OBJECTIVETo observe the impacts on the muscle strength in the patients of stroke-induced acroparalysis treated with Xunjingcuiqi needling technique.

METHODSOne hundred patients were randomized into a Xunjingcuiqi group and a routine acupuncture group, 50 cases in each group. In the routine acupuncture group, the routine acupuncture technique was adopted at the main acupoints, such as Shangxing (GV 23), Baihui (GV 20), Dicang (ST 4), Quchi (LI 11), Huantiao (GB 30) and Zusanli (ST 36), etc. In Xunjingcuiqi group, on the basis of the routine acupuncture technique, Xunjingcuiqi needling technique (pricking technique was quickly applied with the filiform needle along the running course of meridian to promote the conduction of meridian qi) was added. For the patients being hard to feel the needling sensation and with the muscle strength of 0 to 1 degree, Dongzhencuiqi technique was supplemented at shu-stream points of yang meridians (after qi arrival, the needling manipulation with limb movement was given to promote the conduction of meridian qi). The treatment was given once every day in the two groups. Ten treatments made one session. Three sessions of treatment were required. At the end of each session treatment, the muscle strength and clinical efficacy were assessed.

RESULTSIn the 1st, 2nd and 3rd sessions of treatment, 20, 24 and 36 cases achieved the 3 to 5 degrees muscle strength in Xunjingcuiqi group, respectively; and 6, 10 and 15 cases achieved the 3 to 5 degrees muscle strength in the routine acupuncture group. The differences were significant statistically in comparison of the two groups (P < 0.01, P < 0.05). The markably effective rates were 60.0% (30/50), 64.0% (32/50) and 70.0% (35/50) after the 1st, 2nd and 3rd sessions of treatment in Xunjingcuiqi group, respectively; and those were 38.0% (19/50), 44.0% (22/50) and 46.0% (23/50) in the routine acupuncture group, respectively. The differences were significant in the 1st and 3rd sessions of treatment between the two groups (both P < 0.05).

CONCLUSIONXunjingcuiqi needling technique combined with routine acupuncture achieves the apparent superior efficacy on acroparalysis induced by ischemic stroke as compared with the simple routine acupuncture. Xunjingcuiqi needling technique obviously improves muscle strength and shortens the duration of sickness.

Acupuncture Points ; Acupuncture Therapy ; instrumentation ; methods ; Aged ; Female ; Humans ; Male ; Middle Aged ; Paralysis ; etiology ; therapy ; Qi ; Stroke ; complications ; Treatment Outcome

OBJECTIVETo study the effect of Tiaoxin Recipe (TXR) on learning and memory related gene expression in hippocampus of senescence accelerated mice (SAM).

METHODSChanges of learning and memory related gene expression, including mineralocorticoid receptor (MR), presenile protein 1 and 2 (PS-1, PS-2), tau, APP, apoE and bcl-2 in hippocampus of SAM were determined by reverse transcription polymerase chain reaction (RT-PCR). The effect of TXR were tested. E2020 was used as the drug for control.

RESULTSCompared with those in the same aged mice, in the 5-month old SAM, levels of gene expression of MR, tau, PS-2 and APP were significantly higher, that of apo-E lower, levels of gene expression PS-1 and bcl-2 were unobviously changed; while in the 12-month old SAM, gene expression of MR and tau were higher, bcl-2 was lower and PS-1, PS-2, apoE and APP were also unobviously changed. Continuously orally taken TXR could correct the abnormality of MR, tau and apoE gene expression in hippocampus of 5-month SAM and that of MR and bcl-2 in 12-month SAM.

CONCLUSIONContinuously orally taken of TXR has the effect of regulating and correcting learning and memory related gene expression in hippocampus of 5-month and 12-month SAM.

Aging ; drug effects ; metabolism ; Animals ; Apolipoproteins E ; biosynthesis ; genetics ; Gene Expression ; Hippocampus ; metabolism ; Learning ; drug effects ; Membrane Proteins ; biosynthesis ; genetics ; Memory ; drug effects ; Mice ; Presenilin-1 ; Presenilin-2 ; tau Proteins ; biosynthesis ; genetics

Objective:To compare the Alzheimer's disease model in two species of mice by intracerebroventricular injection of β-amyloid peptide 25-35(Aβ_(25～35)).Method:The step down test and Morris water-maze were used to investigate the influence of the mice's learning and memorizing ability after intracerebroventricular injection of β-amyloid.Results:Normal Kunming mice and BALB/c mice had no significant difference in step down test, but the space cognitive ability of Kunming mice was better than that of BALB/c mice. The learning response in step-down test and Morris water-maze is no influence in male, female BALB/c mice and female Kunming mice. Marked differences were observed in male Kunming mice in acquisition, performance and reversal of a place learning response in a Morris water-maze.Conclusion:The male Kunming mice is much better than female Kunming mice and male, female BALB/c mice in preparing Alzheimer's disease animal model by intracerebroventricular injection of Aβ_(25～35), and is the most suitable animal in this experiment.

