Objective　To study the characteristics of serious central nervous system(CNS) involvement in childhood systemic lupus erythematosus.Methods　We made a comparison on the level of ANA、dsDNA and positive rate of Sm、C3 between primary and secondary CNS involvement and analysed the clinical manifestations between two groups.Results　The level of ANA、dsDNA and ositive rate of Sm、C3 were not related with SLE encephalopathy;EEG was useful to the diagnosis of SLE.Conclusion　The differiential diagnosis between primary and secondary CNSD in volvement of SLE must be analysed according to clinical manifestations and other laboratory findings.

Background Neovascular diseases such as retinopathy of prematurity often leads to irreversible vision loss.The study on oxidative damage mechanism is becoming more and more important.Whether brain derived neurotrophic factor (BNDF) has protection to retinal ganglion cells(RGCs) has few research reports.Objective The study was to investigate the expression changes of BDNFin mouse retinas of oxygen-induced retinopathy (OIR).Methods Thirty SPF C57BL/6J immature rats were divided into OIR group and normal control group and to fifteen rats for each group.OIR models were established by raising 7-day-old (P7) mice together with maternal mouse in (75±3) ％ oxygen environment for 5 days and then returned to the normal air environment,and the mice of the normal control group were raised in the normal air environment.The P17 mice were sacrificed for retinal histopathological examination by hematoxylin and eosin staining, and the number of vascular endothelial cell nucleus extending the inner limiting membrane was counted.Whole retinal mounts were prepared after fluorescein isothiacyanate-dextran (FITC-dextran) (9 ml/kg) was retrosorbitally injected,and the distribution of retinal vessels was observed in P17 mice.The relative expression levels of BNDF in retinas were detected in P13, P15, P17 mice, and the results were compared between the normal control group and the OIR group.Results Histopathological examination showed that retinal inner limiting membrane was smooth in the normal control group, but a lots of vascular endothelial cell nucleus extending the inner limiting membrane were seen under the optical microscope in the OIR group.The number of the vascular endothelial cell nucleus extending the inner limiting membrane was 1.70±0.68 in the normal group and 45.3±3.13 in the OIR group, showing significant difference between them (t =86.5, P =0.00).Whole retinal mount revealed that normal retinal vessels and network-like capillaries were exhibited in the mice in the normal group, while tortuous vessels,capillary loss and non-perfusion areas were revealed in the OIR group on the whole retina mounts.The relative expressing levels of BDNF in retinas were 263.992±9.451 and 218.432±9.710 in P15 and P17 mice in the OIR group,which were significantly higher than 230.324±7.779 and 115.846±7.305 in the normal control group (t=14.2,42.3 ,P＜0.05).Conclusions OIR can be inhibited by increasing the expression of BNDF.

Objective:To observe the effects of remifentanil on hemodynamics and plasma angiotentratin-Ⅱ (AgⅡ) level in children with cleft palate undergoing repair operation.Methods:Fifty patients,ASA Ⅰ-Ⅱ, were randomly divided into two groups,remifentanil group (R group) and fentanyl reoup (F group).Patients in R group received remifentanil combined with propofol anesthesia,those in F group received fentanyl combined with propofol anesthesia.Vecuronium bromide was given as muscle relaxant in both groups.ECG,HR,BP and SpO_2 were monitored in all the patients during operation,and peripheral venous blood samples were taken 1 d before operation(T1 ),at the time of endotracheal intubation(T2),10 min after operation beginning(T3),5 minutes after extubation(T4) for the check of plasma AgⅡ level.Results:At T2 and T3 HR,SBP,DBP and plasma AgⅡ level were significant lower in both group of the patients (P

