10.3969/j.issn.2095-4344.2013.24.022

Objective:To investigate microRNA-29b ( miR-29b) expression in cerebral cortex , spinal cord, fore limb muscle, and serum of SOD1-G93A amyotrophic lateral sclerosis ( ALS) mice, and to identify the biomarker and to assess diagnostic values for ALS .Methods:Cerebral cortex , spinal cord , fore limb muscle and serum from 16 SOD1-G93 A ALS mice and 16 wild-type mice were taken and then microRNA extracted , detecting the expression of miR-29 b by real-time quantitative polymerase chain re-action ( RT-qPCR ) .The diagnostic performance of miR-29b for ALS was estimated by the receiver operating characteristic ( ROC ) curve . Results: The results from the validation indicated that the differences in miR-29b between the cerebral cortex of SOD1-G93A ALS and the healthy control subjects were statistically significant (P=0.001).Meanwhile, the expressions 8, 12, and 16 weeks later were higher than those of the controls ( ALS vs.Control: 8 weeks, P=0.044; 12 weeks, P=0.018; 16 weeks, P=0.045).When the relative expression level of miR-29b was used to diagnose ALS in SOD1-G93A ALS mice, the area under the ROC (area under the curve, AUC) was 0.885, if the diagnostic threshold was set at 0.185 6, the sensitivity and specificity were 92.9%and 71.4%.Conclusion:MiR-29 b may act as medical monitoring indices of ALS in early time .

Some measures about improving the teaching quality in respiratory department training are described in this report,including identifying the aim and importance of major training,raising students'enthusiasm,enhancing the basic major procedure training,performing various teaching,concentrating on the students'response and altering teaching methods continuously

Objective To explore the isometry of grafts in PCL(posterior cruciate ligament)double-bundle re-construction under femoral tunnel shifting condition.Method Knee specimens from ten fresh frozen cadavers were used.PCL were divided into anterolateral bundles(ALB)and posteromedial bundles(PMB)to the inser-tion footorint.The anterior,postedor,proximal,distal and central points of the two bundles'femoral attachment site were respectivelyanchored to the middle of the PCL's tibial attachment site by the trial wires.Changes in length of the intra-articular part of the wires were recorded while the knee was flexed from 0°to 120°.Result The length changes in every point were compared.All of the maximal length changes of ALB's proximal,pos-todor points and PMB's proximal points were not greater than 2mm.No significant difference between the length changes of ALB's proximal point and posterior(P=0.864>0.05)was found.Conclusions The femo-ral tunnel for the PCL double-bundle reconstruction should be located as follows:ALB should be at the middle point of upper edge of femoral attachment site(proximal point),while PIVIB at the middle point of femoral attachment site(proximal point).

BACKGROUND: Foreign studies have demonstrated that the blue light at 470 nm inhibits melatonin secretion and displays the most obvious biorhythm regulation. To date, light-emitting diode (LED) applied in regulating biorhythm remains poorly explored. OBJECTIVE: To explore whether a certain intensity of LED (blue) light could induce retinal injury in rats. DESIGN, TIME AND SETTING: Randomized, controlled animal experiment was performed at the Animal Laboratory of Peking University Third Hospital between May 2007 and April 2008. MATERIALS: A total of 32 SD rats and 16 BN rats were provided by Animal Department of Peking University Third Hospital. METHODS: A total of 16 SD and 16 BN rats were respectively randomly divided into test and control groups. Test group rats were placed in light boxes which were controlled by blue LED (wavelength 470 nm) at a intensity of 300-350μW/cm2, 4 hours everyday for 3 days. The remaining SD rats were placed in light box which was controlled by blue LED (wavelength 470 nm) at a intensity of 120-150μW/cm2, 4 hours everyday for 3 days. The control rats were not treated. MAIN OUTCOME MEASURES: At the second day after light irradiation, the rats of all groups were sacrificed and both eyeballs were harvested. The frozen sections were subjected to hematoxylin-eosin staining to observe changes of rat retina. RESULTS: A total of 48 rats were included in final analysis. The retina of SD rats became thinning and disorderly arranged following blue LED irradiation at density of 300-350μW/cm2, but the retina of BN rat remained unchanged similar to control group. After blue LED irradiation at density of 120-150μW/cm2, the retina of SD rat remained unchanged similar to control group. CONCLUSION: Blue LED light source irradiation at a intensity of 300-350μW/cm2 is safe to pigment-protected retina, and at a intensity of 120-150μW/cm2 does not injury retina of different races of rats.