As an important neurotransmitter, adenosine displays its functions by acting on the adenosine receptors. Recent studies have shown that the distribution, expression and balance among subtypes of adenosine receptors are closely related with cognitive activities, and changes of adenosine receptors play key roles in neurodegenerative disorders including Alzheimer's disease. It has been pointed out that prolonged activation of adenosine receptors by high level adenosine may lead to the disturbance of balance among adenosine receptor subtypes. This imbalance mainly performed as increased expression of A2a receptor and decreased expression of A1 receptor, and enhancement of the excitatory signals mediated by A2a receptor and weakened inhibitory signals mediated by A1 receptor. Changes of these two subtypes of adenosine receptors may lead to a lot of disorders of neurological activities which developed into dysfunction of cognition to the end. These findings imply that the potential of maintaining the balance among adenosine receptors on the treatment of AD would facilitate both the revealing of the mechanism and the cure of AD.
Adenosine ; physiology ; Alzheimer Disease ; physiopathology ; Humans ; Neurotransmitter Agents ; physiology ; Receptors, Purinergic P1 ; classification ; physiology

Alzheimer' s disease (AD) is the most common form of dementia in the elderly. AD is an invariably fatal neurodegenerative disorder with no effective treatment. Senescence-accelerated mouse prone 8 (SAMP8) is a model for studying age-related cognitive impairments and also is a good model to study brain aging and one of mouse model of AD. The technique of cDNA microarray can monitor the expression levels of thousands of genes simultaneously and can be used to study AD with the character of multi-mechanism, multi-targets and multi-pathway. In order to disclose the mechanism of AD and find the drug targets of AD, cDNA microarray containing 3136 cDNAs amplified from the suppression subtracted cDNA library of hippocampus of SAMP8 and SAMR1 was prepared with 16 blocks and 14 x 14 pins, the housekeeping gene beta-actin and G3PDH as inner conference. The background of this microarray was low and unanimous, and dots divided evenly. The conditions of hybridization and washing were optimized during the hybridization of probe and target molecule. After the data of hybridization analysis, the differential expressed cDNAs were sequenced and analyzed by the bioinformatics, and some of genes were quantified by the real time RT-PCR and the reliability of this cDNA microarray were validated. This cDNA microarray may be the good means to select the differential expressed genes and disclose the molecular mechanism of SAMP8's brain aging and AD.
Aging ; genetics ; metabolism ; Alzheimer Disease ; genetics ; metabolism ; Animals ; Disease Models, Animal ; Gene Expression ; Gene Expression Profiling ; Hippocampus ; metabolism ; Male ; Mice ; Models, Animal ; Oligonucleotide Array Sequence Analysis ; Reverse Transcriptase Polymerase Chain Reaction

