment questionnaire for motor complicationsincludes the followings:the questionnaire for motor fluctuations:the age of onset ≤54,2 points; the occurrence of swallowing difficulty,1 point; the occurrence of depression,1 point; the questionnaire for dyskinesia:the onset age ≤54,2 points; the occurrence of falling,3 points; daily levodopa dosage ＞ 600 mg,2 points.In all patients in this study,21.7％ (10/46) was asscssed to a total scorc of 0-1 which is associated with a low risk of motor fluctuation,8/16 had a score of 2 which is associated with intermediate risk and 8/9 got a score of 3-4 associated high risk; 10.2％ (5/49) had a score of 0-2,a low risk of dyskinesia,4/13 had a score of 3-4,a intermediate risk and 7/9 got a score of 5-7 which is associated with a high risk.Conclusions Age onset ≤54,the occurrence of swallowing difficulty,falling and depression,daily levodopa dosage ＞ 600 mg were considered to be the prognostic factors of motor complications in PD.The questionnaire may help to stratify PD patients into low-risk,medium-risk and high-risk groups for motor complications and the higher the score in the questionnaire is related to the higher risk of motor complications.

Objective To evaluate the olfactory function and its influence factors by using Sniffin ’ Sticks test, and to compare the quality of Parkinson ’s disease (PD) recognition between Sniffin’ Sticks and 16 kinds of odor identification in Sniffin ’ Sticks(SS-16) tests.Methods The Sniffin’Sticks test was used to assess the olfactory function of 68 PD patients and 76 healthy volunteers , and the relationship between smell and age, disease duration, Unified Parkinson’ s Disease Rating Scale score, Hoehn-Yahr (H-Y) rating, and cognitive function level (Montreal Cognitive Assessment) was analyzed.Results (1)The prevalence of olfactory dysfunction in PD group (83.3%) was significantly higher than that in control group (21.2%).The Sniffin’ Sticks test showed that the odor threshold score (6.6 ±3.2, P=0.000), odor discrimination score (6.6 ±3.3, P=0.000), 16 kinds of odor identification score (6.8 ±2.4, P=0.000) in PD group were significantly lower than those in control group.( 2 ) When comparing the PD cases and healthy controls in recognition , the sensitivity and the specificity of the Sniffin ’ Sticks test were 0.897 and 0.737, respectively, similar to the SS-16 test.However, the Sniffin’ Sticks test showed advantage compared with odor threshold and odor discrimination.( 3 ) The olfactory score in PD group was positively correlated with cognitive function (r=0.243, P=0.046), and was unrelated with age, gender, disease duration, and disease severity.The olfactory score in control group was negatively correlated with age (r=-0.270, P=0.018), but positively correlated with cognitive function (r=0.281, P=0.014).Conclusions There is a higher incidence of olfactory dysfunction in PD patients than in control group.Sniffin’ Sticks test is superior to SS-16 test in quantitative and qualitative analysis of olfactory function in PD patients.Two tests both have high sensitivity and specificity in the recognition of PD .

Sanjie Zhentong capsules were scanned by using a near infrared spectra probe with different drug mass fraction and the spectral information of capsule shells and contents in it were obtained. Then partial least squares (PLS) models were developed for the prediction of mass fraction of Fritillariae Thunbergii Bulbus and Resine draconis in Sanjie Zhentong capsules. The correlation coefficient (r9c)) and root mean standard error( RMSEC) of 0.949 5, 0.958 2 and 4.742 4, 4.135 7. The models obtained correlation coefficient (r(v)) of 0.919 2, 0.936 7 and root mean square error (RMSECV) of 6.158 9, 5.037 3 respectively in the training set. The paired T test analysis of statistics showed that there were no significant difference between predictive values and measure values. The established models reflected a strong prediction performance and can meet the needs of the production.
Capsules ; chemistry ; Drugs, Chinese Herbal ; chemistry ; Least-Squares Analysis ; Spectroscopy, Near-Infrared ; methods