Objective To observe the effect of intravitreal lucentis injection combined with macular grid photocoagulation for diabetic macular edema (DME).Methods 42 DME patients(42 eyes) according to different treatment methods,were divided into the combined group 21cases(21 eyes) and photocoagulation group 21 cases (21 eyes).The combined group was treated with connaught suitable for intravitreal injection combined macular grid photocoagulation , and the photocoagulation group was treated with simple macular grid photocoagulation .After 1 weeks,2 weeks,4 weeks,8 weeks,12 weeks and best corrected visual acuity before treatment (BCVA) and macular foveal thickness ( CMT) changes were comparative analyzed between the two groups .Results The BCVA values of the photocoagulation group before surgery ,1 week,2 weeks,4 weeks,8 weeks,12 weeks,were (0.74 ±0.24), (0.56 ±0.22),(0.52 ±0.23),(0.51 ±0.21),(0.51 ±0.20),(0.53 ±0.21),and CMT values were (440.36 ±50.56)μm,(376.06 ±28.60)μm,(365.45 ±30.02)μm,(361.38 ±29.56)μm,(358.68 ±30.46)μm,(360.43 ±29.36)μm;The BCVA values of the combined with preoperative group before surgery ,1 week, 2 weeks,4 weeks,8 weeks,12 weeks,were (0.70 ±0.23),(0.55 ±0.19),(0.52 ±0.18),(0.50 ±0.20), (0.48 ±0.19),(0.47 ±0.20),and CMT values were (442.32 ±54.35)μm,(385.14 ±27.34)μm,(345.32 ± 28.12)μm,(336.24 ±28.17)μm,(326.45 ±27.65)μm,(315.36 ±28.23)μm.The differences of BCVA and CMT values of the two groups in each time period values before and after treatment were statistically significant ( all P<0.05),and after 8 weeks,12 weeks,the differences of BCVA and CMT values between the two groups were statisti-cally significant(all P<0.05).Conclusion The macular grid photocoagulation only or lucentis injection combined with macular grid photocoagulation for diabetic macular could increase BCVA and reduce macular edema instinctively , but the difference was not significant .Combined therapy could preserve the visual acuity and delay time to relapse .

Objective To evaluate the application value of the blood culture combined with the detection of serum procalcitonin and C-reactive protein(CRP)in diagnosing infectious fever patients.Methods The PCT and CRP detection results on the blood sampling day in 126 cases of fever with positive blood culture and 123 cases of fever with negative blood culture in the emergency department were retrospectively analyzed,and the two sets of data were compared and statistically analyzed by the SPSS 16.0 soft-ware.Results The positive rates of CRP and PCT in 126 cases of positive blood culture were 100% and 100%,the detection values were 26.9-256.8 and 0.25-98.6,the positive rates of CRP and PCT in 123 cases of negative blood culture 95.5% and 15.4% re-spectively,the detection values were 1 .2-126.8 and 0.02-0.98.The PCT and CRP detection values in the positive blood culture group were significantly higher than those in the negative blood culture group,the difference was statistically significant(P <0.01). Conclusion PCT and CRP can be served as the monitoring indicators for blood bacterial infections,the blood culture combined with PCT and CRP detection has the guidance significance for early diagnosing bacterial infection.

Adolescent ; Child ; Child, Preschool ; Female ; Humans ; Infant ; Isocitrate Dehydrogenase ; genetics ; Leukemia, Myeloid, Acute ; genetics ; Male ; Mutation ; Nuclear Proteins ; genetics ; Prognosis ; Treatment Outcome

OBJECTIVEE23K polymorphism in KCNJ11 gene is associated with cardiovascular disease and diabetes. In order to explore the mechanism of E23K correlation to related diseases, the effect of E23K polymorphism in KCNJ11 gene on membrane current was investigated.

METHODSThe exon of KCNJ11 was obtained by PCR amplification and the G-->A mutation was completed by overlap extension PCR. The sequences of KCNJ11 exon contained 23E or 23K was inserted into pcDNA3.1/CT-GFP vector respectively. The recombinant plasmid, pcDNA3.1-KCNJ11(E) and pcDNA3.1-KCNJ11(K), were transfected into HEK293T cells by lipofectamine and the membrane current density was determined by whole-cell patch clamp technique.