BACKGROUND: Vascular endothelial growth factor receptor 2 (VEGFR2) is primarily involved in vascular endothelial growth factor (VEGF)-mediated signal transduction and plays a critical role in the pathological angiogenesis that occurs in a number of diseases, including leukemia. Besides, VEGF secreted by leukemia cells also induces its own expression which leads to an enhanced production of VEGFR2 which contributes to the survival and proliferation of leukemia cells.OBJECTrVE: To evaluate the inhibitive effect of Lenti6/shVEGFR2 on the VEGFR2 expression and leukemia growth in mouse.DESIGN: A randomized, parallelized, controlled and open trial.SETTING: Department of Pediatrics, the Second Affiliated Hospital of Sun Yat-sen University; Biotechnology Research Center, Sun Yat-sen University.MATERIALS: The experiment had been done in the laboratories for Medical Research Center of the Second Affiliated Hospital, Sun Yat-sen University and Biotechnology Research Center, Sun Yat-sen University from May 2004 to January Lentiviral RNAi Expression System was purchased from Invitrogen, Co.,Ltd.; human VEGFR2 Mcb (PE) was purchased from R&D; CD31 immunohistochemistry kit was purchased from Boster, Co.,Ltd.; CD33-PE fluorescence labeled antibody was purchased from BD, Co.,Ltd.transiently and expression clone (Lenti6/shVEGFR2) was constructed, then cotransfected with ViraPowerTM Packaging Mix pU6/shVEGFR2 entry clone and transducting with Lenti6/shVEGFR2 expression clone, the effect on the development of intravenous xenograft leukemia mouse model, the distribution of microvessels in mouse bone marrow was observed after leukemia model mouse injected with recombinant lentivirus (group B); leukemia model mouse injected with recombinant lentivirus and endothelial cell (group C); leukemia model mouse injected with endothelial cell (group D). Through detecting changes of CD33 positive cells and microvessel density (MVD) in bone marrow, observing peripheral blood cell (PBC)smear and slice of liver, spleen, the effect of Lenti6/shVEGFR2 recombinant lentivirus on mouse leukemia was evaluated.mediated with lentivirus on VEGFNEGFR2 paracrine and autocrine loops in leukemia mouse.effective in inhibiting HL60 cell. pU6/shVEGFR2 entry clone constructed according to it had cell inhibitory rate as high as after transfection of pU6/shVEGFR2 entry clone and transduction of Lenti6/shVEGFR2 expression clone: 48 hours after transfection of pU6/shVEGFR2 entry clone and transduction of Lenti6/shVEGFR2 expression clone, the cell growth inhibitive rates were similar. However, the cell growth inhibitive rate of entry clone descended rapidly after 48 hours (P＜0.01); which of expression clone changed slowly, reaching the peak at 96 hours, dropped slightly, having no significance mouse: The amount of HL60 cells in bone marrow of groups A, B and C detected with flow cytometry were (25.8%±4.9)%, (14.3%±5.1)%, (8.4±2.6)%, respectively (P＜0.05); MVD in group C was obviously less than that in group D (P＜0.05); The amount of HL60 cells in leukemia model mouse injected with recombinant lentivirus and endothelial cell was the lowest as compared with the other groups.

Objective To investigate the effects of macrophage migration inhibitory factor (MIF) overexpression on the expression of interleukin-8 (IL-8),martix metalloproteinase-9 (MMP-9) and invasion of human cervical cancer SiHa cells.Methods Chemical synthesis MIF eDNA gene,designed primer sequence including XhoI and BamHI enzyme sites,MIF gene was amplified by polymerase chain reaction (PCR),constructed eukaryotic expression vector pEGFP-N1/MIF and transfected into SiHa cells using Lipofectamine and won over-expression of MIF.The expression of MIF in supernatant fluid was detected by ELISA,the expression of MIF,IL-8,MMP-9 in both mRNA and protein levels were detected by real-time fluorescence quantitative-PCR and immunocytochemistry respectively.The effect of over-expressed MIF on migration was detected by Boyden small chamber.Results The expression of protein in supernatant fluid transfected with pEGFP-N1/MIF was significantly increased (Fgroup =8267.564,P ＜ 0.01),the expression of MIF,IL-8,MMP-9 in both mRNA and protein in SiHa cells transfected with pEGFP-N1/MIF were significantly increased (F values were 7019.619,2148.094,3303.540,1565.114,2807.300,523.466,P ＜ 0.01),and there was a positive correlation among MIF,IL-8,MMP-9 expression in both mRNA and protein (r values were 0.865,0.895,0.934,0.908,P ＜ 0.01).Invasion ability in SiHa cells transfected with pEGFP-N1/MIF was obviously increased (F=3430.898,P＜ 0.01).Conclusion The over-expression MIF gene in SiHa cells can promote cervical cancer cell invasion and metastasis of ability,which could be associated with the upregulation of IL-8 and MMP-9 expression.

