BACKGROUND: Effective proliferation in vitro of neural stem cells and directional induction and differentiation of dopaminergic neurons play a key role in neural stem cell transplantation for treatment of degeneration of the nervous system and repair of injured nervous system. OBJECTIVE: To explore the differentiation of neural stem cells from anterior subventricular zone into dopaminergic neurons in astrocyte-conditioned medium. DESIGN, TIME AND SETTING: The in vitro controlled cytology experiment was performed at the Experimental Center of Xinqiao Hospital of Third Military Medical University of Chinese PLA from December 2006 to August 2007. MATERIALS: A total of 15 neonatal KM mice aged 2 days and 40 pregnant 16 days Wistar rats were provided by the Experiment Animal Center of Third Military Medical University of Chinese PLA. METHODS: Astrocytcs of KM mice were isolated and purified by a standard shaking method and differential adhesion. Astrocyte-conditioned medium was collected when astrocytcs at passage 3 were cultured for 5 days maintained at –20 ℃ for use. Neural stem cells from the anterior subventricular zone of Wistar rats were isolated and cultured in serum-free DMEM/F12, supplemented with B27 and basic fibroblast growth factor for primary culture. Neural stem cells at passage 3 were collected and incubated at 0.5?108/L. Cells in the control group were incubated in the DMEM/F12 medium containing fetal bovine serum of 0.1 volume fraction. Cells in the experimental group were incubated in the astrocyte-conditioned medium. MAIN OUTCOME MEASURES: Neural stem cells in the anterior subventricular zone were determined by the immunocytochemical technique. Positive rate of the neural stem cell differentiation into dopaminergic neurons in the anterior subventricular zone was detected by the flow cytometry. RESULTS: Cells in neurospheres expressed nestin and differentiated into neuro-specific enolase and glial fibrillary acidic protein. Tyroxine hydroxylase positive cells were found in the experimental group, showing round or ellipse. Tyroxine hydroxylase was seen in cytoplasm and cluster-shape processes. Tyroxine hydroxylase positive cells in control group were different from that in the experimental group in amount and appearance. Positive cell rate of tyroxine hydroxylase was significantly higher in the experimental group than in the control group (t =35.296, P

Objective To investigate the influencing factors of progressive hemorrhagic injury (PHI) after traumatic brain injury. Methods The medical records of 127 patients with traumatic brain injury (n=49 in PHI group and n=78 in non-PHI group) were reviewed. The relationship between PHI and influencing factors including sex, age, Glasgow coma scale, time from injury to first CT, traumatic subaraehnoid hemorrhage (tSAH), prothrombin time(PT),activated partial thromboplastin time(APTT) was analyzed. Results The time for first CT was(1.39± 1.27) h in PHI group and (2.91±1.85) h in non-PHI group (t=2.14, P<0.05). 35 cases of PHI group developed tSAH and 37 of non-PHI group developed tSAH (χ2=7.06, P<0.05). Multifactor Logistic regression analysis showed that the time for first brain CT after injury and the patients accompanied with tSAH were associated with PHI after traumatic brain injury (OR=0.558,95 % CI 0.329-0.946, OR=13.000,95 % CI 1.187-142.354, P<0.05 for each). Conclusions Time from injury to first CT and tSAH can be prognostic factors for PHI.

Objective Dendritic cell-associatedC-type lectin-1 ( Dectin-1) is one of the most important receptors in antifungal innate immune response.This study was to construct a recombinant adenovirus vector expressing themurine Dectin-1gene and acquire a high-concentration adenovirus by amplification and purification. Methods The PCR amplification product CLEC7A-pIRES2-EGFP was cloned into the intermediate vector pDONR221, and then recom-bined with the backbone vector pAD/CMV/V5-DEST to produce a re-combinant plasmid pAD-CLEC7A-pIRE2S -EGFP.The recombinant plasmid was linearized with Pac I and transfected into human embryon-ic kidney ( HEK293) cells to produce recombinant adenovirus pAD-CLEC7Ap-IRES 2-EGFP. The adenovirus was propagated in the HEK293 cells and purified by filtering through the cellulose acetate membrane and concentrating column.Fluorescence microscopy and re-al-time PCR were used to determine the expression of the Dectin-1 gene. Results PCR identification, enzyme digestion, and sequen-cing results manifested theDectin-1 gene in the vector, with the final adenovirus titer of 5×1011 IU/mL.Fluorescence microscopy revealed green fluorescence and real-time PCR assay confirmed that the expression of Dectin-1 was improved by 8677.25 times. Conclusion A relatively high-titer adenovirus expressing Dectin-1 was acquired,which may help to further study the high expression of Dectin-1 in anti-fungal innate immunity in vitro and in vivo.

