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.

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 expression of Pin1 in human pancreatic carcinoma as well as adjacent tissues and to discuss the role of Pinl in oncogenesis of pancreatic carcinoma. Methods Specimen of pancreatic carcinoma tissues and adjacent tissues were collected from 20 cases. Pin1 mRNA and protein expression in pancreatic neoplasm and corresponding adjacent nontumorous tissues were detected by real-time quantitative reverse transcription-polymerase chain reaction (RQ-RT-PCR) and western blot. Results Pin1 was overexpressed at mRNA and protein level in pancreatic carcinoma tissues compared with that in their nontumorous counterparts ( 2.78 ± 1.02 vs 4.36 ± 1.27;5. 48 ± 1.69 vs 9.97 ± 1.86, P ＜ 0.05 ). Pin1 expression was not correlated to clinical stage and pathological grading of the carcinoma. Conclusion Pin1 overexpression may play a key role in pancreatic carcinoma.

Objective Polyinosinic-polycytidylic acid (poly IC) plays an important role in the central nervous system damage and repair.This study was to investigate the effect of poly IC on inflammatory response after spinal cord ischemia-reperfusion injury (SCIRI) in rats.Methods A total of 72 healthy adult male SD rats were equally randomized into a sham-operation, an ischemia-reperfusion (IR), and a poly IC group.The abdominal cavities of the rats were cut open and closed again in the sham-operation group and SCIRI models were established in the IR and poly IC groups by clamping the abdominal aorta, followed by reperfusion 60 minutes later and intraperitoneal injection of saline (0.1 mL) and poly IC (1.25 μg/g), respectively.At 6, 24, and 48 hours after modeling, BBB scores were obtained and the contents of TNFα, IL-1β and IFN-β were measured by ELISA.At 48 hours, the expressions of NF-κB and IL-10 were determined by immunohistochemistry, the area of ischemic necrosis in the spinal cord tissue was calculated by TTC staining, and its morphological changes were observed under the optical microscope.Results At 48 hours after modeling, the BBB scores were significantly lower in the IR and poly IC groups than in the sham-operation group (3.80±0.75 and 9.40±0.49 vs 20.00±0.00, P<0.01), though higher in the poly IC than in the IR group (P<0.01).The rats of the IR group showed extensive degenerated neurons in the gray substance of the spinal cord, with scattered foci of bleeding and blood coagulation, while those of the poly IC group exhibited fewer necrotic neurons and basically normal nuclear morphology, though with a few swelling cells.The ischemic necrosis area of the spinal cord tissue was significantly reduced.The expression of NF-κB was decreased while that of IL-10 increased markedly.Compared with the IR group, the poly IC group showed a significant increase in the expression of IFNβ (117.23±6.06 vs 55.65±4.02, P<0.01) and a remarkably decrease in the expressions of TNFα (190.45±4.16 vs 201.82±2.18, P<0.01) and IL-1β (39.27±2.48 vs 50.59±1.47, P<0.01) at 48 hours.Conclusion Poly IC can protect the spinal cord and reduce inflammatory response after spinal cord ischemia-reperfusion injury.

ObjectiveTo check the isolated heart by coronary angiography to discover the location, na-ture and degree of the coronary artery lesions more accurately and increase the comprehensive evaluation ability of cardiovascular disease.MethodsTen fresh isolated hearts with different causes of death were extracted and injected with barium sulphate as contrast substance by ring injector, then developed under Xper FD20 angiography equipment. The obtained pictures and image data were handled by three-dimen-sional angiography images with the software attached to the angiography equipment. The coronary artery tissues were HE stained and observed by microscope. The HE staining results were compared with the angiographic results.ResultsThe imaging data obtained from the 10 cases for examination showed 8 cases without coronary artery stenosis and 2 cases with Ⅲ, Ⅳcoronary artery stenosis, which were consistent with HE staining results of coronary artery organization and the both results were confirmed. ConclusionIsolated coronary angiography has an unique advantage for accurate grading of classification of coronary artery stenosis, examination of vascular malformation and tiny lesions, which can provide reference for the localization of small lesions and basis during the autopsy for identification conclusion.

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Objective To introduce Checklist Individual Strength(CIS)into China,and evaluate its reliability and validity.Methods CIS was translated into Chinese language and the reverse translation was done by several experts.Validity and reliability were implemented in 214 cases with cerebral infarction.Results 4 components were extracted in factor analysis,and the total cumulative contribution was 78.984%.By correlation analysis,the twenty items were divided into 4 domains:subjective feeling of fatigue,concentration,motivation and physical activity,it was same as in the English version.Cronbach α of four domains were 0.9256,0.9072,0.7598,and 0.9157,respectively.Conclusion Psychometric properties(reliabilities and validities)of CIS Chinese version is met with satisfaction and seems to be adaptable to Chinese cerebral infarction patients.

Objective To analyze the instruments for post stroke fatigue, Fatigue Severity Scale (FSS)，Checklist Individual Strength (CIS) and Fatigue Impact Scale (FIS). Methods The FSS，CIS and FIS were applied in 214 cases with cerebral infarction. They were combined together at the (sub)scale level, and evaluated with Mokken Scale Analysis for Polytomous Items, for validity and reliability. Results and Conclusion Three scales were and The combined scale proved to be acceptable (H>0.5). 3 components were extracted in factor analysis, and the their total cumulative contribution was 77.504%, with reliability (α) of 0.8097，0.7094 and 0.8019 respectively. FSS put emphasis on the fatigue experiences of stroke patients, CIS on the influence of fatigue on, and FIS on the social function.

