Aim To observe the influence of carvedilol on the injury and expression of intercellular adhesion molecule-1 induced by hydrogen peroxide in ECV-304 cells and investigate the anti-atherosclerotic effect of carvedilol.Methods: The viability of ECV-304 cells was detected by MTT assay.Morphological changes of ECV-304 cells were observed under converse microscope.The level of lactate dehydrogenase released to the extracellular medium,the intracellular superoxide dismutase activity and the extracellular and intracellular Malondialdelyde level were determined using automatic biochemistry analyser.The expression of ICAM-1 in protein level and mRNA level was detected with flow cytometric technique and RT-PCR.Results Pretreated with carvidilol(1.0?10~(-5)～1.0?10~(-9)mol?L~(-1)) for 24 h,the cell survival rate was increased significantly in a concentration-dependent manner.Pre-incubation for 24 h with carvedilol results in a significant concentration-dependent decline of LDH release from hydrogen peroxide(1.0?10~(-6)mol?L~(-1))injured cells.While ECV-304 cells were pre-incubated with carvedilol,the level of MDA decreased and the activity of SOD increased significantly.Carvedilol produced a concentration-dependent inhibition of the expression of ICAM-1 protein and mRNA in hydrogen peroxide injured ECV-304 cells in a similar manner.Conclusion: These experiments demonstrated that carvedilol was able to protect ECV-304 cells from the oxidative stress injury and inhibit ICAM-1expression in ECV-304 cells induced by hydrogen peroxide.Therefore,we can consider that carvedilol maintains and improves the function of endothelium damaged by hydrogen peroxide from many aspects,which does indicate extensive antioxidant effects on the hydrogen peroxide-injured vascular endothelial cells and suggest promising effects in atherogenesis process.

OBJECTIVESTo evaluate the technical feasibility, safety, outcome, and incidence of complications after combined clear corneal phacoemulsification (PEA) with intraocular lens (IOL) implantation and vitreoretinal surgery.

METHODSCombined operations of PEA and PPV were performed on 52 eyes of 52 patients with cataract and vitreoretinal diseases.

RESULTSThe mean follow-up time was (10.3+/-2.8) months. Postoperatively, visual acuity improved in 46 eyes (88.5%); was unchanged in 6 eyes (11.5%). The best-corrected visual acuities (BCVAs) were the following: 20/40 or better (9 eyes), 20/50 to 20/100 (24 eyes), 20/200 (5 eyes), 20/400 (10 eyes), and fingers counting (FC) to light perception (LP) (4 eyes). In 38 eyes BCVA was 20/200 or better, and in 9 eyes it was 20/40 or better postoperatively. Postoperative complications included posterior capsual opacification (7 eyes); secondary glaucoma (1 eye); and retinal detachment (2 eyes).

CONCLUSIONAlthough further studies are indicated, our study suggests that the combined operation of PPV, PEA and IOL implantation is safe and effective for patients. The visual outcome and complications depended primarily on underlying posterior segment pathology and were not related to the combined procedure technique.

Adult ; Aged ; Cataract ; Female ; Humans ; Lens Implantation, Intraocular ; methods ; Lenses, Intraocular ; Light ; Male ; Middle Aged ; Perception ; Phacoemulsification ; methods ; Time Factors ; Treatment Outcome ; Vision, Ocular ; Visual Acuity ; Vitrectomy ; methods

To evaluate the phacoemulsification and intraocular lens implantation in patients with sensory exotropia subsequent to senile cataract. The authors prospectively studied the role of phacoemulsification and intraocular lens implantation on 25 patients by observing visual acuity, ocular alignment, binocular vision and diplopia pre-, 1 month post- and 3 months post-operation. The patients underwent follow-up for three months. Postoperatively, one patient had a corrected visual acuity of 20/50, and 24 patients had 20/40 or better. The ocular alignment, binocular vision and diplopia were resolved spontaneously. Phacoemulsification and intraocular lens implantation performed together is effective on sensory exotropia subsequent to senile cataract.
Aged ; Cataract ; complications ; diagnosis ; Combined Modality Therapy ; Exotropia ; diagnosis ; etiology ; surgery ; Humans ; Lens Implantation, Intraocular ; methods ; Phacoemulsification ; methods ; Recovery of Function ; Treatment Outcome ; Vision Disorders ; diagnosis ; prevention & control