RESULTS1,173 bp sequences of KCNJ11 gene's exon were amplified by PCR and the recombinant expression plasmid, pcDNA3.1-KCNJ11(E) and pcDNA3.1-KCNJ11(K), were constructed successful. Positive and negative currents were detected in HEK293T cells transfected with difference plasmid by whole-cell patch clamp technique. Results showed that the reversed voltage was 50mV. The current in HEK293T cells with pcDNA3.1-KCNJ11(E) was significantly greater than that with pcDNA3.1-KCNJ11(K) (P < 0.05, n = 10).

CONCLUSIONThe polymorphism of E23K in exon of KCNJ11 gene changed the membrane currents in HEK293T cells. It could be an experiment support for the possible mechanism between the locus and related diseases.

Objective To investigate the effect of intracoronary application of tirofiban on coronary slow flow patients with acute myocardial infarction during primary percutaneous coronary intervention (PPC1).Method It was a retrospective analysis of 187 patients with acute myocardial infarction treated with PPCI in the emergency department of Beijing Anzhen Hospital enrolled in this study from January,2008 through January,2011.The patients divided into 2 groups in terms of intra-coronary administration of tirofiban (tirofiban group) and intra-coronary use of nitroglycerol (control group).Data were statistically analyzed by using SPSS 13.0 software.Categorical variables were analyzed using x2 test and continuous variables were compared by t test.Results Between two groups,there were no differences in preoperative systolic pressure (P =0.245),the rate of TIMI flow 3 (P =0.568) after PPCI and ST segment resolution (P =0.824),LVEF (P =0.275) and in-hospital mortality (P =0.502).Compared with tirofiban group,the systolic pressure was lower and the rate of using intra-aortic counter-pulsation was higher in control group.Although the incidence of slight bleeding in the control group was lower than that in the tirofiban group,no severe bleeding was observed in both groups.Conclusions The effect of intracoronary use of tirofiban was similar to that of nitroglycerol in terms of improving slow flow of coronary artery.It could safely and effectively reduce the incidence of the coronary slow flow in the patients after PPCI,but it produced a little impact on systolic pressure.It may be a better method of choice for AMI patient with low blood pressure.

Objective To investigate the role of nucleotide-binding oligomerization domain-containing protein 1 (NOD1) in inducing insulin resistance in differentiated human adipocytes.Methods Human preadipocytes obtained via liposuction were induced to differentiate into mature adipocytes.iE-DAP,a specific ligand for NOD1,was administered to human adipocytes in culture.NF-κB transcriptional activity and proinflammatory cytokines production were determined by luciferase assay and enzyme-linked immunosorbent assay.Glucose uptake in adipocytes was measured by 2-deoxy-D-[3 H] glucose uptake (P＜0.05).The expression of phosphatidylinositol-3-kinase p85 (PI-3K p85),insulin receptor substrate-1 (IRS-1) and Akt phosphorylations were detected by Western blotting.Results NF-κB transcriptional activity and cytokines such as interleukin (IL)-6,IL-8,and monocyte chemotactic protein 1 secretion were markedly increased after stimulation with iE-DAP (P＜0.05 or P＜0.01).Insulin-induced glucose uptake was decreased with the activation of NOD1 in a dose-and time-dependent fashion.NOD1 activation weakened insulin signal transduction as being revealed by increasing IRS-1 Ser307phosphorylation,reducing protein expression of PI-3K p85,and attenuating insulin-induced phosphorylation of Akt on Ser473 and Thr308in human adipocytes.Conclusion These results indicate that NOD1 activation induces inflammatory response and insulin resistance via IRS-1/PI3-K/Akt signaling pathway in human adipocytes.

Aim To investigate the effects of Aplysin on the inhibition of gastric cancer cell in vitro .Methods MTT assay was used to examine the inhibition of gastric cancer cell 1ine SGC-7901 by Aplysin in different concentrations and at different times.The morphologic changes and the apoptosis of SGC-7901 was observed by inverted microscope and Hematoxylin-Eosin(HE)staining.Reverse transcriptase polymerase chain reaction(RT-PCR)assay was used to detect the changes of COX-2 mRNA expressions.Results Aplysin could decrease the proliferation significantly in a dose-dependent manner in SGC-7901 cells.When treating SGC-7901 with Aplysin in concentration of 120, 240 mg·L~(-1) for 24 h, the growth of the cell was obviously inhibited observing by inverted microscope.Aiso, when treating with the same concentration for 18 h, its chromatin became crimpled and breakdown, as well as cell shrinkage and apoptotic bodies formation when using HE staining.The apoptotic rates(%)of SGC-7901 was(15.0±2.12)%, (18.4±2.3)%, respectively, which was significantly higher than(1.4±0.55)% that in control group(P <0.01).60、120、240 mg·L~(-1) Aplysin could not effectively inhibited the mRNA expressions of COX-2(P ＞0.05).Conclusions Aplysin can inhibit the proliferation and induces apoptosis of SGC-7901 cells.

