Background The special pathological change of diabetic retinopathy(DR)is microvascular disorder.Angiopoietin-like protein 2(Ang-2)is a new protein associated with genesis of blood vessels.Quercetin has multiple pharmacological action,including improving the microcircularion and the permeability of blood capillary.However,the action mechanism of Ang-2 on DR was unclear.Objective The present study was to investigate the effects of quercetin on Ang-2 and its receptor Tie2 expression in retina with diabetes mellitus.Methods Sixty clean male Wistar rats were randomized into 7 groups and 10 rats for each group,and 10 rats served as blank control group.Streptozotocin of 35 mg/kg was intraperitoneally injected in 60 rats to establish the diabetic models.Quercetins encapsulated by liposome with the doses of 50,150 and 250 mg/(kg · d)(3-5 ml)were used to gavage in different groups of models for 12 weeks,and normal saline solution and calcium dobesilate were used at the same fashion as the negative control group and positive control group,respectively.Twelve weeks later,the animals were sacrificed and retinas were isolated.Expressions of Ang-2 protein and Tie2 mRNA in retinas were detected by ELISA and RT-PCR,respectively.The usage and rearing of the animals complied with the Regulations for the Administration of Affair Concerning Experimental Animals by State Sciences and Technology Commission.Results ELISA showed that the A450 of Ang-2 in 150 and 250 mg/(kg · d)quercetin groups was 0.796±0.057 and 0.842±0.043 respectively and was lower than that negative group(1.012±0.046),showing statistically significant differences(q =2.95,2.698,P＜0.05).RT-PCR assay showed that expression of Tie2 mRNA(Tie2 mRNA/GAPDH mRNA)in retinas was 0.712±0.092 and 0.821±0.087,presenting statistically significant differences in comparison with negative group(1.182±0.098)(q =3.497,2.852,P＜0.05).The expression levels of Ang-2 and Tie2 mRNA in retina were lowest in 150 mg/(kg · d)quercetin group.Conclusions Quercetin can improve the retinal microcirculation by downregulating the expressions of Ang-2 and its receptor in early period of diabetic rats.

Objective To evaluate the effect of off-pump coronary endarterectomy (CE) plus off-pump coronary artery bypass grafting (off-pump CABG) on patients with chronic total occlusion (CTO) combined with diffuse distal atherosclerosis. Methods From October 2006 to August 2009,65 CTO patients with 176 angiographically confirmed vascular stenosis or occlusive lesions, 70 of which were complete occlusion, underwent off-pump CABG. During the operation, diffuse intimal thickening distal to occlusive lesion was found, and blood flow of the bridges was unfavorable.Results Therefore endarterectomy was performed, followed by CABG. The blood flow in the bridges were 2-10 ml/min versus 14-37 ml/min before versus after endarterectomy. Pulsatility index (PI) was 5.1-15.6 versus less than 5 before versus after endarterectomy. Left ventricular ejection fraction was also improved significantly [before operation: (0.47±0.12)%, after operation: (0. 52±0.15)%, t=2.17, P＜0.05]. Peri-operative myocardial infarction occurred in 2 cases, but without significant cardiac homodynamic changes. And 23 patients underwent coronary angiography to evaluate graft patency 3-18 months after operation, all of them had favorable blood flow. Conclusions It is feasible to perform off-pump CABG plus coronary endarterectomy for patients of chronic coronary total occlusion combined with diffuse distal atherosclerosis. This treatment is safe and effective.

Objective To explore the effect of uhrasound-phoresis of Chinese herb in the treatment of frac- ture healing on rats.Methods The animal model of femur fracture was established in 36 rats,who were divided in- to 4 groups after operation.The rats in the ultrasound group were given ultrasound treatment daily.The herbal group was given Chinese herb applied on the fracture site.The experimental group was given uhrasound-phoresis of Chinese herb on the site of fracture.The control group was housed without any treatment.All rats were sacrificed at 30 days and the bony callus were harvested and observed with histological anti immunohistoehemical examination.Results The histological examination showed that the appearance of cartilaginous and bony callus in the experimental group were earlier than those in other groups,Immunohistochemical examination showed that the expression of collagenⅠin the experimental group was significantly higher than that in ultrasound group and herbal group(P

OBJECTIVETo investigate the expression status of 11 different cancer/testis (CT) antigen genes in esophageal carcinoma.

