Objective To observe the therapeutic efficacy of Tiao Shen Tong Luo (regulating mind and unblocking collaterals) needling plus hyperbaric oxygen in treating leukoaraiosis (LA) coupled with mild cognitive impairment (MCI). Method Totally 112 patients with LA and MCI were randomized into a treatment group and a control group. In addition to the basic treatment, the treatment group was intervened by Tiao Shen Tong Luo needling plus hyperbaric oxygen, while the control group was given hyperbaric oxygen. The scales and event-related potential (P300) were evaluated and compared before and after the intervention. Result The scales scores and P300 were significantly changed in both groups after the treatment, and the therapeutic efficacy was higher in the treatment group than in the control group (P<0.05). Conclusion In addition to the basic treatment, Tiao Shen Tong Luo needling plus hyperbaric oxygen can produce a significant efficacy in treating LA couple with MCI.

Objective To investigate the changes of leptin level in patients with intracerebral hemorrhage and the relationship between leptin level and insulin resistance(IR). Methods 57 patients with hypertensive cerebral hemorrhage (group A ), 50 patients with hypertension (group B), 41 healthy people (group C) were selected to determine the value of fasting blood-glucose ( FBG), fasting blood-insulin( FBI), leptin levels as well as insulin sensitivity index (ISI). Results The level of FNS and leptin was significantly higher (P ＜ 0.05 ) while the level of ISI was lower (P ＜ 0.05 ) in group A than those in group B and C. Correlation analysis showed that leptin level in HCH patients was negatively correlated with ISI, and positively correlated with FNS. Conclusion The results indicate that higher level of leptin and IR may play an important role in the occurrence of hypertension complicated cerebral hemorrhage.

Objective To observe the clinical effect of abdominal Tuina and acupuncture in treating Swiss generalized anxiety disorder (GAD). Methods 80 patients of GAD caused by liver depression-qi stagnation were randomly recruited into a treatment group and a control group, 40 patients in each. The treatment group was treated with abdominal tuina & acupuncture; while the control group was treated with acupuncture exclusively. Clinical effects and hamilton anxiety scale (HAMA) score were observed in both groups before and after the treatment. Results ①There was significant difference in total efficacy between the two groups CP<0.05), and the treatment group was better than the control group. ②The HAMA score of the two groups after the treatment was lower than that before the treatment, and the difference was significant (P< 0.05). There was also significant difference of HAMA score between the two groups after the treatment (/><0.05), with the treatment group showing better results than the control group. Conclusion Abdominal tuina combined with acupuncture has sound effects in treating Swiss GAD due to liver depression-qi stagnation.

Object To investigate the effects of Heart-benefitting Recipe (HBR) on the histopathological changes of Alzheimer's disease (AD) rats after unilateral amyloid-?_ 25-35 protein (A?_ 25-35 ) injection into the amygdala. Methods The experimental rat models with dementia,spatial learning,and memory impairment were induced by unilateral amyloid ?_ 25-35 protein (A?_ 25-35 ) injection into the amygdala of rats. The spatial learning and memory ability and the function of cholinergic system were observed by the means of Morris water maze and radioligand binding assay. The protein expression of A?_ 1-40 ,AT_8 was estimated with immunohistochemistry. The expression of mRNA related to APP was observed by RT-PCR. Results HBR significantly alleviated the spatial learning and memory impairment induced by A?_ 25-35 and remarkably increased the activity of ChAT and Rt of M-receptor binding sites of these models. The expressions of A?_ 1-40 protein and APP mRNA in cortex and hippocampus of AD rats were decreased remarkably by HBR. The results of immunohistochemistry showed that the expression of hyperphosphorilated tau AT_ 8 in cortex and hippocampus of AD rats was inhibited significantly by HBR. Conclusion HBR can effectively improve the spatial learning and memory impairment,decrease the deposition of A?_ 1-40 ,and alleviate the hyperphos-phorylation of tau of AD rats induced by A?_ 25-35 .

OBJECTIVE:To establish a method for the content determination of verbascoside in Naosaitong honeyed pill. METHODS:HPLC was performed on the column of Zorbax ODX with mobile phase of acetonitrile-0.1% phosphoric acid(16∶84, V/V)at a flow rate of 1.0 ml/min,the detection wavelength was 334 nm,column temperature was 30 ℃,and the injection volume was 10 μl. RESULTS:The linear range of verbascoside was 0.200-1.000 μg(r=0.999 8);RSDs of precision,stability and repro-ducibility tests were lower than 1%;recovery was 95.16%-99.78%(RSD=1.80,n=6). CONCLUSIONS:The method is simple and accurate,and suitable for the content determination of verbascoside in Naosaitong honeyed pill.

Objective To explore the application of diffusion tensor imaging(DTI) in neonates with hypoxic ischemic encephalopathy(HIE).Methods 37 cases of full-term neonates diagnosed as HIE by pediatricians and 12 normal neonates were selected.All subjects underwent MRI and DTI at 3.0 Tesla.The correlation of FA value in posterior limb of internal capsule and neonatal NBNA score was examined.Results The FA values in posterior limb of internal capsule, splenium had significant difference between the HIE group (mild group, moderate group, severe group) and control group (P<0.05);in central white matter of the frontal lobe, thalamus had significant difference between HIE severe group and control group (P<0.05);in nucleus of the bean had no statistic difference between the HIE group (mild group, moderate group,severe group) and control group (P>0.05).The FA in posterior limb of internal capsule and NBNA score(12-14 d;26-28 d)showed significant correlation (r=0.96,P<0.05),as well as the sequelae shape (r=0.87,P<0.05).Conclusion DTI is promising for early diagnosis of anisotropic index changes to evaluate HIE brain damage and prognosis.