Objective:To establish an HPLC method for the determination of optical isomers in carfilzomib .Methods:The sample was separated on a Chiralpak OX-H column.The mobile phase consisted of n-hexane∶isopropanol∶alcohol (89 ∶5 ∶6, v/v/v) with a flow rat of 1.0 ml· min-1 .The detection wavelength was 220 nm.Results:The linear rang of enantiomer , diastereomer Ⅰand di-astereomer Ⅱwas 0.54-2.14 μg· ml -1,0.11-1.80 μg· ml-1 and 0.11-1.81 μg· ml-1(r≥0.998), respectively.The lower limit of quantification was 0.07-0.27 μg· ml-1 .The recoveries of optical isomers were within the range of 99.6%-100.9% with RSD of 1.13%-1.59%(n=9).Conclusion:The method is sensitive, simple, fast, accurate and specific, and suitable for the study of opti-cal isomers in carfilzomib .

Objective To investigate the role of microRNA-490-5p (miR-490-5p) in the regulation of visceral sensitivity in rats with intestinal dysfunction.Methods The lentivirus vector system was used to construct the rno-miRNA-490-5p lentivirus expression vector.The rats were divided into normal group,diarrhea-predominant irritable bowel syndrome (IBS-D) group,lentivirus empty vector group and the siRNA silent group and the latter three groups were model groups.The efficiency of siRNA was measured by real-time polymerase chain reaction (PCR).The rats were gavaged with 10％ India ink,and then the time of first black stool,water content of feces and threshold of expansion capacity caused abdominal elevation or back arching were calculated.The visceral sensitivity of rats after miRNA-490-5p silenced was evaluated with abdominal withdrawal reflex (AWR) score by stimulating with different intensities of colonic dilatation.The abdominal electrical activity of rats stimulated by colonic distension was measured by BL-420F biological and functional experimental system.The change of the tension of rats isolated colon intestinal stimulated with acetylcholine chloride was also detected by BL-420F biological and functional experimental system.T test was used to compare the differences between the model groups and the normal group.One way analysis of variance was performed for multi-group comparison after miRNA-490-5p interfered.For comparison between two groups among multiple groups,least significant difference (LSD) method was used when the variance was equal,and Games-Howell method was used when the variance was unequal.Results The gastrointestinal propulsion time and the threshold of expansion capacity caused abdominal elevation or back arching of model groups were both lower than those of the normal group ((8.54±4.07) hvs (12.33±2.23) h,(0.56±0.08) mL vs (0.84±0.09) mL),and the differences were statistically significant (t =2.62 and 6.37,both P ＜ 0.05).After distension with 0.8 mL and 1.2 mL sodium chloride solution,the AWR scores of model groups were significantly higher than those of the normal group (3.20±0.56 vs 1.20±0.45,3.73±0.46 vs 2.60±0.55),and the differences were statistically significant (t=7.20 and 4.58,both P＜0.01).There was no significant difference in AWR score between the model groups and the normal group when distended with 1.6 mL sodium chloride solution (3.93 ±0.26 vs 3.80 ± 0.45) (P＞0.05).After miRNA-490-5p silenced,gastrointestinal propulsion time of normal group,IBS-D group,lentivirus empty vector group and the siRNA silent group was (11.12±1.01) h,(6.23±3.17) h,(6.09 ± 2.26) h and (12.36±1.97) h,and the differences among four groups were statistically significant (F=10.55,P＜0.01).The abdominal electrical activity of normal group,IBS-D group,lentivirus empty vector group and the siRNA silent group distension stimulated with 0.8 mL and 1.2 mL sodium chloride solution was (64.91 ± 10.50),(101.79 ±11.73),(80.49±1.27),(66.92±3.24) μV,and (105.09±52.40),(131.71± 16.74),(111.00±6.41) and (95.49± 4.2) μV,and the differences among four groups were statistically significant (F=16.82 and 9.14,both P＜0.05).There was no significant difference in abdominal electrical activity amplitude between silenced group and normal group ((66.92±3.24) μV vs (64.91±10.49) μV and (95.49±4.22) μV vs (105.09±2.40) μV) (all P＞ 0.05).After distension with 1.6 mL sodium chloride solution,the abdominal electrical activity amplitudeof silenced group was lower than the other groups,and the differences were statistically significant (F=11.09,P＜0.01).After adding 1∶1 000 acetylcholine chloride added,the tension of colon of normal group,IBS-D group,lentivirus empty vector group and the siRNA silent group increased by 0.71 ± 0.21,0.81±0.06,0.88±0.21 and 0.43±0.07,however there was no significant difference among the four groups (F=2.57,P =0.100).Conclusions Visceral hypersensitivity existed in rats with intestinal dysfunction.miRNA-490-5p may be involved in the regulation of visceral sensitivity.