Objective To observe the abnormality on behavioral ability of transgenic mice for HRX-EEN fusion gene.Methods Transgenic mice for HRX-EEN fusion gene(transgenic group,n=12)and C57/BL mice(control group,n=12)were tested in hidden platform training(day 1 to day 4)and probe trial testing(day 5)in Morris water maze in which ability of spatial learning and retention was assessed.Results In hidden platform training,the latencies of transgenic group were longer than those in control group,and significant differences were observed between the two groups for day 2,3 and 4(P

Objectiive:The present study was designed to explore whether overexpression of human wild ?-synuclein in rat brain caused selective dopaminergic neuron loss in substantia nigra and aimed to find out a new method to make a rat model of Parkinson's disease(PD).Methods:The human wild ?-synuclein gene was induced into the rat brain by Adeno-Associated Virus(AAV) vector.The overexpression of ?-synuclein was detected by realtime PCR.The behavior of rats were recorded every 4 weeks after the viral particle injection.TH immunohistochemistry were performed at 4,8,12 and 16 weeks post-injection as well as the dopamine(DA),3,4-dihydroxypheny-lacetic acid(DOPAC) of striatum were determined by high performance liquid chromatography coupled with electrochemical detection.Results:Realtime PCR results revealed a significant overexpression of ?-synuclein in the injected hemisphere.By 8 weeks post injection,a significant loss of the dopaminergic neurons was observed.34% of the dopaminergic neurons were lost after 12 weeks,and about 60% cells loss after 16 weeks.The DA and DOPAC levels in the striatum decreased about 15% 12 weeks after injecting viral particle carried ?-synuclein gene and 30% decreased after 16 weeks.The AAV-?-synuclein-treated rats developed a type of motor impairment,i.e.,head position bias,compatible with this magnitude of nigrostriatal damage.Conclusion:All the results showed that overexpression of human wild ?-synuclein caused selective dopaminergic neuron loss and mimic a symptom of human PD in rats.This may be a new methed to make rat PD model which can offer new opportunities for the study of pathogenetic mechanismsand exploration of new therapeutic targets of particular relevance to human PD.

Aim To investigate the protective effects of Aplysin on ethanol-induced oxidative damage in rat pri-mary hepatocytes. Methods Rat primary hepatocytes were obtained via the portal vein collagenaseⅣin situ perfusion technique followed by a Percoll density gradi-ent centrifuge. MTT test was used to determine the op-timum dose of Aplysin and ethanol, and detect the cell vitality in primary hepatocytes. Supernatants of primary hepatocytes were harvested to measure AST and LDH level, and the SOD, GSH-PX activities and MDA con-tent in primary hepatocytes were observed. Flow cy-tometry was used to detect the cell apoptosis rate. DNA damage in primary hepatocytes was detected by single-cell gel electrophoresis assay. The level of mitochon-drial membrane potential in primary hepatocytes was tested by fluorogenic probe JC-1 . The CYP2 E1 activity in primary hepatocytes was detected by colorimetry. The proteins of CYP2 E1 were detected by Western blot. Results 300 mmol·L-1 dose of ethanol and 30 mg·L-1 dose of Aplysin were the optimal dosages and were used in the subsequent experiments. Hepatocyte vitality was significantly increased in Aplysin group compared to that in ethanol group, and Aplysin inhibi-ted the release of AST and LDH(P<0. 05). For Apl-ysin treatment group, the activities of hepatocyte SOD and GSH were significantly increased, and MDA was markedly lowered as compared with those in ethanol group( P <0. 05 ) . Aplysin could alleviate hepatocyte apoptosis significantly, and hepatocyte DNA damage rates of Ⅱ ~Ⅲ level and Ⅳ level were significantly lowered in Aplysin treatment group as compared with those in ethanol group, and Aplysin had evident im-provement in alcohol induced mitochondria damage of hepatocyte. Primary hepatocyte activities and protein expression of CYP2 E1 were markedly lowered in Aply-sin treatment group as compared with those in ethanol group(P<0. 05). Conclusion Aplysin has protective effects on liver oxidative damage induced by alcohol of primary cultured rat hepatocytes by blocking CYP2 E1 activation, relieving oxidative stress, and sharpening the oxidation resistance ability.