Aim to explore an assay for the early diagnosis of Toxoplasma infection. Methods Free streptavidin was used to attach a biotinylated DNA to the biotinylated second antibody, through amplication of the DNA label, antigen which bound with antibodies was detected, thus Toxoplasma circulating antigens Immuno-PCR assay was estabished. detecting the CAg in serial dilutions of serum and the dynamic variation of serum CAg of Toxoplasma in experimental infected mice using Immuno-PCR and ELISA parallely to compare the sensitivity Result The detection limit of the immuno-PCR was at about 1/1000 dilution, it was at 1/5 dilution for ELISA. CAg of Toxoplasma could be detected as early as the third day after infection using Immuno - pcr, Wheareas the ELISA on the same samples, revealed no detectable CAg elevations until fifth day after infection. Conclusion In comparison with CAgdetecting by ELISA parallelly performed, this method is more sensitive than ELISA, The detection limit of the immuno-PCR was an 200 fold improvement compared with a conventional ELISA. the positive result could be detected earlier than that of ELISA,It provide scientific basis for early diagnosis of Toxoplasmosis.

Objective To investigate the effect of ganoderic acid A( GA-A) on apoptosis, invasion and KDR expression of human U251 cells.Methods Ganoderic acid A( GA-A) was prepared, human U251 cells were treated with 0.1, and 0.5 mmol/L GA-A, and the experiment was divided into blank control, low concentration and high concentration group.The expressions of KDR mRNA and KDR protein was assayed by RT-PCR and Western blot.The effect of GA-A on the proliferation and invasion capability of U251 cells was determined by CCK-8 and transwell assay in vitro, respectively.Flow cytometry was used to detect the influence of GA-A on the cell cycle and apoptosis of U251 cells, and TUNEL staining was detected the cell apoptosis too.Results Compared with the control group, KDR mRNA and protein expression of high concentration and low concentration group were significantly decreased(P <0.05), GA-A can significantly reduce the cell growth rate, reduce the proportion of cells in G1 phase and increase the proportion of S phase and G2 /M phase,cells apoptosis was significantly increased in the high concentration and low concentration group ( P <0.01), and cells proliferation and invasion was significantly decreased (P <0.05).Compared with low concentration group, the high concentration group induce cell apoptosis and inhibit the expression of KDR more significant (P <0.05). Conclusions Ganoderma acid A can induce apoptosis in U251 cells, inhibit proliferation and invasion, and can inhibit the expression of KDR mRNA and protein, which may be one of the mechanisms of anti-tumor.

BACKGROUND:It has been demonstrated that there are different expressions of fatty acid binding proteins (FABPs) in most malignant tumors such as breast cancer, prostate cancer, liver cancer, lung carcinoma and bladder carcinoma;therefore, FABPs are closely related to the occurrence, metastasis, invasion and drug resistance of malignant tumors. OBJECTIVE:To investigate the effects of fatty acid binding protein-5 (FABP-5) silencing on the proliferation, invasion and apoptosis of U251 cel s (human glioma cel s). METHODS:siRNA molecules targeting the mRNA of FABP-5 was designed and chenical y synthesized, which was transiently transfected into U251 cel s. U251 cel s were divided into three groups:Lv-shRNA-FABP-5 was added into FABP-5-shRNA group, LV-shRNA-NC added into negative control group, and blank control group underwent normal culture. mRNA and protein expressions of FABP-5 were detected by RT-PCR and western blot assay, respectively. The cel proliferation in vitro was determined by cel counting kit-8 assay, the cel cycle, apoptosis and expressions of CD44+and CD133+were detected by flow cytometry, and the apoptosis was observed using TUNEL staining. RESULTS AND COUNCLUSION:mRNA and protein expressions of FABP-5 in the FABP-5-shRNA group were significantly lower than those in the negative control and blank control groups. Compared with the negative control and blank control groups, the cel growth rate was significantly decreased, the cel cycle arrested in the G0/G1 phase, and the cel number in the S phase was decreased, and moreover, the proportion of CD44+/CD133+expression was significantly decreased in the FABP-5-shRNA group (P<0.05). Besides, compared with the negative control and blank control groups, the apoptosis rate was significantly increased, and the cel proliferation and invasiveness were significantly decreased in the FABP-5-shRNA group (P<0.05). In conclusion, it is possible that FABP-5 directly or indirectly regulates the cel cycle and apoptosis of glioma cel s, and its expression changes share a close relationship with the invasiveness of tumor cel s.