Objective To analyze the potential factors affecting the prognosis of patients with sepsis and evaluate their values in predicting the disease outcome. Methods A clinical prospective study was conducted. Fifty-three septic patients admitted to intensive care unit (ICU) of the First Affiliated Hospital of Guangzhou Medical University from October 17th, 2012 to August 8th, 2013 were enrolled, and in the same term 35 volunteers having passedphysical check-up were assigned in the healthy control group. According to the severity of the patients, they were divided into sepsis, severe sepsis and septic shock groups. Furthermore, based on the difference in scores of acute physiology and chronic health evaluation Ⅱ (APACHE Ⅱ), the patients were divided into low-risk (APACHE Ⅱ scores < 10), middle-risk (APACHE Ⅱ scores 10 - 19) and high-risk groups (APACHE Ⅱ scores≥ 20 ). According to whether the coagulation dysfunction occurred or not and whether the dysfunction was adjusted or not at the end of observation, the septic patients were divided into non-coagulation defect group, and adjusted and non-adjusted coagulation defect groups. After entrance of groups, the clinical data, including gender, age, body temperature, blood pressure, prothrombin time (PT), prothrombin time activity (PTA), activated partial thromboplastin time (APTT), international normalized ratio (INR), levels of D-dimer, fibrinogen (Fib), blood lactate, and serum procalcitonin (PCT) were recorded, and APACHEⅡscore was calculated. In 24 hours after admission, blood samples were collected, and the levels of interleukins (IL-6, IL-8) were tested by Bio-Plex 200 System. The prognostic factors related to sepsis were screened by binary multivariate logistic regression analysis. The receiver operating characteristic curve (ROC) were used to evaluate the prognostic values of blood lactate, PCT, IL-6, IL-8 and APACHE Ⅱ scores for patients with sepsis. Results Fifty-three patients were enrolled, including 17 patients in sepsis group in which blood coagulation dysfunction occurred in 8 cases, of them 7 being adjusted, and 5 died; 15 patients in severe sepsis group, in which blood coagulation dysfunction occurred in 7 cases, of them 2 being adjusted, and 7 died;21 patients in septic shock group in which blood coagulation dysfunction occurred in 18 cases, of them 4 being adjusted, and 18 died. Both IL-6 and IL-8 levels of sepsis group were significantly higher than those in healthy control group (both P<0.01). Univariate logistic regression analysis showed that the grade of sepsis severity, APACHEⅡscore, IL-6, IL-8, blood lactate, PCT and coagulation dysfunction were related to the prognosis of septic patients. Multivariate logistic regression analysis confirmed that blood lactate concentration [β=0.891,χ2 = 5.872, P = 0.015, odds ratio (OR) = 2.438, 95% confidence intervals (95%CI) was 1.186 - 5.013] and coagulation function status (non-adjusted coagulation defect group:β=3.563,χ2=9.980, P=0.002, OR=35.286, 95%CI was 3.868-3.563) were independent prognostic factors of septic patients. The ROC curve analysis showed:for the level of blood lactate, the area value under ROC curve (AUC) was 0.767, the best cutoff value was 2.15 mmol/L with the Youden index of 0.386;for PCT, the AUC was 0.698, the best cutoff value was 9.39μg/L with the Youden index of 0.406;for IL-8, AUC was 0.686, the best cutoff value was 20.06 ng/L with the Youden index of 0.312;and for IL-6, AUC was 0.681, the best cutoff value was 45.44 ng/L with the Youden index of 0.406. Multivariate logistic regression analysis confirmed that the plasma levels of IL-6 and IL-8 were the independent risk factors of septic patients' coagulation function. The IL-6 concentration of non-coagulation defect group was significantly lower than that in adjusted coagulation defect group (ng/L:29.26 vs. 67.98, P<0.05) and non-adjusted coagulation defect group (ng/L:29.26 vs. 128.00 P<0.05). There was no significant difference in IL-6 level between the adjusted and non-adjusted coagulation defect groups (P>0.05). The IL-8 level of non-coagulation defect group was significantly lower than that of adjusted (ng/L:24.67 vs. 27.23, P<0.05) and non-adjusted coagulation defect groups (ng/L:24.67 vs. 60.14, P<0.05). There was no statistically significant difference in IL-8 concentration between adjusted and non-adjusted coagulation defect groups (P>0.05). Conclusions The grade of sepsis severity, APACHEⅡscore, whether existence of coagulation dysfunction being present or not and whether its presence being adjusted or not during the septic patients' stay in ICU, the levels of blood lactate, PCT, IL-6 and IL-8 on the first day in ICU are significantly correlated to the prognosis of septic patients. Whether the existence of coagulation dysfunction being present or not, whether coagulation dysfunction being adjusted or not and the blood lactate level are the independent prognostic factors of septic patients, and the plasma concentrations of IL-6 and IL-8 are the independent affecting factors of whether coagulation dysfunction occurring or not, therefore they have predicting value concerning the occurrence of coagulation dysfunction in septic patients.