BackgroundProliferative vitreo-retinal disease (PVD)is one group of ocular complications marked by the enhanced proliferation of various cells included retinal pigment epithelial (RPE) cells.Insulin-like growth factor-1 (IGF-1)and vascular endothelial growth factor(VEGF) are implicated in the aberrant cell proliferation and pathological neovascularization that characterizes PVD,but the signaling mechanism is unclear now. Objective This study was to explore the effect of IGF-1 on VEGF in cultured human RPE cells under the small hairpin loop RNA (shRNA) keeping hypoxia-inducible factor-1 α ( HIF-1 α) silencing. Methods Human retinas were isolated from 4 healthy male donors,and the RPE cells were harvested and cultured.The ceils were identified using anti-human keratin antibody.The third to fifth generation of human RPE cells were used in the experiment.One target site of HIF-1α mRNA was chosen by certain design principle,and shRNA was designed and synthesized by the target site and transferred into the cells in vitro,and then the cells were cultivated with 50 μg/L IGF-1 for 24 hours.The mRNA and protein expressions of HIF-1α and VEGF were detected by RT-PCR and Western blot respectively. Results Cultured human RPE cells showed the flat irregularly multangular shape,and 97％cells appeared the positive response for keratin.HIF-1α mRNA expression in human RPE cells was significantly lower in 50 μg/L IGF-1 group than the 0 pg/L IGF-1 group ( 1.49±0.18 vs 1.46±0.17 ) ( t =0.335,P =0.743 ),however,the expressing levels of HIF-1α protein( 1049.86±172.54 vs 0.00±0.00) and VEGF mRNA(0.95±0.15 vs 0.35±0.07) and VEGF protein (391.98±56.77 vs 214.36±37.15)were raised in the 50 μg/L IGF-I group compared with 0 μg/L IGF-1 group (t=16.098,9.935,6.928,P＜0.05).After the HIF-1α-specific shRNA was transferred into cultured RPE cells,the expressions of both HIF-1α mRNA and its protein significantly decreased in RPE cells under 50 μg/L IGF-1 concentration condition( F=68.679,89.904,P=0.000),moreover,the expression of VEGF mRNA and its protein were significantly lowed(F=21.770,6.205,P＜0.05). ConclusionsIGF-1 promotes the accumulation of HIF-1α protein and induce the expression of VEGF in human RPE cells,which probably play a pivotal role in the development of PVD.

Objective To investigate the difference in influerce of complicated diabetes on the prognosis of kidney transplantation.Methods Analyze the data of 507 times kidney transplantation and clinical tracing.Results Totally 507 patients with kidney function failure had received 539 times kidney transplantation,52 of whom were connected with diabetes[30 male,(51.4?7.1)age;22 female,(51.6?4.9)age],and 16 cases were pre-transplant,36 cases post-transplant.The other 455 cases were non-diabetes[293 male,(38.7?12.3)age;126 female,(43.5?10.1)age].The prevalence of diversified complications in the group with diabetes was all remarkably higher than the other group.There were 9 cases death in the group with diabetes,and 35 deaths in the other group after kidney transplantation;actually the mortality of diabetes group(17.3%,9/52)was 2.45 times as high as the group without diabetes(7.7%,35/455),especially in pre-transplant diabetes group.Another result was that there was no significant difference in the survival period of the dead patients in group with and without diabetes.Conclusion The possibility for the patients who,suffer from kidney function failure with diabetes to get Kidney transplantation postoperative complications is bigger compared with those without diabetes,and the prognosis is poor.To control diabetes more effectively before and after kidney transplantation perhaps can improve the prognosis of them.

Objective To observe the age-related changes of brain-derived neurotrophic factor(BDNF) and investigate the effects of ginsenosides(GS) on BDNF in nucleus basalis of Meynert(NBM) and cerebral cortex of aged rats. Methods Twenty-four female Wistar rats were randomly divided into 3 groups: young group(3-5 months),aged groups(27 months) and GS group(27 months).GS group was fed with GS from 18 to 27 months.Immunohistochemistry and Imaging analysis were used to show the expression and distribution of BDNF in NBM and cerebral cortex of each group. Results BDNF level of aged group was much lower than that of young group in these two brain areas(P

INTRODUCTIONErythroderma is a generalised inflammatory reaction of the skin secondary to a variety of causes. This retrospective study aims to characterise the features of erythroderma and identify the associated causes of this condition in our population.

MATERIALS AND METHODSWe reviewed the clinical, laboratory, histological and other disease-specific investigations of 225 inpatients and outpatients with erythroderma over a 7.5-year period between January 2005 and June 2012.

RESULTSThe most common causative factors were underlying dermatoses (68.9%), idiopathic causes (14.2%), drug reactions (10.7%), and malignancies (4.0%). When drugs and underlying dermatoses were excluded, malignancy-associated cases constituted 19.6% of the cases. Fifty-five percent of malignancies were solid-organ malignancies, which is much higher than those previously reported (0.0% to 25%). Endogenous eczema was the most common dermatoses (69.0%), while traditional medications (20.8%) and anti-tuberculous medications (16.7%) were commonly implicated drugs. In patients with cutaneous T-cell lymphoma (CTCL), skin biopsy was suggestive or diagnostic in all cases. A total of 52.4% of patients with drug-related erythroderma had eosinophilia on skin biopsy. Electrolyte abnormalities and renal impairment were seen in 26.2% and 16.9% of patients respectively. Relapse rate at 1-year was 17.8%, with no associated mortality.