In order to observe the effect of inhibitors for demethylation and histone deacetylase on the growth of leukemia cell line K562 and the expressin of tumor related genes, the K562 cells were treated with 5-aza-2' deoxycytidine (DAC) and trichostatin A (TSA) in co-culture; the growth curves were observed; the cell cycle was detected by flow cytometry (FCM); the gene expression pattern before and after drug treatment was measured with Atlas7742-1 microarray. The results showed that the combination treatment of DAC and TSA inhibited the proliferation of K562 cells, the growth of most cells were stopped in G(1)/S phases after drug treatment, the gene expression after treatment was more than before, and a few gene expression were down-regulated. In conclusion, combination treatment of DAC and TSA had an inhibitive effect on the leukemia cell line K562, combination of DAC and TSA with microarray could be used for screening candidate genes inhibiting leukemia cells.
Azacitidine ; analogs & derivatives ; pharmacology ; Cell Cycle ; drug effects ; Cell Division ; drug effects ; DNA Methylation ; drug effects ; Enzyme Inhibitors ; pharmacology ; Histone Deacetylase Inhibitors ; Humans ; Hydroxamic Acids ; pharmacology ; K562 Cells ; drug effects

OBJECTIVETo assess the efficacy of denaturing high-performance liquid chromatography (DHPLC) as a new screening assay for sequence variation in SNP detecting and new approach to SNP genotyping.

METHODSDHPLC was used to screen SNPs in 8 DNA pools each consisting of DNA from 5 individuals, and genotype the identified SNPs in 150 Chinese subjects from Hong Kong.

RESULTSSeventeen SNPs were identified: 12 were novel and 5 were previously reported; 11 were found in screening stage and the other 6 were found in genotyping stage; 2 were only found in Caucasian samples; 3 showed ethnic difference of minor allele frequency(MAF); 7 were common with the MAF>0.05 in Chinese samples.

CONCLUSIONThe results demonstrated the efficiency of DHPLC in screening SNPs when coupled with DNA pooling strategy, and in genotyping SNPs.

Alcohol Oxidoreductases ; genetics ; Chromatography, High Pressure Liquid ; methods ; Genotype ; Humans ; Point Mutation ; Polymorphism, Single Nucleotide ; genetics ; Sequence Analysis, DNA

The study was aimed to investigate the promotive effect of LRP16 gene on K562 cell proliferation. Open reading frame of LRP16 gene was amplified using reverse transcription-polymerase chain reaction (RT-PCR) and ligated to pGEM-T plasmid to construct LRP16 ORF-pGEM-T recombinant vector. Then, LRP16 ORF identified by sequencing was inserted into pcDNA3.1+ plasmid to construct LRP16 ORF-pcDNA3.1+ recombinant expression plasmid which was transfected into K562 cell lines to make overexpression of LRP16 gene in K562 cells. Survival of cells was determined by MTT assay and growth curve of cells was drawn, the cell cycle was detected by flow cytometry. The results showed that LRP16 ORF was successfully amplified, then the LRP16 ORF-pcDNA3.1+ recombinant plasmid was constructed. The K562 cell line with overexpression of LRP16 gene was established. The promotive effect of LRP16 gene overexpression on proliferation of K562 cells was observed and the effect partially related to the enhancement of cells from G0 to S phase induced by LRP16 gene. It is concluded that LRP16 gene overexpression shows a promotive effect on proliferation of K562 cells.
Cell Proliferation ; Genetic Vectors ; Humans ; K562 Cells ; Neoplasm Proteins ; genetics ; Open Reading Frames ; Plasmids

OBJECTIVETo investigate the effect of hyperbaric oxygen(HBO)therapy on mitochondrial free radicals after transient focal cerebral ischemia in rats.