METHODSEsophageal carcinoma tissue and adjacent normal esophageal mucosa taken from 35 esophageal carcinoma patients were assayed for the expression of 11 different CT antigen genes by RT-PCR techniques.

RESULTSOf the 11 CT antigen genes analyzed, none of them was expressed in normal esophageal mucosa. MAGE-3 was found to be the most frequently expressed in esophageal carcinoma tissues (62.9%), followed, in the order of expression frequency, by MAGE4 (31.4%), LAGE-1 (28.6%), MAGE-1 (25.7%), CT10 (20.0%), NY-ESO-1 (20.0%), CT7 (5.7%) and SCP1 (2.9%). No expression of SSX-1, SSX-2 and SSX-4 was found. Among the 35 cases, 28 (80.0%) expressed at least one CT antigen gene, 21 (60.0%) expressed more than 2 CT antigen genes, and 4 of the 21 (19.0%) expressed more than 4 CT antigens, which accounted for 11.4% of total number of patients (4/35). No CT antigen expression was found in the tumor tissue in 7 cases, including 5 cases in stage II and 1 case each in stage I and IV, respectively. Of the 11 CT genes examined, expression of 5 genes (NY-ESO-1, LAGE-1, MAGE-1, MAGE-3 and MAGE-4) was correlated with tumor progression. SCP-1 and CT10 expression was found more frequently in early stage patients. With progression of the disease, the frequency of co-expression of multiple CT antigen genes was significantly increased reaching 28.6% in stage III patients.

CONCLUSIONOf the 11 different CT antigen genes examined by RT-PCR in esophageal carcinoma, 8 genes were detected in various frequencies in 28 of the 35 esophageal cancer patients studied. They are candidate tumor-associated antigens in the preparation of tumor vaccines for immunotherapy in esophageal cancer patients.

Adult ; Aged ; Antigens, Neoplasm ; biosynthesis ; genetics ; Base Sequence ; Esophageal Neoplasms ; genetics ; metabolism ; Female ; Humans ; Male ; Middle Aged ; Molecular Sequence Data ; Neoplasm Proteins ; biosynthesis ; genetics

BACKGROUNDThis study was undertaken to obtain differentially expressed genes related to human glioma by cDNA microarray and the characterization of a novel full-length gene.

METHODSTotal RNA was extracted form human glioma and normal brain tissue, and mRNA was used as a probe. The results of hybridization procedure were scanned with the computer system. The gene named 507E08 cone was subsequently analyzed by northern blot, bioinformatic approach, and protein expression.

RESULTSFifteen differentially expressed genes were obtained from human glioma by hybridization and scanning for four times. Northern blot analysis confirmed that the 507E08 clone was low expressed in human brain tissue and over expressed in human glioma tissues. The analysis of BLASTn and BLASTx showed that the 507E08 clone was a novel full-length gene, which codes 203 amino acid of protein and is called human ribosomal protein 14.22 gene. The nucleotide sequence had been submitted to the GenBank with the accession number of AF329277. After expression in E. coli., protein yielded a major band of apparent molecular mass 22 kDa on an SDS-PAGE gel.

CONCLUSIONScDNA microarray technology can be successfully used to identify differentially expressed genes. The novel full-length gene of human ribosomal protein 13.22 may be correlated with the development of human glioma.