Using Dachengqi Tang (DCQT) as a model, high performance liquid chromatography (HPLC) fingerprints were applied to optimize machine extracting process with the Box–Behnken experimental design. HPLC fingerprints were carried out to investigate the chemical ingredients of DCQT; synthetic weighing method based on analytic hierarchy process (AHP) and criteria importance through intercriteria correlation (CRITIC) was performed to calculate synthetic scores of fingerprints; using the mark ingredients contents and synthetic scores as indicators, the Box–Behnken design was carried out to optimize the process parameters of machine decocting process under high pressure for DCQT. Results of optimal process showed that the herb materials were soaked for 45 min and extracted with 9 folds volume of water in the decocting machine under the temperature of 140 1C till the pressure arrived at 0.25 MPa;then hot decoction was excreted to soak Dahuang and Mangxiao for 5 min. Finally, obtained solutions were mixed, filtrated and packed. It concluded that HPLC fingerprints combined with the Box–Behnken experimental design could be used to optimize extracting process of traditional Chinese medicine (TCM).

Objective To identify the imaging characteristics of calcifying pseudoneoplasms of the neuraxis (CAPNON) and do literature review.Methods Five patients of pathologically-proved CAPNON underwent preoperative MR examination,among which 4 underwent CT scan,2 underwent DSA examination and 1 underwent SPECT. All imaging data were retrospectively analyzed with the emphasis on imaging characteristics.Results Five patients of CAPNON with the diameter of 1.5 to 5.0 cm were found in five patients ( Male 4 ; Female 1 ; age 25 to 60 years old ).Three lesions were located in the skull base,one was located in the cervical spine and one in the foramen magnum and upper cervical segment. All patients underwent MRI examination and 4 of them also took CT scanning.On plain CT,all lesions showed obvious calcification.On T1WI all masses showed hypointensity,and on T2WI 4 of the lesions showed iso- or hypointensity and 1 heterogeneous signal intensity. On contrast-enhanced MR images, peripheral enhancement was demonstrated in 3 lesions,homogeneous enhancement was found in case and one lesion showed no enhancement. The pathologic analysis indicated that inside the lesions were abundant calcification,fibroepithelial tissue and mucoid matrix and no edema was detected around the lesions.Conclusions CAPNON displayed the predilection to male adults and the neuraxis was the predilection site.Calcification on CT images,hypointensity on MR images and peripheral enhancement will be helpful for the diagnosis of CAPNON,but the final confirmation still needs the pathologic results.

Objective The objective of the present study was to establish a template for the self assessment of hearing aids outcomes according to the evaluation of a large group of hearing aid users .Methods In total ,1 724 subjects participated in the study .The Chinese version of International Outcome Inventory for Hearing Aids (IOI-HA) was used as the evaluation tool .IOI -HA is a seven -item questionnaire ,each item is designed to assess a specific outcome domain .Each item was designed with a five -point rating scale ,a higher rating indicates better outcome .According to the subjects'self reported hearing difficulty when they were not wearing hearing aids (unaid-ed) ,the subjects were divided into two groups :self reported hearing difficulty to be no ,mild ,moderate group ,and self reported hearing difficulty to be moderately severe or severe group ,respectively .Templates for each group were established according to the analysis of the item ratings .Results One thousand two hundred and forty -seven sub-jects accept the investigation ,with a response rate of 72 .3% ,and 1203 completed responses were included in the fi-nal analysis .The Chinese version of IOI-HA scores showed a skewed distribution ,with a mean score ranging from 3 .52 to 4 .19 .The total IOI-HA scores ranged from 9 to 35 ,and the mean total score was 26 .30 .Results showed that for the self reported hearing difficulty (unaided) to be none ,mild or moderate group ,the mean scores for the i-tem of daily use ,benefit ,residual activity limitation ,satisfaction ,residual participation restrictions ,impact on others ,and quality of life were 4 .14 ,3 .67 ,3 .76 ,3 .58 ,3 .56 ,3 .93 and 3 .69 ,respectively .The corresponding mean scores for the self reported hearing difficulty (unaided) to be moderately severe or severe group were 4 .19 ,3 .63 ,3 .64 ,3 . 65 ,3 .52 ,3 .89 ,and 3 .79 ,respectively .A template for the Chinese IOI -HA was established according to the above data .Conclusion The template for the Chinese version of IOI -HA could be served as an effective tool to measure the general effectiveness of the hearing aid outcomes and hearing aid fitting in China ,which would also facilitate the international outcome comparison cross culture .

Objective To investigate the effects of social support intervention on anxiety and depression of pulmonary hypertension(PH)patients,then provide a scientific basis for nursing of patients with pulmonary hypertension.Methods The general condition of PH questionnaire,self-rating anxiety scale (SAS),self-rating depression scale(SDS),social support rating scale(SSRS)were distributed to 131 patients with PH.Then make statistical analysis of patients'anxiety,depression and social support conditions.Results The score of anxiety and depression psychological conditions of patients with pulmonary hypertension was significantly higher than normal population,the difference wag statistically significant.Among 131 patients,16 patients with anxiety,accounting for 12.21% ;21 cases of patients with depression,accounting for 16.03% .28 patients at a high level of social support,92 patients at a medium level of social support,11 patients at a low level of social support,a total of 91.60% of the patients in the middle and higher levels of social support.Anxiety and depression scores had significant negative correlation with social support,objective support points and subjective support points.The anxiety,depression difference among different types of pulmonary hypertension was statistically significant.The difference of anxiety and depression scores between patients with idiopathic pulmonary arterial hypertension and patients with pulmonary hyper tension caused by congenital heart disease were significant.The depression scores between pulmonary hy pertension caused by pulmonary veno-occlusive disease and congenital heart disease were significantly different.Conclusions When nurses care pulmonary hypertension patients.those with different types of PH should be given targeted social support.Attention should be paid to transfer the source of social support to help them adopt a positive attitude to face the diseabe,then improve the treatment and care compliance of patients.