AIM: To decide the effect that selected siRNA degrades mRNA of IL-1? specifically and suppression of its expression after connected with target site with homology complementary sequence. METHODS: Synthesized DNA expression box aimed directly at target site through PCR reaction in vivo was purified, and transfected into lymphocytes stimulated by LPS. siRNA was transcribed by cellular endogenous RNA polymerase Ⅲ and then evoke the degradation of target mRNA. After 48 hours of transfection, the cell culture supernatant was collected and the concentration of IL-1? was assayed using ELISA. RESULTS: Compared with blank-control and negative-control, selected sequence decreased the expression of IL-1?. Rate of the suppression was about 15%. CONCLUSION: RNAi technology produces specific interference effect in mouse spleen lymphocytes in original culture and inhibits the excretion of IL-1?.

Objective To study the clinical effect of microsurgical surgery for the treatment of intracranial aneurysms in the acute phase. Methods 88 patients with intracranial aneurysm who were treated with the microscopic surgery in our hospital were selected as the research object. The prognosis was evaluated by Glasgow scale ( GOS) ,and the mortality rate during the follow-up period and postoperative complications were observed. Results All the patients received surgery success. The operation time was (60. 5 ± 20. 3) min,and the intraop-erative blood loss was (45. 2 ± 21. 5) mL. 12 months after operation,according to the GOS score of daily living ability,42 patients were of good prognosis and the other 46 cases were of poor prognosis. The good prognosis rate was 47. 7% and it is significantly higher than that of 3 months and 6 months after surgery. The difference was statistically significant (P<0. 05). There were 4 cases died among the 88 patients with a mortality of 4. 5%. Conclusion Using microsurgical treatment to treat intracranial aneurysms can receive good prognosis and low mortality rate. Complications were significantly improved after symptomatic treatment.

Objective To study whether abnormalities can be detected by MR diffusion tensor imaging (DTI) technology in radiologically isolated syndrome (RIS) patients with normal-appearing white matter (NAWM).Methods Twenty-seven patients who met diagnostic criteria for RIS were collected.Sixteen age-and sex-matched healthy controls with normal neurologic examination findings and no history of neurologic or psychopathic disorders were included.All subjects were examined by both conventional scan and DTI scan on GE Signa 3.0 T MRI.All the images were transmitted to the Advantage Workstation 4.2P and postprocessed using functool software.Anisotropic maps and average diffusion coefficient (ADC) maps were reconstructed.Two ROIs were selected in the genu and splenium of the corpus callosum separately at the trigone of lateral ventricle level.Other 2 ROIs were selected in the anterior and posterior body of the corpus callosum separately at the level of the lateral ventricle body,and 1 ROI was selected in the white matter of bilateral frontal and occipital lobe separately,then the FA and MD values were measured.The diffusion indices (FA and MD)were analysed by SPSS 13.0.Independent-sample t test was performed to examine the group differences in each subregion.Intraclass correlation coefficient analysis was performed to assess the diffusion indices of two measurements in each subregion.Results The ICC of 2 measurements was 0.934-0.989 (P ＜ 0.01),which showed favorable consistency.The FA values were decreased obviously in the genu,anterior and posterior body of the corpus callosum of the RIS patients compared with controls (0.705 ±0.040 vs 0.738 ±0.045,0.632 ±0.043 vs 0.675 ±0.042,0.628 ±0.043 vs 0.666 ± 0.045,t =-3.526,-4.487,-3.890,P ＜0.01),but the FA values of the two groups did not show any significant difference in the splenium,the white matter of the frontal and occipital lobe respectively (t =-1.387,-0.683,-1.243,P ＞0.05).In comparison with controls,the RIS patients had increased MD values in the genu,anterior and posterior body of the corpus callosum.(0.891 ±0.038 vs 0.874 ±0.035,0.839 ± 0.047 vs 0.794 ± 0.031,0.833 ± 0.039 vs 0.792 ± 0.057,t =2.101,5.836,5.146,P ＜ 0.05),but the MD values of the two groups did not show any significant difference in the splenium,the white matter of the frontal and occipital lobe respectively (t =1.671,1.702,1.624,P ＞ 0.05).Conclusion The NAWM abnormalities in the patients with radiologically isolated syndrome could be detected by DTI.