METHODSThe male SD rats were randomly assigned into two groups, control and HBO groups. All animals were subjected to 90 min intra-luminal middle cerebral artery occlusion (MCAO) with the regional cerebral blood flow monitored in vivo by laser Doppler flowmetry. HBO treatment was performed in a pressure chamber with 100% O(2)(3 ATM 1 h) 3 h after ischemia. Twenty-four hours after ischemia, mitochondria in the ischemic core and penumbra were isolated and the contents of H(2)O(2), O(2)(*-), MDA, SOD, GSH-PX and GSH in mitochondria were measured respectively.

RESULTAfter cerebral ischemia-reperfusion, contents of mitochondrial H(2)O(2), O(2)(*-), MDA increased, while the SOD, GSH-PX and GSH in the mitochondria decreased significantly both in the ischemic core and the ischemic penumbra, compared with those in the normal controls(P<0.05). In the ischemic penumbra, HBO therapy increased significantly the content of O(2)(*-)(P<0.05), enhanced the activity of SOD, and decreased the level of MDA (P<0.05). However, HBO therapy did not change the level of MDA, though it also increased the content of O(2)(*-) and the activity of SOD in the ischemic core. HBO therapy had no significant effect on the contents of H(2)O(2), GSH-PX and GSH in the ischemic mitochondria.

CONCLUSIONHBO therapy initiated early after acute transient cerebral ischemia in rats can increase the mitochondrial free radicals level, but also increase the activity of the anti-radical enzymes. HBO treatment inhibits the lipid peroxidation damage of mitochondria in the ischemic penumbra, but not in the ischemic core, which indicates that the mitochondrial function plays a role in the reaction of the free radical in the ischemic area after HBO therapy.

Animals ; Free Radicals ; metabolism ; Glutathione Peroxidase ; metabolism ; Hyperbaric Oxygenation ; methods ; Infarction, Middle Cerebral Artery ; metabolism ; therapy ; Ischemic Attack, Transient ; metabolism ; therapy ; Male ; Mitochondria ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; prevention & control ; Superoxide Dismutase ; metabolism

OBJECTIVETo investigate the influence of location and size of acute insular infarct on stroke-related electrocardiogram (ECG) changes and cardiovascular events.

METHODSNinety-nine cases admitted to hospital from October 2007 to June 2009, who were diagnosed as acute middle cerebral artery territory infarct within 48 h after onset and without the history of cardiac diseases, were included in the study. The patients were further divided into three groups: major insular infarct, minor insular infarct and control group, according to the infarct size on MRI diffusion-weighted image. The clinical data, ECG changes and cardiovascular events were compared between left and right insular infarct. Logistic regression was applied to determine the independent risk factors of ECG changes and cardiovascular events.

RESULTLarge artery atherosclerosis was the main cause of acute insular infarct (71.8 %), which was associated with higher NIHSS score compared to the control group (P < 0.01). Comparing the left and right insular infarct, the frequencies of sinus bradycardia and sudden cardiac death were significantly higher in left insular infarct (P < 0.01 and P < 0.05), while there was a trend that the frequency of atrial fibrillation was higher in right insular infarct (P = 0.079). With the larger size of insular infarct, the frequency of sinus bradycardia, new atrial fibrillation and sudden cardiac death (P<0.01, P<0.05 and P<0.05, respectively) became much higher. Logistic regression analysis showed that major insular infarct was related to the higher frequency of sinus bradycardia (OR = 4.660, 95% CI: 1.646 ~ 13.195; P = 0.004).

CONCLUSIONAcute insular infarct is associated with the stroke-related ECG changes and sudden cardiac death. Left insular infarct is related to sinus bradycardia, possibly due to the enhanced parasympathetic tone. It deserves clinical attention that the incidence of cardiac autonomic disturbance becomes higher with the enlarged insular infarct size.

