Objective To construct the nuclear receptor-related factor 1 ( Nurr1) specific short-hairpin RNA (shRNA) expressing vectors and study their effects on the expression of endogenous genes in MN9D dopaminergic cell lines. Methods Two RNAi recombinant plasmids (named pSC-N1 and pSC-N2) targeting at Nurrl gene were constructed. The purified plasmids were identified by DNA sequencing. After being transfected into MN9D dopaminergic cell lines with Lipofectamine?2000, the silencing effects of Nurrl shRNA expressing vectors on the expression of Nurrl mRNA and protein in MN9D cells were detected using real time fluorescence quantitative PCR and Western blot Results DNA sequencing confirmed that the Nurrl shRNA expressing vectors were constructed successfully. Nurrl mRNA expression in MN9D cells was specifically suppressed after the transfection of pSC-Nl and pSC-N2 by 62.3% and 45.6% respectively. Moreover, the expression of Nurrl protein was significantly suppressed by 57.4% and 72. 0% respectively. The control vectors had no silencing effect Conclusions The vectors pSC-Nl and pSC-N2 have been successfully constructed and can specifically suppress the expression of Nurrl mRNA and protein. Nurrl specific shRNA expressing vector may provide a novel applicable strategy for the study of the function of the genes associated with Parkinson disease and the development of dopaminergic neuron.

Objective To investigate the clinical features of acquired immunodeficiency syndrome (AIDS) with neurological complications. Methods Five with neurological complications out of 31 patients with confirmed AIDS in Shanghai area from 1992 to 2001 were analyzed. Results Of the five cases, there were one complicated with vacuolar myelopathy, one vacuolar myelopathy accompanied with dementia, one cerebral tuberculoma accompanied with dementia, one cerebral infarction accompanied with symptomatic epilepsia and trigeminal neuralgia, one polyneuropathy. Two of the vacuolar myelopathy are those showing first occurring manifestations. Conclusion AIDS patients are found at high risk of neurological complications, of which vacuolar myelopathy and AIDS related dementia are more common. If young patients who suddenly suffered from dementia, especially in patients accompanied with opportunistic infections, AIDS should be considered. In these cases serum HIV antibodies should be checked.

The etiology of ischemic stroke in young adults is extensive, and cardiogenic embolism and cervicocephalic arterial dissection are the most common. In recent years, the proportion of atherosclerotic ischemic stroke has increased because of the increased incidence and inadequate control for the traditional vascular risk factors in young adults. However, even after a detailed and comprehensive examination, there is stil about 1/3 unknown etiology.

Objective To explore the effect of glucose regulated protein (GRP)78 on the neuronal apoptosis after cerebral ischemia reperfusion injury in rats. Methods Middle cerebral artery ischemia reperfusion rat's models were used with the modified filament method. The expression of GRP78 in the ischemic penumbra tissue was detected by RT-PCR, Western blot and immunohistochemistry at different time points. Primary cultured rat's neurons were exposed to hypoxia and subsequently reoxygenation. Western blot was used to investigate the expression of GRP78. The changes of the neuronal apoptosis after overexpression of GRP78 induced by 2-deoxyglucose were detected. Results The expression of GRP78 in the ischemic penumbra tissue in model group was significantly increased (mRNA : 0.7367±0.0651, F= 477.160, P < 0.01 ; Protein : 0. 8129±0. 0748, F=39.857, P < 0.01). The neuronal survival status was increased after overpression of GRP78 (increased by 39.22% ± 0. 44%, t=46.374, P < 0.01) while the neuronal apoptosis was decreased (decreased by 16.60±1.02, t=7.530, P <0.01). Conclusion Focal cerebral ischemia reperfusion can induce endoplasmic reticulum stress which plays a role in the neuronal apoptosis. The increased expression of GRP78 may protect the ischemic tissue from neuronal apoptosis.

Objective To investigate the effect of Akt pathway inhibitor AZD5363 on cell proliferation and invasion of QBC939 and RBE cholangiocarcinoma cells and the mechanism.Methods Western blotting was used to detect Akt and downstream protein and mTOR protein expression in two cancer cell lines after process by AZD5363.Inhibition rate and cytotoxicity was tested by CCK-8 assay,and Transwell assay was used to evaluate the invasive ability of cancer cells.Results QBC9393 cell exposed to AZD5363 LD50 drug concentration (24 ±9) was significantly different compared with control group (t =4.47,P ＜ 0.05),RBE cells LD50 drug concentration (21 ± 8) was significantly different compared with control (t =4.41,P ＜ 0.05).Tumor invasion capacity of QBC939 in drug concentrations of 20 μmol/L (63 ± 12) and 0 μmoL/L (271 ± 27),the difference was statistically significant.RBE exposed AZD5363 upon drug concentrations of 20 μmol/L (58 ± 23) and 0 μmol/L (235 ± 21),the difference was statistically significant.AZD5363 promotes phosphorylation of mTOR in QBC939.Conclusions AZD5363 inhibits the proliferation and migration,inhibiting the phosphorylation of Akt and its downstream molecules.AZD5363 promotes phosphorylation of mTOR in QBC939.