Objective To investigate the prognosis of patients with solitary large hepatocellular carcinoma (SLHCC) and with small hepatocellular carcinoma (SHCC),and analyze the risk factors affecting the prognosis of patients with SLHCC.Methods The retrospective case-control study was conducted.The clinicopathological data of 856 patients with hepatitis B virus (HBV)-related HCC who were admitted to the Eastern Hepatobiliary Surgery Hospital of the Second Military Medical University from January 2008 to December 2008 were collected.Of 856 patients,693 HCC patients with tumor diameter ≤5 cm were allocated into the SHCC group and 163 HCC patients with tumor diameter ＞ 5 cm and with solitary,expansive growth and complete capsule tumors were allocated into the SLHCC group.Patients underwent preoperative antiviral therapy,laboratory and imaging examinations,and then surgical planning was determined based on the preoperative results.Observation indicators:(1) comparisons of clinicopathological features between the 2 groups:sex,age,Child-Pugh grade,HBeAg,serum level of HBV-DNA,platelet (PLT),albumin (Alb),total bilirubin (TBil),alpha-fetoprotein (AFP),tumor diameter,microvascular invasion,Edmondson-Steiner grade and liver cirrhosis;(2) treatment situations between the 2 groups:surgical procedures,operation time,volume of intraoperative blood loss,number of patients with blood transfusion and time of hepatic inflow occlusion;(3) survival analysis between the 2 groups;(4) prognostic analysis of patients with SLHCC.Follow-up using telephone interview and outpatient examination was performed once every 3 months within 2 years postoperatively and once every 6 months after 2 years postoperatively up to June 23,2014.Follow-up included tumor marker,liver function,serum level of HBV-DNA and abdominal B-ultrasound examination.The patients received reexamination of computed tomography (CT) or magnetic resonance imaging (MRI) once every 6 months or when there was suspicion of tumor recurrence or metastasis.Tumor recurrence or metastasis was confirmed through typical HCC imaging findings of CT and MRI,and PET/CT examination was conducted if necessary.Tumor-free survival time was from operation time to time of tumor recurrence,and overall survival time was from operation time to death or the last follow-up.Measurement data with normal distribution were represented as-x±s,and continuous variables were analyzed by the t test or Mann-Whitney U test.Measurement data with skewed distribution were described as M (range).Categorical variables were represented as count (percentage) and analyzed by the chi-square test or calibration chi-square test.The survival curve and survival rate were respectively drawn and calculated by the Kaplan-Meier method and Log-rank test.COX regression model was used for prognostic analysis.Results (1) Comparisons of clinicopathological features between the 2 groups:number of patients with PLT＜ 100× 109/L,with positive microvascular invasion and with liver cirrhosis and tumor diameter were 197,133,447,(3.1±1.1)cm in the SHCC group and 28,53,79,(8.9±3.3) cm in the SLHCC group,respectively,with significant differences between the 2 groups (x2=28.618,t =37.286,x2 =213.773,214.325,P ＜ 0.05).(2) Treatment situations between the 2 groups:all the 856 patients underwent hepatectomy,including 326 with hepatic segments of resection ≥ 3 and 530 with hepatic segments of resection ＜ 3.Operation time,volume of intraoperative blood loss,number of patients with intraoperative blood transfusion and with time of hepatic inflow occlusion ＞ 20 minutes were 90 minutes (range,60-200 minutes),200 mL (range,20-5 200 mL),47,125 in the SHCC group and 110 minutes (range,60-230 min),300 mL (range,50-3 200 mL),31,58 in the SLHCC group,respectively.(3) Survival analysis between the 2 groups:all the 856 patients were followed up for 32.5 months (range,1.O-72.3 months).The median survival time,median tumor-free survival time,1-,3-,5-year overall survival rates and 1-,3-,5-year tumor-free survival rates were 56.2 months (range,1.6-75.8 months),39.5 months(range,1.0-75.0 months),90％,71％,58％,70％,48％,38％ in the SHCC and 50.3 months (range,1.1-76.0 months),30.7 months (range,1.0-72.0 months),87％,59％,47％,65％,46％,33％ in the SLHCC group,respectively,with no significant difference in tumor-free survival between the 2 groups (x2=0.514,P＞0.05) and with a significant difference in overall survival between the 2 groups (x2=10.067,P＜0.05).Stratified analysis:there were 117 SLHCC patients with 5 cm ＜ tumor diameter ＜ 10 cm and 46 SLHCC patients with tumor diameter ＞ 10 cm.The 1-,3-,5-year overall survival rates and 1-,3-,5-year tumor-free survival rates were 91％,65％,53％,70％,48％,35％ in 117 SLHCC patients with 5 cm ＜ tumor diameter ＜ 10 cm,respectively,with no significant difference compared with SHCC group (x2=1.832,0.042,P＞0.05).The 1-,3-,5-year overall survival rates and 1-,3-,5-year tumor-free survival rates were 78％,46％,31％,49％,39％,30％ in 46 SLHCC patients with tumor diameter ＞ 10 cm,respectively,with significant differences compared with SHCC group (x2=21.136,4.097,P＜0.05).(4) Prognostic analysis of patients with SLHCC:results of univariate analysis showed that serum level of HBV-DNA,tumor diameter and microvascular invasion were risk factors affecting postoperative 5-year tumor-free survival rate of SLHCC patients (x2 =5.193,3.377,5.509,P＜0.05);sex,serum level of HBV-DNA,tumor diameter and microvascular invasion were risk factors affecting postoperative 5-year overall survival rate of SLHCC patients (x2=4.546,18.053,7.780,10.569,P＜0.05).Results of multivariate analysis showed that serum level of HBV-DNA ≥ 104 U/mL,tumor diameter ＞ 10 cm and positive microvascular invasion were independent risk factors affecting postoperative 5-year tumor-free survival rate of SLHCC patients [HR =2.77,1.85,1.86,95％ confidence interval (CI):1.74-4.40,1.16-2.94,1.17-2.96,P＜ 0.05] and affecting postoperative 5-year overall survival rate of SLHCC patients (HR=2.73,1.98,1.69,95％CI:1.72-4.33,1.23-3.17,1.04-2.72,P＜0.05).Conclusions There are similar prognosis between SLHCC patients with 5 cm ＜ tumor diameter ＜ 10 cm and SHCC patients,however,prognosis of SLHCC patients with tumor diameter ＞ 10 cm is worse than that of SHCC patients.Serum level of HBV-DNA ≥ 104 U/mL,tumor diameter ＞ 10 cm and positive microvascular invasion are independent risk factors affecting prognosis of SLHCC patients.