BACKGROUND: Leucine-rich repeats and immunoglobulin-like domains 1 (LRIG1) gene showed low expression in glioma cells. LRIG1 gene overexpression significantly enhanced LRIG1 mRNA and protein expression and inhibited its biological behavior. However, very few researchs are reported from the stand-point of inhibition of LRIG1 gene expression. OBJECTIVE: To construct specific RNA interference plasmids for LRIG1, establish stably transfected human glioma GL15 cell line, and observe its effect on expression of target gene LRIG1. METHODS: Designed and synthesized two shRNAs (named LRIG1-shRNA1 and LRIG1-shRNA2) specific for LRIG1 mRNA according to the GenBank, and one scrambled shRNA sequence as negative control, named pGenesil2-negative shRNA. The shRNA was inserted into pGenesil2 vector and sequenced. The recombinant vectors were transformed into E. coli. Picked up the positive clones and extracted the plasmids, which were transfected into GL15 cells by Metafectine. G418 was applied to select the stably transfected cell clones. Western Blotting was performed to examine the LRIG1 protein level.RESULTS AND CONCLUSION: The recombinant plasmids which contain shRNA were analyzed by restriction endonuclease digestion and DNA sequence, and it was proved that the fragment was inserted into the expected sites. Compared with the negative control group, the level of LRIG1 protein expression in pGenesil2-LRIG1-shRNA1(LRIG11) transfected cells and in pGenesil2-LRIG1-shRNA2(LRIG12) transfected cells was decreased by 47.9% (P < 0.01) and 32.8% (P > 0.05). The results confirmed that RNAi expressing vector specific for LRIG1 gene (pGenesil2-LRIG1-shRNA1) was successfully constructed, and the stable cell clones transfected with the shRNA expression vector showed inhibition of the expression of LRIG1 in glioma cell line GL15.

This study was focus on investigating the anti-liver fibrosis effects of insulin-like growth factor-1 (IGF-1) in vitro.The effects of IGF-1 on human liver L-02 cell viability and cell cycle were observed.CC14-induced L-02 cell injury was set up to detect the anti-apoptotic activity of insulin-like growth factor-1 (IGF-1).Transforming growth factor β1 (TGF-β1) induced hepatic stellate cell line (HSC-T6) were used as a liver fibrosis model in vitro to analyze the effects of IGF-1 on the expression of liver fibrosis proteins and intracellular TGF-β1/Smad signaling pathway in HSC-T6 cells.The results showed that IGF-1 could relieve the growth inhibition effects of TGF-β1 on L-02 cells,increase the viability of L-02 cells injured by CCl4,decrease the expression of liver fibrosis proteins,and inhibit the TGF-β1/Smad signaling pathway by inhibiting the phosphorylation of Smad3.Our study suggested that IGF-1 exerted anti-liver fibrosis effects by stimulating L-02 cells proliferation,reducing cell damage and inhibiting ECM accumulation via interfering TGF-β1/Smad signaling pathway.