ObjectiveTo investigate the association of phosphorylation of mammalian target protein of rapamycin (pmTOR) expression with glioma malignancy grades,and the correlation of pmTOR expression with Survivin and Ki-67,which represent tumor cell anti-apoptosis ability and reproductive activity.MethodsImmunohistochemistry EliVision method was employed to detect the expression of pmTOR,Survivin and Ki-67 in paraffin tissues from 87 patients with glioma (grade Ⅰ - Ⅱ 27 cases,grade Ⅲ24 cases and grade Ⅳ 36 cases).The association between positive expression rate,level of pmTOR and malignancy grades,and the correlation of its expression level with Survivin and Ki-67 were further evaluated.Results There was no significant difference in the positive expression rate of pmTOR among grade Ⅰ - Ⅱ(77.8％,21/27),grade Ⅲ(75.0％,18/24) and grade Ⅳ (72.2％,23/36) (P ＞ 0.05).However,the significant association between pmTOR expression level and malignancy grades was observed.The expression from 87 patients with glioma was significantly positively correlated with Survivin and Ki-67 expression level (r =0.858,P ＜ 0.01 ;r =0.708,P ＜ 0.01 ).ConclusionsThe expression level of pmTOR is associated with malignancy grades,tumor cell anti-apoptosis ability and reproductive activity.pmTOR may be served as a useful marker for predicting the biological behavior of glioma and a useful target for gene therapy.

Objective To construct recombinant expression vector TAT-SOX2 and express the fusion protein in E.coli BL21.Methods SOX2 fragment were amplified by PCR.The product was digested with restriction enzymes then was inserted into PET-28b-TAT-V2 which contains protein transduction domain of TAT.The recombinant vector was transformed into E.coli BL and induced with IPTG.The expressed fusion protein was analyzed by using SDS-PAGE method.The highly expressed product was purified by affinity chromatography.Western blotting was used to identify the specificity of the fusion protein.The immunofluorescence was used to detect the efficiency of fusion protein transduced to HSF cells.Results The recombinant expression vector TAT-SOX2 was correctly constructed and fusion protein TAT-SOX2 was successfully expressed in prokaryotic cells.Western blotting showed the specificity of the fusion protein.The immunofluorescence prompt fusion protein was transduced into HSF cells quickly.Conclusion This study provides the material basis for the further induced pluripotent stem cells by protein transduction.

Objective To construct recombinant expression vector TAT-KLF4 and express the Fusion Protein in E.coli BL21.Methods The open reading frame(ORF) of KLF4 gene was amplified by PCR.The PCR product was digested with restriction enzymes then was inserted into PET-28b-TAT-V2 which contained protein transduct domain of TAT.The recombinant vector was transfected into E.coli BL21 and the expression of the TAT-KLF4 fusion protein was induced with IPTG.The fusion protein was analyzed by using SDS-PAGE and purified by the affinity chromatography.Western Blot was used to identify the specificity of the fusion protein.The immunofluorescence was used to detection the efficiency of fusion protein transducted to HSF cells.Results The recombinant expression vector TAT-KLF4 was correctly constructed and fusion Proteion TAT-KLF4 was successfully expressed in prokaryotic cells.Western blotting showed the specificity of the fusion protein.The immunofluorescence prompt fusion protein transduced into HSF cells quickly.Conclusion This study provides the material basis for further induction of the induced pluripotent stem cells by protein transduction.

Objective To analyze the diagnostic value of parameters derived from intravoxel incoherent motion diffusion-weighted imaging (IVIM-DWI) and dynamic contrast-enhanced MRI (DCE-MRI) in cervical cancer and to explore the correlation of parameters between DCE-MRI and IVIM-DWI.Methods MRI data of 30 patients with cervical cancer confirmed by surgical pathology and 26 patients with normal cervix confirmed by clinical or MRI examination were analyzed retrospectively.All patients underwent IVIM-DWI and DCE-MRI.The IVIM-DWI was performed to calculate the values of ADC,D,D* and f.The values of Ktrans,Kep and Ve were measured on DCE-MRI.All parameters between cervical cancer and normal cervix were compared by two independent sample t-test.Diagnostic efficacy of these parameters in cervical cancer group was evaluated by using area under curve(AUC).The correlation of these parameters between IVIM-DWI and DCE-MRI was analyzed by Pearson correlation.Results The D*,f,Ktrans,Kep and Ve in cervical cancer were significantly higher than those in normal cervix.The ADC and D in cervical cancer were significantly lower than those in normal cervix.The differences of these parameters between the two groups were statistically significant (P ＜0.05).The AUC of Ktrans was the maximum (0.958),with diagnostic sensitivity of 86.7％ and specificity of 100％.F and K were positively correlated (r=0.446,P＜0.05).F and Kep were positively correlated (r=0.400,P＜0.05).Conclusion IVIM-DWI and DCE-MRI can improve the diagnostic value of cervical cancer.DCE-MRI has better diagnostic performance than IVIM-DWI.Significantly positive correlations were found between perfusion paramenters from IVIM-DWI and quantitative perfusion paramenters from DCE-MRI.