Objective To prepare a glycerol reference material.Methods The material was prepared and characterized according to the primary standard substance technological specification(JJG 1006- 1994).Glycerol was dissolved in water containing 0.5% sodium azide and dispersed to glass ampules.The homogeneity and stability of this material were tested with an HPLC method.Glycerol concentration was determined by a titration method as specified in the Pharmacopoeia of China.Results The three time measuring result of glycerol reference material was 1.297 5?0.014 3,1.302 0?0.008 9,1.313 7? 0.007 8,respectively.Statistical analysis showed that this material was homogeneous (F=1.462,P=0.166) .It was stable for at least 4 years at 4℃.The assigned reference value was 0.103 6 g/g and the expanded uncertainty was 0.000 4 g/g.Conclusions This material meet the technical requirements of national primary standard reference material.It is approved as the Certified Reference Material (GBW 09149) by General Administration of Quality Supervision,Inspection,Quarantine of the People's Republic of China in May,2006.

By non-steady method,the effective diffusivity of ferrous sulphate within alginate calcium gel entrapped without bacteria was measured.Meanwhile the oxidation ability of entrapped bacteria was analyzed.Experimental results showed that the effective diffusion coefficient of ferrous sulphate decreased with the increase of alginate concentration,the optimum alginate concentration is 2%(W/V).The effect of calcium chloride on the effective diffusivity was neglectable.The incubation of ferrooxidans would pass through 10 hours,and the diffusion coefficient within gel entrapped Thiobacillus ferrooxidans cells was less remarkably than that of ferrous sulphate without entrapped cells.For the entrapped cells,the absolute oxidation time was shortest and the rate change was fastest with the initial Fe concentration 5g/L.The absolute oxidation time was same when the initial Fe concentration was 8g/L and 10g/L.

In order to increase the production of insecticidal crystal proteins Cry1 and Cry2, firstly, Plack-ett-Burman design was applied to evaluate the effectiveness of the related nutrition factors; it was found that the soybean powder and MnSO4?H2O were significant factors for Cry1 production, but the yield of Cry2 wasn’t effected remarkably in such medium. Then the steepest ascent experiment was adopted to approach the optimal region of the medium composition. Lastly, the optimal concentration of the soybean powder and MnSO4?H2O was 11.5 and 0.02 g/L, obtained by response surface methodology (RSM). The final yields of Cry1 and Cry2 was 0.32 mg/mL and 0.11 mg/mL, increasing twice more than that in the medium optimized before. The median lethal concentration (LC50) of optimal medium was 1.09 ?L/mL. The toxicity to Heli-coverpa armigera was significantly enhanced than the old one.

OBJECTIVE To establish an in vitro cell model based on patient-specific human neural stem cells to study the pathomechanism of sporadic AD as well as screen candidate drugs.METHODS The peripheral blood cells from sporadic AD patients and cognitive normal controls were repro-grammed into inducedpluripotent stem cells(iPSCs),which were further induced into neural stem cells and neurons. The cell growth curve during the differentiation process was recorded by the IncuCyte ZOOM, and neural stem cells and neurons were identified by immunofluorescence. The apoptosis of neural stem cells and neurons was detected by Click-iT?Plus TUNEL Assay. RESULTS Neural stem cells derived from AD patients and cognitive normal controls can express neural stem cell markers Nes-tin,Sox1,Sox2 and Ki67.TUNEL assay results showed that the number of TUNEL-positive cells in neu-ral stem cells derived from AD patients was significantly higher than that of cognitive normal controls (P<0.01). When neural stem cells were differentiated into neurons, the percentage of MAP2 positive cells in the neural stem cell-derived culture dish of AD patients was significantly higher than the cogni-tive normal controls at day 16 of neuronal differentiation (P<0.01); the TUNEL assay showed that the number of TUNEL-positive cells in AD-derived neurons was significantly greater than that in cognitive normal controls (P<0.01) at day 16 of neuronal differentiation. CONCLUSION Our study revealed that AD-iPSC-derived neural stem cells exhibit premature neuronal differentiation and increased neural apoptosis,which might be relevant to the neuronal loss of AD,thus may provide valuable new tools to screen candidate drugs for the disease and to discover the mechanisms underlying AD pathogenesis.