Objective To establish a mouse model of prostate cancer expressing human PSCA for the development of new anti-tumor drugs or vaccines. Methods The total RNA of DU145 cells,a human prostate cancer cell line,was isolated by using TRIzol reagent according to the (RT-PCR),the first-strand cDNA was synthesized using the SuperScript First-Strand synthesis system. The human PSCA gene was amplified with the primers and cloned into the plasmid pcDNA3.1 to generate pcDNA-PSCA. DNA sequencing was used to confirm the constructs. The mouse prostate tumor cell line RM-1 cells,syngeneic to C57BL/6,were transfected with pcDNA-PSCA plasmids followed by selection using G418. RT-PCR analysis was performed to examine the validity of the constructs. Expression of PSCA on the cell surface was determined by staining with anti-PSCA antibody,and the anti-PSCA antibody was detected using an FITC-conjugated goat anti-rabbit IgG antibody,and analyzed by flow cytometry. 4-6-week-old male C57BL/6 mice purchased from the Laboratory Animals Center were inoculated with different amounts of RM-PSCA cells to search for suitable cell population which can form tumor in mouse,and the mice were monitored twice a week. The growth and the survival time of mice were measured,respectively. The tumor volume was measured by vernier caliper according to the formula:V=0.5a×b~2,where a and b are the long and short diameters of the tumor,respectively. Results The plasmid pcDNA-PSCA was successfully constructed and the PSCA was successfully expressed in RM-PSCA 7~# and RM-PSCA 28~# cells by RT-PCR and confirmed by flow cytometry. 1×10~5 RM-PSCA cells were sufficient to get tumor growth in 100% of inoculated mice. The tumor grew quickly and the volume of the tumor reached 12000 mm~3 within 34 days. All the mice died within 40 days and their mean survival time was 37 days. Conclusion A PSCA-expressing tumor model in mice has been successfully established. It can be used to evaluate the activities of drugs or vaccines.

Objective To study the role of protective antigen(PA)and N-terminal segment of lethal factor (LFn)in the entrance of EGFP(enhanced green fluorescent protein)into HeLa cells. Methods The DNA fragments encoding LFn and EGFP were amplified,respectively,and cloned into the plasmid pET-21 a(+)one after another to construct a recombinant plasmid pET-LFn-EGFP. The plasmid was txansformed into BL21 cells to express LFn-EGFP protein under the induction of IPTG. The protein was purified by Ni chelating chromatography. After incubation with LFn-EGFP in the presence of PA or not, the HeLa cells were analyzed by flow cytometry or laser confocal microscopy. Results The fusion protein LFn-EGFP was purified by over 90% homogeneity and retained the ability of LF to bind with PA when incubated with J774A.1 macrophage cells,and could get into HeLa cells. Conclusion The LFn-EGFP could enter the HeLa cells in a PA independent pathway. But PA could help more LFn-EGFP molecules enter into HeLa cells.

Objective To evaluate the activation patterns in the cortexes of expressive aphasics after stroke so as to explore the pathogenic mechanism of expressive aphasia.Methods Blood oxygen level dependent functional magnetic resonance imaging (BOLD-fMRI) was the method of choice.It was administered to 9 subjects with expressive aphasia at 1 to 3 months post-stroke onset and to 10 healthy controls.Active areas in the patients' brains were observed using a block-designed picture-naming task,and language function was tested with the China Rehabilitation Research Center's aphasia examination (CRRCAE).The control group received BOLD-fMRI only.SPM8 software was used to process the fMRI data.Results Differences were observed in the mapping of activated areas between the two groups,but many activated areas showed no difference.Significant differences in activation were observed in areas associated with vision,language and cognition,including the bilateral inferior frontal gyrus,the bilateral superior temporal gyrus,the bilateral insula,the bilateral basal ganglia,the left superior frontal gyrus,the left middle frontal gyrus,the left precentral gyrus,the left thalamus,and the left middle temporal gyrus.All the patients had activated cortex regions associated with visual processing in the left and/or right hemisphere,such as the middle frontal gyrus,the middle temporal gyrus,the lingual gyrus and the fusiform gyrus.The activation volumes in the left hemisphere were significantly smaller than those in normal adults.Regions related to language such as the left inferior frontal gyrus (Broca's area),the left middle frontal gyrus,and the right inferior frontal gyrus (the mirror region of Broca's area) were activated in some of them.While the activation frequency,activated volume and activation intensity generally were all less in the patients than in the controls,the activation intensity in the right superior temporal gyrus,the bilateral superior parietal lobule and the left inferior temporal gyrus were stronger.Conclusions Language production may be associated with multiple,interconnected regions.The right hemisphere participates in natural language processing.Aphasia damages both linguistic and cognitive areas,reducing activation in Broca's aphasia.Activation areas in the left hemisphere and the right inferior frontal gyrus decrease significantly,while some regions in the right hemisphere are relatively more activated.The right inferior frontal gyrus may play a different role in language recovery at different periods of aphasia after stroke.