CONCLUSIONOur study highlights the significant proportion of malignancy-related erythroderma in those whom common underlying causes such as dermatoses and drugs have been excluded. In cases of drug-related erythroderma, traditional medications and antituberculous medications are common causes in our population. Renal impairment and electrolyte abnormalities are commonly seen and should be monitored in patients with erythroderma.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Child ; Child, Preschool ; Dermatitis, Exfoliative ; diagnosis ; etiology ; Female ; Humans ; Infant ; Male ; Middle Aged ; Retrospective Studies ; Young Adult

Carcinoma, Squamous Cell ; diagnosis ; surgery ; Diagnosis, Differential ; Eye Evisceration ; Humans ; Male ; Middle Aged ; Orbital Neoplasms ; diagnosis ; surgery ; Ulcer ; diagnosis

Atomic force microscopy (AFM), as a sophisticated imaging tool with nanoscale resolution, is widely used in virus research and the application of functional viral particles. To investigate single viruses by AFM in a physiologically relevant environment (liquid), an appropriate surface treatment to properly adhere the viruses to the substrate is essential. Here we discuss hydrophobic treated glass coverslips as a suitable substrate for the adhesion of single adenovirus particle (Adenovirus type 5 F35, Ad5F35) when studied with AFM in liquid. From the high resolution AFM images, the orientation of the adhered virus particles can be distinguished. Furthermore, the particles exhibit the expected height of -90 nm. This illustrates that the viruses adhere to the substrate firmly without large deformations. Hence, the described method works well on (fragile) viruses. The described experimental approach can be widely used for AFM studies in liquid of virus structure and mechanics as well as for investigating the interaction of viruses with cellular receptors.
Adenoviridae ; chemistry ; Imaging, Three-Dimensional ; Microscopy, Atomic Force ; methods ; Virion ; chemistry

AIM:To investigate the protective effect of losartan(Los) on apoptosis of H9c2 cells induced by isoprenaline(ISO),and to discover its related mechanism.METHODS:H9c2 cells cultured on plastic plates were divided into control,ISO,ISO+Los,ISO+Los+LY294002 and DMSO groups.Cell apoptosis was evaluated by flow cytometery and agarose gel electrophoresis.The mRNA levels of bax,bcl-2 and caspase-9 were detected by RT-PCR and the expressions of phosphorylated and total Akt(p-Akt and t-Akt) were assessed by Western blotting.The cAMP was measured by radioimmunoassay.RESULTS:ISO at concentration of 10 ?mol/L induced apoptosis of H9c2 with an increase in bax/bcl-2,caspase-9 and cAMP.Addition of 10 ?mol/L losartan inhibited apoptosis obviously with a decrease in bax/bcl-2,caspase-9 and cAMP.A significant increase in p-Akt was observed,and its protein level was elevated.LY294002 at concentration of 1 ?mol/L abolished the protective effects of losartan on ISO-induced apoptosis in H9c2 cells.CONCLUSION:ISO might induce H9c2 cell apoptosis through stimulation of ?-adrenergic receptor(?-AR).Los inhibits downstream signaling of ?-AR,and promotes the activation of Akt.Subsequently it might attenuate the apoptosis induced by ?-adrenergic stimulation of ISO.

AIM: To study effects of urokinase-type plasminogen activator (uPA) signal transduction on expression of matrix metalloproteinase-2 (MMP-2) and tissue inhibitor of matrix metalloproteinase-3 (TIMP-3) in giant cell tumor of bone (GCT). METHODS: Expression of uPAR, MMP-2 and TIMP-3 in GCT tissue was detected by immunohistochemistry. Phosphorylation level of mitogen-activated protein kinase (p44) in uPA/uPAR signal pathway in cultured GCT cells was detected by immunoprecipitation. The expression of MMP-2 and TIMP-3 in cultured cells after treatment with uPA-ATF or anti-uPAR antibody was also detected by Western blotting. RESULTS: 1) Urokinase-type plasminogen activator receptor (uPAR) was positive on the cell membrane and in cytoplasm of some mononuclear stromal cells (MSCs) and multinucleated giant cells (MGCs); 2) MMP-2 was positive in the cytoplasm and on the cell membrane of almost all of MSCs and some of MGCs. The polar distribution of MMP-2 in the cytoplasm of MGCs was especially obvious; 3) The expression of TIMP-3 of some MSCs and MGCs in GCT was much lower than MMP-2. The positive signal also showed a prominent polarity; 4) After treatment with uPA-ATF, the phosphorylation level of p44 in GCT cultured cells was much higher than the control. Addition of anti-uPAR antibody in the cells remarkably down-regulated the phosphorylation level of p44 as compared with the control group, suggesting that uPA-ATF participates cell signal transduction and this reaction can be inhibited by anti-uPAR antibody; 5) uPA-ATF cell signal pathway up-regulated expression of MMP-2 and TIMP-3, while anti-uPAR antibody down-regulated the expression of MMP-2 and TIMP-3. CONCLUSION: These results demonstrate for the first time that uPA-ATF directly regulates the expression of MMP-2 and TIMP-3 by signal transduction pathway, and the over-expression of MMP-2 and TIMP-3 may play an important role in local osteolysis of GCT. [