Acute Disease ; Adult ; Aged ; Aged, 80 and over ; Brain Infarction ; complications ; physiopathology ; Death, Sudden, Cardiac ; etiology ; Electrocardiography ; Female ; Humans ; Logistic Models ; Male ; Middle Aged ; Risk Factors

This study was purposed to investigate lrp16 gene expression in leukemia cell lines and bone marrow cells of leukemia patients and explore the relationship between lrp16 gene expression and development of leukemia. Reverse transcriptase-polymerase chain reaction (RT-PCR) was employed to test the lrp16 mRNA expression in 4 leukemia cell lines, including K562 (CML), HL-60 (APL), MOLT4 (ALL) and U937 cell lines, as well as in bone marrow-derived cells from 115 patients with leukemia. The effect of lrp16 gene expression on genesis and progression of leukemia was analyzed according to clinicopathological features. The results indicated that positive expression of lrp16 mRNA was found in all 4 leukemia cell lines. For leukemia patients, the positive expression rate of lrp16 mRNA in all AML patients was 38% (16/42), in which the positive rates in AML patients with complete remission (CR) and AML patients without remission were 13% (4/30) and 100% (12/12) respectively. The positive expression rate of lrp16 mRNA in ALL patients was 38% (10/26), in which the positive rate in ALL patients with CR and ALL patients without remission were 16% (3/18) and 87% (7/8) respectively. The positive expression rate of lrp16 mRNA in CML patients was 36% (9/25), in which the positive rates in CML patients with CR and CML patients without remission were 20% (4/20) and 100% (5/5) respectively. The positive rate of lrp16 mRNA in CLL patients was 31% (7/22), in which the positive rate in CLL patients with CR and CLL patients without remission were 11% (2/17) and 100% (5/5) respectively. There was no difference of lrp16 gene expression between leukemia subtypes, but there was statistical significant difference in lrp16 gene expression between CR patients and non CR patients (p < 0.001). It is concluded that the lrp16 gene is a leukemic oncogene and closely relates to genesis and progression of leukemia, which may be an indicator for evaluating clinical efficacy of leukemia therapy.
Adolescent ; Adult ; Aged ; Bone Marrow ; metabolism ; pathology ; Female ; HL-60 Cells ; Humans ; K562 Cells ; Leukemia ; metabolism ; pathology ; Male ; Middle Aged ; Neoplasm Proteins ; genetics ; metabolism ; RNA, Messenger ; genetics ; Young Adult

Background and Objective:LRP16 is a human novel gene linked to leukemia identified recently.However. its biological function is not fully clarified so far.This study was to investigate the biological function of human LRP16 gene by database-aided bioinformatics analysis. Methods:The structures and functions of LRP16 gene promoter and its coding protein were analyzed using bioinformatics prediction,and further experimental testing was performed.The recombinants of pGL3-basic and LRP16 promoter subclones were constructed for luciferase activity analysis.The recombinant of LRP16 open reading frame coding sequence and pcDNA3.1 eukaryotic expression vector was established and transfected into HL-60 and K562 cell lines.DNA damage of HL-60 cells after ultraviolet irradiation was evaluated using single cell gel electrophoresis. Cell cycle of K562 cells was analyzed by flow cytometry. Results:LRP16 promoter was a typical class Ⅱ eukaryotic promoter and its core regulation sequence was located within upstream -600 bp of transcriptional start site.In addition,seven cis-acting elements,which may be implicated in cell cycle,hematopoiesis regulation, cell proliferation and repair of DNA damage,were identified.Long type LRP16 coding protein contained homologous sequences of hismacro, COG2110,and A1PP with human histone H2A1C between 148 and 315 amino acid residue.The number of comet cells and the length of comet tail in HL-60 cells irradiated were significantly decreased and the number of living cell was significantly increased in LRP16-overexpression group compared with empty plasmid control group.The proliferation rate and ratio or quantity of G_2/M and S phases were significantly increased in LRP16-overexpression K562 group compared with empty plasmid control group.LRP16-overexpression in K562 cells promoted the transition of G_1 to S phase and plateau phase of cell proliferation was advanced.Conclusions:Promoter regulation prediction and protein domain analysis based on bioinformatics contribute to the study of gene function.LRP16 may play an important role in leukemia progression by promoting cell proliferation,regulating cell cycle,and antagonizing radiationinduced DNA damage.