Objective To evaluate the value of CT virtual endoscopy(CTVE)for microsurgery of pituitary microadenomas via transsphenoidal approach.Methods In 18 patients with pituitary micmadenomas underwent microsurgery via transsphenoidal approach,the presurgical CT data was transfered to work station,the anatomies of the sphenoid sinus were then reconstructed by CTVE.The CTVE images were used to make the preoperative planning and indentify sphenoid sinus, sellar floor,and the position of the tumor during surgery.The CTVE images and the views from intraoperative microscope were compared. Results CTVE could display the anotomoties of sphenoid sinus in a three-dimentional mode.The display rates of superfical antomies such as sphenoid septa,sellar floor,carotid prominence,optic prominence and opticocarotid recess were 344.4%,100%,41.7%,36.1%and 58.3%respectively and the visual fields of these anatomic landmarks were larger than the views from the intraoperative microscope. CTVE could depcit the anatomies of the enhanced carotid ateries and pituitary tissue and some optic canals underlying the sellae by transparent function or heighten the threshold.The sites of the tumor could be marked correctly on CTVE images.CTVE could simulate the operative approach and some operating procedures properatively. Conclusion CTVE can display the anatomies of sphenoid sinus in a three-dimensional mode. In transsphenoidal treatment of pituitary microadenoma,CTVE can help to make preoperative planning,locate the intraoperative structures and make a precise bone window during surgery.

Objective To screen the factors that affect severity of language disorder in subcortex aphasia, and analyze its characteristics and therapeutic effect. Methods 10 moderate degree and 12 severe degree subcortex aphasics were evaluated with Chinese Rehabilitation Research Center Aphasia Examination (CRRCAE) and the Boston Diagnostic Aphasia Examination (BDAE) before and 4 weeks after language training. The factors affecting severity of subcortex aphasia and characteristics were analyzed before training, and effect of language training was examined before and after training. Results Severity of subcortex aphasia was related to age and the complication of apraxia of speech. There was a relatively high level in listening comprehension, reading, repeating and reading aloud abilities. After training, all kinds of language abilities improved significantly in moderate degree subcortex aphasics. Whereas, comprehension and verbal communication capabilities increased obviously, but written communication abilities advanced rarely in severe degree subcortex aphasics. Conclusion Age and the complication of aphasia of speech may be important factors influencing severity of language disorder in subcortex aphasia. There are specific clinical features in subcortex aphasia. Almost all language abilities can be improved by language training, but therapeutic effect is differrent between moderate degree and severe degree subcortex aphasics.

Objective To evaluate the application of multi-parameter three-dimension arterial spin labeling (3D-ASL) in observing the brain perfusion of patients with transient ischemic attack (TIA). Methods A total of 42 TIA patients, admitted to our hospital from July 2014 to March 2017, were included in this study. All subjects underwent conventional MRI, diffusion-weighted imaging (DWI), magnetic resonance angiography (MRA) and 3D-ASL scanning. Abnormal signals, and cerebral arterial stenosis or occlusion were observed under MRI, DWI and MRA; cerebral blood flow (CBF) map was drew after analyzing the 3D-ASL imaging, and abnormal reperfusion of ASL-CBF was qualitatively and quantitatively analyzed. The detection rate of abnormal reperfusion in TIA patients by 3D-ASL (PLD=1.5 s, PLD=2.5 s) and MRA were compared. Results Forty-two TIA patients showed no positive findings on conventional MRI and DWI maps, of which 18 patients showed different degrees of cerebral artery stenosis on MRA maps. Twenty-seven patients (PLD=1.5 s, 64.29％) and 21 (PLD=2.5 s, 50％) on ASL-CBF maps showed different sizes and degrees of abnormal hypoperfusion, and significant difference was found in detection rate of hypoperfusion by 3D-ASL (PLD=1.5 s and PLD=2.5 s, χ2=23.333, P=0.000). The detection rates of hypoperfusion by 3D-ASL (PLD=1.5 s and PLD=2.5 s) were 中华神经医学杂志2017年12月第16卷 第12期 Chin J Neuromed, December 2017, Vol.16, No.12 significantly higher than that by MRA (χ2=17.500, P=0.000; χ2=31.500, P=0.000). Conclusions The 3D-ASL can quantitatively analyze the degrees of perfusion of patients with TIA. 3D-ASL can comprehensively reflect the perfusion status in patients with TIA, and short PLD 3D-ASL is more sensitive than long PLD ASL in finding TIA, while long PLD 3D-ASL can reflect the perfusion status more truly.

Ohjective To explore the impact of constraint-induced aphasia therapy (CIAT) on language function and neural activity in patients with chronic Broca's aphasia.Methods Two chronic aphasics whose use of language was recovering after standard language therapy were selected to receive 1.5 h of CIAT twice daily for two weeks (30 hours in total).Before and after the CIAT they were tested using a block-designed picture-naming task,fMRIs were taken and their use of language was examined.Results The language function assessments showed relatively large improvements in the subjects' use of language after 4 weeks of conventional language training,but not much further change after 8 and 12 weeks.After the two weeks of CIAT,their language function improved further to a certain extent.fMRI showed increased activation in the left inferior frontal gyrus with or without enhanced activation in the left hemisphere,and reduced activation in the right inferior frontal gyrus.Conclusions Functional reorganization induced in the brain by CIAT was associated with up-regulation of the left inferior frontal gyrus or down-regulation of the right inferior frontal gyrus.Even the whole language network may have been modified.