BACKGROUND:Hydroxyapatite/gel nano-composite has the same mechanical strength to the natural bone, but its ability to repair bone defects and osteogenic effect need to be confirmed by further studies.
OBJECTIVE:To explore the repair effect of hydroxyapatite/gel bionic composite in skul defects of rabbits.
METHODS: The hole-like calvarium defect models were established in rabbits, and treated with hydroxyapatite/ gel composites (hydroxyapatite/gel group), autologous skul as positive control (autologous bone group) and
nothing as negative control (blank group). The repairing condition in the skul defect areas were observed and analyzed by X-ray and CT at 4, 8, 12 weeks after implantation.
RESULTS AND CONCLUSION: After 8 weeks, X-ray assessment showed that normal-like bone tissue appeared in the defect region of the autologous bone group; in the hydroxyapatite/gel group, dense bone with similar
morphology to normal bone tissue was found in the central site of defect region, and the boundary was slightly blurred. After 12 weeks, the hydroxyapatite/gel showed blurred edge compared with autologous bone, and the center of the composite was disconnected; in the blank group, a clear and regular transmitted shadow was
observed. After 12 weeks, CT examination showed that the hydroxyapatite/gel was connected tightly with the surrounding normal bone tissue. As a new bionic composite, the hydroxyapatite/gel can achieve good effect in repairing skul defects of rabbits.

Background Whether negative suction during excimer laser in situ keratomileusis(LASIK) affect the structure and function of retina or not is in controversy,but it seems that temporary hypertention induced by negative suction is a key factor of impairment of retina in LASIK.ObjectiveThis study attempts to study the influence of transient high intraocular pressure (IOP) during LASIK on the contents of retinal amino acid.MethodsThe both eyes of 45 New Zealand white rabbits were suctioned for different periods (20s,45s,3min) with negative pressure generator during the LASIK to make the instantaneous high IOP models,and LASIK without negative suction was performed in the both eyes of 15 rabbits in the control group.The changes of the contents of retinal amino acids were evaluated with High Performance Liquid Chromatography(HPLC) at 0,7,10,14 and 28 days postoperatively and compared with those of control group.ResultsThere were no statistically significant differences in the contents of retinal amino acids among different time points after operation in negative suction for 20s group and 45s group,respectively(P＞0.05).At negative suction for 3minuts,the content of glutamic acid in retina was significantly increased in comparison with control group in 7,10,14 and 28 days (P＜0.05),and no statistically significant difference was seen in the contents of glutamic acid at postoperative instant group compared with control group(P＞0.05).A statistically significant difference in the contents of glutamine,tryptophan,phenylalanine was revealed among postoperative 10 days,14 days and 28 days groups comparison with control group (P＜0.05).ConclusionThe acute IOP elevation caused by negative suction during LASIK results in the reversible increase of retinal amino acids.The duration of negative suction time influent the reconstruction of retinal structure.

Objective:To establish an HPLC method to determine the content of carfilzomib for injection. Methods:The chroma-tographic column was a Waters symmetry C18(250 mm ×4.6 mm, 5 μm)column, the sample solvent was methanol, the mobile phase was 0. 05% trifluoroacetic acid-acetonitrile(57 ∶43) at a flow rate of 1. 0 ml · min-1 , the detection wavelength was 210 nm, the col-umn temperature was 25℃ and the injection volume was 10 ml. Results:The linear range of carfilzomib was 120-600 μg·ml-1 ( r=0. 999 7). The average recovery was 100. 4%( RSD=0. 24%,n=9). Conclusion:The method is rapid and effective, which can be used for the content determination of carfilzomib for injection.