Amino Acid Sequence ; Base Sequence ; Blotting, Northern ; Cloning, Molecular ; DNA, Complementary ; chemistry ; genetics ; Electrophoresis, Polyacrylamide Gel ; Escherichia coli ; genetics ; Glioma ; genetics ; pathology ; Humans ; Molecular Sequence Data ; Oligonucleotide Array Sequence Analysis ; RNA, Messenger ; genetics ; metabolism ; Recombinant Proteins ; isolation & purification ; metabolism ; Ribosomal Proteins ; genetics ; metabolism ; Sequence Analysis, DNA

OBJECTIVE: The STR genotypping of trace oral epithelial cells which are microdissected by laser capture microdissection system (LCM) is explored. METHODS: The oral epithelial cells are microdissected using a low-power infrared laser by VERITAS Microdissection Instrument. STR loci of Profiler Plus are detected by multiplex PCR procesures. RESULTS: DNA genotyping of 7-8 oral epithelial cells are succeeded, and DNA genotyping of 3-4 oral epithelial cells are failed. CONCLUSION It is viable in genotyping of trace oral epithelial cells by Laser Capture Microdissection as a new technology of seperating single cell.
Cell Separation/methods* ; DNA/genetics* ; Epithelial Cells ; Genotype ; Humans ; Lasers ; Microdissection/methods* ; Mouth/cytology* ; Tandem Repeat Sequences

OBJECTIVETo study the effect of exogenous caveolin-1 on the growth of laryngeal squamous cell carcinoma and its mechanisms.

METHODSEukaryotic expression vectors containing human caveolin-1 gene were transfected into Hep-2 cell line, the positive clones with high expression of caveolin-1 were identified by fluorescence quantitative real time reverse transcriptase-polymerase chain reaction and Western Blot. Cell proliferation viability was tested by methyl thiazolyl tetrazolium assay, the protein expression of epidermal growth factor receptor (EGFR), P-EGFR, extracellular signal-regulated kinase 1, 2 (Erk1, 2), P-Erkl, 2 and caveolin-1 were detected by Western Blot. The combination of caveolin-1 and EGFR were studied by immunoprecipitation and Western Blot. The in vivo antitumor activity of caveolin-1 was tested in Hep-2 xenograft tumor models in athymic nude mice, and the protein expressions of P-EGFR, P-Erk1, 2 and caveolin-1 were examined by immunohistochemistry.

RESULTSThree of caveolin-1 stably transfected Hep-2 cell clones were established. MTT assay showed that the proliferation of caveolin-1 overexpression Hep-2 cell clones decreased significantly comparing with the control. Immunoprecipitation and western Blot showed that caveolin-1 and EGFR were combined in Hep-2 cell line. Comparing with the parental cell line and cells transfected with control vector, there were the lower phosphorylation of EGFR and Erk1, 2 in the caveolin-1 overexpression Hep-2 cell clones. In the xenograft tumor models in nude mice, caveolin-1 overexpression Hep-2 cell clones showed the slower growth, smaller tumor size and the lower phosphorylation of EGFR and Erk1, 2.

CONCLUSIONSOverexpression of caveolin-1 inhibits growth of Hep-2 cell line in vitro and in vivo, arresting EGFR-MAPK signal pathway may involve in its mechanism.

Animals ; Carcinoma, Squamous Cell ; metabolism ; Caveolin 1 ; pharmacology ; Cell Line, Tumor ; Humans ; Laryngeal Neoplasms ; metabolism ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Mitogen-Activated Protein Kinase Kinases ; metabolism ; Receptor, Epidermal Growth Factor ; metabolism ; Signal Transduction ; Transfection

OBJECTIVETo investigate the effects of caveolin-1 on the biologic behavior of laryngeal squamous cell carcinoma HEp2 cell line in vitro.

METHODSEukaryotic expression vector of human caveolin-1 gene was constructed and transfected into HEp2 cells by Lipofectamine. The clones stably overexpressing caveolin-1 were identified by real-time PCR and Western blotting. Cell proliferation viability was tested by MTT assay. Anchorage-independent growth was determined by assaying colony formation in soft agar. Flow cytometry was used to assess the cell cycle and apoptosis. The relative phosphorylation level of EGFR and ERK1/2 were detected by Western blotting. Localization of caveolin-1 and EGFR were studied by laser confocal laser scanning microscopy.