Objective To compare the clinical features between radiological isolated syndrome (RIS) and classical multiple sclerosis (CMS), in order to improve the understanding of the RIS. Methods All 35 patients with RIS and 32 patients with CMS were selected. The epidemiological and clinical findings, cerebrospinal fluid, neural electrophysiological examination and magnetic resonance imaging (MRI) data were analyzed. Results There were no statistical differences in sex ratio and onset age between RIS patients and CMS patients (P>0.05). The main symptoms of in patients with RIS were headache (45.7%, 16/35), dizziness (40.0%, 14/35), hypomnesis (20.0%, 7/35) and psychiatric disorders (11.4%, 4/35). But the main symptoms of in patients with CMS were limb weakness (75.0%, 24/32), sensory abnormalities (68.8%, 22/32) and ocular symptoms (34.4%,11/32). The incidences of limb weakness, sensory abnormalities and ocular symptoms in patients with CMS were significantly higher than those in patients with RIS:75.0%(24/32) vs. 0, 68.8%(22/32) vs. 0 and 34.4%(11/32) vs. 0, and there were statistical differences (P<0.01). The 18 patients with RIS and 21 patients with CMS underwent the examination of cerebrospinal fluid, and there was no significant difference in leukocyte between patients with RIS and patients with CMS (P>0.05). The cerebrospinal fluid protein and the incidences of IgG index>0.7 in patients with RIS were significantly lower than those in patients with CMS:0.175 (0.03-0.69) g/L vs. 0.440 (0.04-1.09) g/L and 3/18 vs. 47.6%(10/21), and there were statistical differences (P<0.05). The 15 patients with RIS and 22 patients with CMS underwent the examination of neural electrophysiological, and the abnormality rates of visual evoked potential (VEP) and brain stem auditory evoked potential (BAEP) in patients with RIS were significantly lower than those in patients with CMS:4/15 vs. 63.6%(14/22) and 3/15 vs. 54.5%(12/22), and there were statistical differences (P<0.05). But there was no statistical difference in the abnormality rate of somatosensory evoked potential (SEP) between patients with RIS and patients with CMS (P>0.05). On MRI, the demyelinating lesions of RIS and CMS were both mainly distributed in the periventricular, semi-oval center, infratentorial white matter, partly involving corpus callosum or cortical. The rates of demyelinating lesions in brainstem and cerebellum in patients with RIS were significantly lower than those in patients with CMS:5.7%(2/35) vs. 34.4% (11/32) and 2.9% (1/35) vs. 25.0% (8/32), and there were statistical differences (P<0.01 or <0.05). Comparison with CMS lesions, RIS lesions mainly showed patching and stippled, and there were statistical differences (P<0.01 or <0.05). The rates of lesions enhancement and spinal cord injury in patients with RIS were significantly lower than those in patients with CMS: 2/17 vs. 45.0% (9/20) and 1/14 vs. 43.5% (10/23), and there were statistical differences (P<0.05). Conclusions There are differences in clinical findings, cerebrospinal fluid, neural electrophysiological examination and MRI appearances between RIS and CMS.