This paper summarizes the key points of safety management for 15 patients with severe pulmonary arterial hypertension treated by aerosolized iloprost. All patients achieved significant improvements and none of them suffered any severe side effect. Complete safety management during the therapeutic procedure improved the patients' treatment confidence and compliance,and thereafter strengthened the efficacy of treatment.

Objective To investigate the prevalence of mutation in the locus 306 of embB gene in multi-drug resistant (MDR) Mycobacterium tuberculosis (TB) and evaluate the prospects for using it as a molecular marker to detect MDR-TB.Methods The 291 strains enrolled in this study were from the reference laboratory of Shanghai municipal centers for disease control and prevention, all of which had been tested for drug susceptibility.Mutation in embB 306 was screened both by amplification refractory mutation system (ARMS) and DNA sequencing.The mutation frequencies of embB 306 in the sample groups varied in drug resistance were statistically analyzed.Results 38(51.4% ) of the 74 MDR-TB were embB 306-mutant (X2 =93.8,P＜0.01).Of the 24 TB resistant to at least two drugs but not MDR, 9(37.5% ) were embB 306 mutant (X2 =60.1 ,P＜0.01 ).But only two(4.9% ) embB 306-mutant strains were found in 41 strains resistant to only one drug (X2 =6.8,P=0.0093).None embB 306-mutant strains were found in 152 pansensitive strains.The specificity of using embB 306 as a molecular marker for detecting multi-drug resistant TB was 94.9% (206/217).Conclusions As a molecular marker for screening drug resistant TB,especially MDR-TB, the gene locus embB 306 shows a relatively high sensitivity and specificity, promising a sound future for its application in clinics to realize fast screening of patients infected with MDR-TB and to provide evidence for appropriate medication.

Objective:This study was performed using preclinical transplanted animal experiments to analyce the effects and mechanisms of third-generation EGFR-TKIs combined with anti-angiogenic therapy, thereby providing theoretical basis for further clinical trials. Methods:Researchers constructed the transplant BALB/C nude mice models with H1975 lung adenocarcinoma cell line (EGFR T790M) and divided the mice into four groups and treated them with osimertinib (2.5 or 5 mg/kg/day, gavage) alone or plus bevacizumab (5 mg/kg/twice weekly, i.p.) when the tumors reached approximately 0.4-0.6 cm3 in volume. The tumor growth curve of tumor volume was drawn according to the time in every group. After 2 weeks of treatment, the mice were killed and the tumors were processed for immunohistochemical staining and Western blot analysis. Immunostaining was performed to detect:HIF-1α, VEGF, and microvessel density (MVD) by using SP method on paraffin sections. Western blot analysis was used to analyze the protein expression levels of EGFR, AKT, and ERK signal transduction pathways. Results:After 2 weeks of treatment in high-and low-dose osimertinib alone, tumor volume in the high-dose group was significantly less than in low-dose osimertinib-alone group (P<0.05). VEGF, HIF-1αexpression, and MVD were significantly low in the high-dose osimertinib-alone group (P<0.05). Increasing doses of osimertinib induced dose-dependent weakening of the p-EGFR, p-AKT, and p-ERK expression levels (P<0.05). In the low-dose osimertinib-plus-bevacizumab group and low-dose osimertinib-alone group, no significant difference in tumor volume and the above factors was observed. In the low-dose osimertinib-plus-bevacizumab group and high-dose osimertinib-alone group, tumor volumes did not exhibit significant difference (P=0.178). Moreover, VEGF, HIF-1αexpression, and MVD exhibited no significant difference. No significant difference in the p-EGFR, p-AKT, and p-ERK expression levels was found between high-dose osimertinib-alone group and low-dose osimertinib-plus-bevacizumab group (P>0.05). In the high-dose osimertinib-plus-bevacizumab group, tumor growth was not significantly greater than that in the high-dose osimertinib-alone group (P=0.642). No significant difference was observed in the above factors.In the high-and low-dose osimertinib-plus-bevacizumab groups, tumor volume and the above factors did not exhibit significant differences (P>0.05). Conclusion:Osimertinib has obvious antitumor effects in EGFR-mutant lung adenocarcinoma with T790M mutation cell xenografts. Bevacizumab has a synergetic inhibitory effect with osimertinib against EGFR-mutant lung adenocarcinoma with T790M mutation cell xenografts. Bevacizumab enhanced the antitumor effects of osimertinib by reducing VEGF expression and the microvascular density of the tumor, thereby improving the tumor microenvironment. Bevacizumab can enhance the effect of osimertinib by suppressing EGFR, ERK, and AKT phosphorylation, thereby synergistically inhibiting EGFR activation and downstream signaling.