HRP solution (33%) was injected into the left superior colliculus of 21 adult albino rats. 1-2 days after injection, the animal was sacrificed. Brains were frozen sectioned and processed according to. DAB- or TMB-method. The labeled neurones were examined in the right superior colliculus and other areas of the brain. The resu ts are as follows:1. No matter where the HRP was injected (either in the rostral or caudal part of the superior colliculus or thepart between them) HRP labeled neurones were always observed on the opposite superior colliculus if the injection sites reached layers deeper than the stratum oPticum. The locations of the labeled neurones corresponded roughly to the sites of HRP injections. However, no HRP labeled neurones were observed when the core of HRP deposites was restricted to layers superficial to the stratum griseum intermediale.2. Of all labeled neurones, 53.6% were located in the stratum griseum intermediale, 16.5% in the stratum opticum. The rest were in deeper layers. In no case were HRP labeled neurones observed in the stratum zonale or stratum griseum superficiale.3. Labeled neurones could be classified morphologically into vertical fusiform, horizontal fusiform and multipolar neurones.4. In addition to the visual cortex, labeled neurones were also found in the inferior colliculi, paralemniscal nuclei, dorsal nuclei of the lateral lemniscus, substantia nigrae and lateral tegmental nuclei bilaterally. Labeled neurones were also found in the ipsilateral ventral nucleus of the lateral geniculate body, contralatera pretectal area and reticular formation.

The present paper respectively analyzed the results of deep vein ascending phlebography in examining 105 cases(136 extremities)of the lower extrimity venous ulcer. This study showed that communicating branch incompetence eccompanied with deep vein incompetence in 99 lower extremities (72 8%);communicating branch incompetence without deep vein incompetence in 32 lower extremities(23 5%);and saphenous incompetence without communicating branch incompetence in five lower extremities (3 7%). According to the results of phlebography, the pathogenesis of the leg venous ulcer and the selection of operation were discussed in the paper.

In order to investigate the distribution and other features of callosal neurons inthe visual areas,12 adult albino rats were used in a series of experiments,in whichHRP(30～50%,DAB brown reaction)was injected into one hemisphere and theretrograde labelled cells were observed in the other hemisphere.1.Hardly any labelled neuron was found in contralateral visual cortices whenthe injections were made at sites within 3.6 mm from the midline of the brain.However,when the injections were made at sites 4～6 mm from the midline,manylabelled neurons could be observed.2.Generally,most labelled cells aggregated near the border between area 17 and18a.In area 18a numerous cells were labelled,which distributed in the form ofincontinuous patches.In the proper of area 17,however,less labelled cells werefound.3.A gross topographical correlation was observed between locations of injectionand labelling on the two hemispheres.4.The labelled cells were found in all cortical layers except layer Ⅰ.In area 17they were mainly in layers Ⅳ and Ⅴ,while in area 18 a they were found mainly inlayers Ⅲ,Ⅳ and Ⅴ.5.Some callosal neurons were lightly labelled.For these cells,only the outlinesof the cell body could be seen even under dark-field microscope.The diameters ofthese cells were about 8～10?m.A few cells were as large as 16?m in diameter.Other labelled neurons could be seen with both dark-field and bright-field illumina-tions.They were identified as pyramidal,multipolar bipolar and granular cells accordingto the shape of the cell bodies and the pattern of dendrites.The majority of labelledcells were pyramidal cells,with a long apical dendrite to the pia surface,10～20?min diameter.Other ceils were 10～20?m for multipolar cells,and less than 10?m forgranular cells.Comparisons were made between the above results and the physiological studies.

To analyze the main factors influencing the ethical review work of municipal hospitals in Shanxi Province by investigating the current situation of the construction and operation of ethics committees, and to put forward reasonable suggestions for improving the work of ethics committees. Questionnaire surveys and interviews were used to investigate the composition of ethics committees, systems and standard operating procedures, personnel training, review project methods, time limits, etc. of 24 hospitals at municipal levels in Shanxi Province. The establishment and system of ethics committees in tertiary hospitals at prefecture and city level were basically reasonable, and they could actively play the role of ethics committees. At present, most hospitals do not pay enough attention to the work of ethics committees, lack of full-time staff and systematic training, resulting in insufficient ethics review capabilities. The construction of the ethics committee of the second-level hospital is a mere formality, and no substantive work has been carried out. It is recommended to strengthen the supervision, increase the training of ethics committees and researchers, improve the information management of ethics committees, and establish regional ethics committees to further improve the ability and efficiency of ethics review.

Gene editing technology has been a hotspot in the field of biotechnology. CRISPR/Cas systems are efficient gene editing tools because of its specificity, simplicity and flexibility, these features enabled the rapid application of CRISPR/Cas systems in a variety of organisms. Moreover, the combination of transcriptional activator with dead Cas protein can achieve specific regulation of gene expression at the transcription level, which has made important contributions to the development of biotechnology in medical and agriculture. Overexpression of foreign genes is a common method to verify gene function and regulation. However, due to the limitation of vector capacity, it is difficult to achieve overexpression of multiple genes. CRISPR/Cas9 activation system can regulate the expression of multiple genes under the guidance of different guide RNAs to verify gene functions at the regulatory level. This review summarizes the composition of the CRISPR/Cas9 activation system and different activation strategies, and summarizes solutions for excessive activation. It may facilitate the application of CRISPR/Cas9 activation system in genetic improvement of cotton and herbicide resistance research.
Biotechnology ; CRISPR-Cas Systems/genetics* ; Gene Editing ; Phenotype ; RNA, Guide, Kinetoplastida/metabolism*