Objective To summarize our clinical experience and results of Stanford type B dissection,and analyzed the risk factors of reoperation.Methods From February 2009 to December 2011,81 patients (54 male and 27 female) of Stanford type B aortic dissection underwent surgical procedure in Beijing Anzhen Hospital The mean age was 19 -77 years,averaged (41.6±11.7)years.Associated with hypeftension in 48 cases,Marfan syndrome in 15 case,aortic root aneurysm in 7cases,dilation of aortic sinus and ascending aorta each in one,coarctation of the aorta in 1 case.Reoperation was done in 22cases and 4 of 22 needed the third operation.The risk factors of reoperation were analyzed with Logistic regression analysis.Results Sun (s) procedure was operated on 16 cases,concomitant procedures included Bentall in 7 cases,David in 1,replacement of the ascending aorta in 2 cases,CABG in 1 case.The thoracoabdominal aortic replacement in 31 cases,the descending thoracic aortic replacement in 9 cases,the stented elephant trunk procedure in 24 cases.Concomitant procedures included the bypass from the left subclavian artery to the left common carotid artery in 5 cases,aortic valve replacement and ascending aortic plasty in 3 cases,left subclavian artery reconstruction in 2 cases,double valve replacement in 1 cases,and the bypass from ascending aorta to descending aorta each in one,repair of internal leakage in 1 case.2 patients underwent thoracoabdominal aorta replacement died after surgery due to multiple organ failure caused by postoperative bleeding.The hospital mortality was 2.5％(2/81 cases).Complications occurred in 6 patients (6/81 cases,7.4.％ ),including diastinal bleeding requiring reoperation in 3 cases,respiratory insufficiency and hoarseness each in one,postoperative esophageal fistula needed reoperation and jejunal fistulization in I case.No paraplegia or stroke occurred postoperatively.Logistic regression analysis shows Marfan syndrome is the risk factor of reoperation.Conclusion The surgery on Stanford B aortic dissection can achieve satisfactory clinical results and Marfan syndrome is the risk factor of reoperation.The mid- and long-term results need the further follow-up.

A 43-year-old female presented with multiple, painful, erythematous papules and nodules on the dorsal of the body over the past 17 years. The tumor was painful after cold stimulus and the significant symptoms improved for recent 3 years. Histopathological examinations confirmed the clinical diagnosis of cutaneous leiomyomas. A surgical procedure was one of the therapeutic options in the case of multiple and painful leiomyomas.

Objective Optimal detection of deep hypothermic circulatory arrest (DHCA)-induced early brain injury is important but the effective technique is still not available in the present.The relationship between diffusion wcighted imaging (DW1) and histopathological changes in DHCA-induced piglet brain injury model were analyzed.Methods Eighteen pigs underwent deep hypothermic circulatory arrest and were divided into three groups:group A (n =6) served as control,only underwent anesthesia and thoracotomy,without extracorporeal circulation.Group B (n =6) served as underwent deep hypothermic circulatory arrest at 20 ℃ for 120 min,survived for 1 day.Group C (n =6) served as underwent deep hypothermic circulatory arrest at 20 ℃ for 120 min,survived for 2 days.Diffusion-weighted imaging and histopathology were used to study the brain injury.Results There were hematoxylin-eosin (+) and terminal deoxynucleotidyl transferase biotin-dUTP nick end labeling (+) cells in cerebral cortex and hippocampus.Hyperintensity was most evident in the cerebral cortex in group B and group C with diffusion-weighted imaging,but it was not seen in hippocampus in both groups.There were 5 and 6 piglets can be seen hyperintensity with diffusion-weighted imaging sequence in group B and group C respectively.Hyperintensity was also seen in T2WI sequence in 3 and 5 piglets in group B and group C respectively.Conclusion The study demonstrates the feasibility of diffusion-weighted imaging on evaluation of brain injury after deep hypothermic circulatory arrest and cerebral cortex was selectively vulnerable to cell injury.It is superior to conventional imaging.