Objective To reveal the influential factors on scalp electroencephalogram (EEG)recording and provide valuable information for localization of the epileptic focus by analyzing the characteristics of spikes with continuous focal periodic discharges on scalp and cortical EEG. Methods Five patients with refractory epilepsy who had low amplitude spikes with continuous focal periodic discharges on interictal scalp EEG were studied. Intracranial EEG recording was also performed in patients. The amplitudes of spikes and cortical areas of spike-wave foci were measured by DaVinci system. Patterns of continuous periodic activity were determined by autocorrelograms,power spectral density and coherence analysis using Matlab and Spike2 software.T-test was employed to compare the mean amplitudes of spikes on the scalp and cortical EEG.Results The amplitudes of spikes recorded on scalp EEG of the 5 patients were:(22.2±4.8),(30.4±7.1),(20.7±3.2),(58.4±10.1),(23.4±3.9) μV.The amplitudes of spikes recorded on cortical EEG of the 5 patients were:(1253.8 ± 199.3),(806.5 ± 161.4),( 1585.7 ±305.7),(922.5 ± 140.6),(736.8 ±70.9) μV.The amplitudes of spikes on scalp EEG were significantly higher than those on cortical EEG ( t =6.394,P ＜ 0.05 ).The cortical areas of spike-wave foci of the 5 patients were:4.0,6.0,3.5,5.5,6.5 cm2.Power spectral density and autocorrelugrams showed 1-3 Hz oscillations on the cortical of spike-wave foci. Cross-correlation and coherence analysis showed synchronization of electrical activity in two contacts of intracranial electrodes. Conclusion The low amplitude spikes with continuous focal periodic discharges on interictal scalp EEG provide valuable information for localization of the epileptic focus.

Objective To study whether bellidifolin has rehabilitation effect on injured sciatic nerve rats,and whether ciliary neurotrophic factor (CNTF) is involved in this mechanism.Methods 225 male wistar rats were made to be sciatic nerve injured models,with right sciatic nerve being cut and sewed under microscopy,left sciatic being sham.Rats were randomly divided into control group,bellidifolin 25 mg,50 mg,75 mg groups and mecobalamin group,with 45 rats in each group.Rats in control group were just injected sodium chloride,others were intra-peritonealiy injected different doses of bellidifolin and mecobalamin after operation.Results Three weeks after operation,SFl of control group,mecobalaming group,bellidifolin 25 mg,50 mg and 75 mg group were-84.35± 4.87,-45.20±2.30,-70.42±4.21,-57.73±3.46 and-64.38±4.38 respectively.Compared with control group,others showed significant differences (P＜0.05).There were statistically differences between bellidifolin groups and mecobalaming group(P＜0.05).Within bellidifolin groups,50 mg group showed difference compared with 25 mg and 75 rg groups(P=0.031).TSW results also showed differences among bellidifolin groups,control group and mecobalaming group.There were statistical differences among bellidifolin groups(P＜0.05).Each groups with immunohistochemistry analysis,CNTF expression showed statistically differences among bellidifolin 50 mg group and 25 mg,70 rg groups,Bellidifolin 50 mg group was higher than others(P＜0.05).Conclusion Bellidifolin can promote the recovery of injured sciatic nerve,especially the concentration of 50 mg bellidifolin,and CNTF is involved in the rehablitation process.

Objective To observe whether bone marrow mesenchymal stem cell transplantation can improve vasospastic rats sense and motor function.Methods Rats grouped with randomized number method as Control group,Subarachnoid hemorrhage group.Stem cell culture media group and Stem cell transplantation group.Subarachnoid hemorrhage model were made with tail artery blood twice injection,2 days after 2' nd injection.Bone marrow mesenchymal stem cell were transplanted to lateral cistern.Subarachnoid hemorrhage(SAH) group didn' t transplant stem cell.Stem cell culture media group injected DMEM media as DMEM group.Stem cell transplantation group injected 30μl Bone Marrow mesenchymal stem cell suspension,so called BMSCs group.Neurofunctional score and learning memory expression were detected with morris mazer and Neurofunctional Score Scale in each group.Results After transplantation for 7 d,functional score of Control,SAH,DMEM and Stem cell group were 3.95 ±2.51,7.20 ± 1.03,7.23 ± 1.79 and 5.81 ± 1.11 respectively.Compared with others groups,Stem cell group score was significantly decrease(P=0.017).After transplanting stem cell for 14 d,the mean spanning plate time in Control group,SAH group,DMEM group and Stem cell group were 7.38 ± 1.73,4.52 ± 0.90,5.11 ± 1.93 and 7.32 ± 2.16 respectively,SAH and DMEM group vs other 2 groups,there were clearly statistically differences (P =0.009),while between control group and stem cell group,there were no statistically differences (P =0.14).Conclusion SAH rat transplant stem cell can improve sense,motor and learning expression in certain level.