Objective To observe the effect of pentylenetetrazole(PTZ)-kindled epliepsy on rat's learning and memory and the expression of calmodulin dependent protein kinaseⅡ(CaMKⅡ)in hippocampal neurons.Methods The rats were injected intraperitoneally with PTZ to establish the kindled models.The changes of behavior were evaluated through Morris water maze and the expression of ?-CaMKⅡ in hippocampal neurons was observed by immunohistochemistry technique.Results The Morris water maze test showed that the escape latency and the swimming distance in PTZ-kindled rats increased significantly(P

ObjectiveTo evaluate the functional changes of stunned myocardium before and after coronary artery bypass graft(CABG) treatment,and clear the meaning of revascularization which CABG has brought to patients with diffused vascular changes.MethodsA total of 36 patients with 99％ diffused coronary artery stenosis in left anterior descending branch underwent non-pump CABG treatment in the Department of Cardiothoracic Surgery the First affiliated hospital of Harbin Medical University.Real-time three-dimensional echocardiography (RT3DE) was repeatedly performed 1 week before operation and 10 days,1 month,and 3 months after CABG.Regional diastolic volumes,systolic volumes,ejection fractions,regional stroke volume to global diastolic volume and the values of abnormal segments before and after CABG were studied.ResultsOne week before operation and 10 days,1 month and 3 months after CABG,the differences of volumes between groups in the last phases of diastole and systolic were statistically significant in anterior wall basement segment,anterior septal basement segment,anterior wall intercalary segment,anterior septal intercalary segment,anterior wall of apex cordis and septation of apex cordis(F =3.51,3.55,4.08,4.05,2.98,3.01,all P ＜ 0.05; F =4.51,4.55,4.08,3.00,2.96,2.99,all P ＜ 0.05).The values of the six segments mentioned above,3 months after operation[(6.74 ± 1.23),(6.64 ± 1.21),(6.02 ± 1.10),(5.95 ± 1.09),(5.82 ± 1.06),(5.10 ± 0.93)ml; (2.74 ± 0.50),(2.69 ± 0.49),(2.51 ± 0.46),(2.32 ± 0.42),(2.36 ± 0.43),(2.03 ± 0.37)ml] were compared with those of 1 week before operation[(8.33 ± 1.52),(8.20 ± 1.50),(7.43 ± 1.36),(7.36 ± 1.34),(7.19 ± 1.31),(6.29 ± 1.15)ml; (4.94 ± 0.90),(4.85 ± 0.88),(4.53 ± 0.83),(4.18 ± 0.76),(4.25 ± 0.78 ),(3.65 ± 0.67)ml],the differences were statistically significant (all P ＜ 0.05); the differences between groups in regional ejection fractions,regional-global ejection fractions were statistically significant(F =4.56,4.88,4.28,3.15,2.93,2.88,P ＜ 0.01 or ＜ 0.05; F =5.56,5.28,4.98,5.15,3.03,2.78,P ＜ 0.01 or ＜ 0.05).Compared with 1 week before the operation, 1 month after the operation in regional ejection fractions,10 days,1 month in regionalglobal ejection fractions after the operation,4 segments of them were significantly improved(all P ＜ 0.05) and 3 months after operation,all the 6 segments had been improved significantly(all P ＜ 0.05).The maximum volume of the sum of group difference of the 6 segments and the 4 segments in the last phase of diastole was statistically significant(F =2.58,5.81,P ＜ 0.05 or ＜ 0.01 ),and the summation began to decrease 10 days after the operation.The values of 3 months after operation[ (36.27 ± 1.10),(25.35 ± 1.16)ml] were compared with that of 1 week before operation[ (44.80 ± 1.36),(31.32 ± 1.43)ml ] the difference was statistically significant (all P＜ 0.05).The maximum volume summafion comparisons of 6 segments and 4 segments in the last phase of systolic had statistical significance(F =5.77,5.57,all P ＜ 0.01 ),and 10 days after the operation,the summation began to decrease.The values of 1 month[(16.4 0 ± 0.48),(11.58 ±0.51 )ml],and 3 months after operation[ (14.65 ± 0.45),(10.26 ± 0.46)ml],were compared with those of 1 week before operation[ (26.40 ± 0.80),(18.50 ± 0.84)ml],the differences were statistically significant (all P ＜ 0.05).ConclusionsStunned myocardium can be improved through CABG in myocardium systolic,diastole function and ejection fractions of the relevant segments and all of this have proved that patients undergoing CABG revascularization can improve the heart function of the ischemic area.