RESULTSThe expression vector of caveolin-1 was constructed and three clones stably overexpressing caveolin-1 were obtained. Comparing with the parental HEp2 cells, the transfected cells exhibited a slower growth rate and formed fewer colonies in soft agar. The results of FACS analysis revealed that overexpression of caveolin-1 resulted in the cell cycle arrest at G0/G1 phase and increased the apoptotic cell fraction. EGFR was found to colocalize with caveolin-1 in transfected cells by confocal laser scanning microscopy and Western blotting results showed that overexpression of caveolin-1 reduced the phosphorylation of EGFR and Erkl/2.

CONCLUSIONOverexpression of caveolin-1 suppresses the growth of HEp2 cells and induces apoptosis and inhibition of EGFR-MAPK signaling pathway may be involved in its mechanism.

Apoptosis ; Blotting, Western ; Carcinoma, Squamous Cell ; genetics ; metabolism ; pathology ; Caveolin 1 ; genetics ; metabolism ; physiology ; Cell Cycle ; Cell Line, Tumor ; Cell Proliferation ; Cell Survival ; Flow Cytometry ; Genetic Vectors ; chemistry ; genetics ; Humans ; Laryngeal Neoplasms ; genetics ; metabolism ; pathology ; Lipids ; chemistry ; Microscopy, Confocal ; Mitogen-Activated Protein Kinase 3 ; metabolism ; Phosphorylation ; Polymerase Chain Reaction ; methods ; Receptor, Epidermal Growth Factor ; metabolism ; Signal Transduction ; physiology ; Transfection ; methods

OBJECTIVETo study whether apoptosis plays a role in controlling the number of olfactory receptor neurons, so as to reveal the specialty and mystery of neurogenesis.

METHODSUsing terminal deoxynucleotidyl transferase-mediated dUTP-fluorescein nick end labeling (TUNEL) and transmission electron microscopy to detect apoptosis in olfactory mucosa of normal adult rats and damaged olfactory mucosa of 16, 32, 48 hours and 3, 7, 30 days after bulbectomy.

RESULTSIn normal olfactory epithelium, a subpopulation of immature neurons, as well as mature neurons, showed internucleosomal DNA-fragmentation. The number of TUNEL-labeled neurons increased dramatically 32 hours after removal of olfactory bulb. Then it declined quickly and remained at low level. Ultrastructural data of olfactory mucosa showed that the feature of apoptotic neurons was chromatin condensation and cell shrinkage. Besides, some dying cells were characterized by the formation of numerous autophagic vacuoles, and few had some of the features of necrosis but without obvious mitochondrial swelling.

CONCLUSIONSApoptosis might play a role in turnover of the olfactory epithelium and regeneration in adult rats. There might be other two types of neural death through different mechanism.

Animals ; Apoptosis ; Male ; Olfactory Bulb ; surgery ; Olfactory Mucosa ; cytology ; pathology ; Olfactory Receptor Neurons ; cytology ; Postoperative Period ; Rats ; Rats, Sprague-Dawley

OBJECTIVESTo analyze the mood factors in prostatitis and evaluate the effects of Molida psychological therapy.

METHODSTwo hundred and thirty-six chronic prostatitis patients were divided according to the course of disease into above 6 months group (n = 31) and below 6 months group(n = 205) as well as into sexual disease group(n = 25) and non-sexual disease group(n = 211). An investigation was made by self-rating method on the basis of the National Institutes of Health Chronic Prostatitis Symptom Index(NIH-CPSI) and Symptom Checklist 90 (SCL-90) and 56 cases were rated again after Molida therapy.

RESULTS1. The scores of SCL-90 in 236 prostatitis patients were significantly higher than normal(P < 0.01). The factor scores of SCL-90 showed one-item positive in 107 patients (45.2%), of whom 27 (25.2%) had depressive disorder and 80(74.77%) had anxiety (23 with significant compulsion). Thirty-eight cases(16.1%) were two items positive. 2. The scores of SCL-90 were significantly higher in the > 6 months group of history and the sexual disease group than in the control group (P < 0.01). 3. The scores of NIH-CPSI showed a positive correlation with those of SCL-90 and both scores in 43/56 cases were significantly decreased after psychological treatment (P < 0.01).

CONCLUSIONSThe mood factor plays an important role in aggravating symptoms in chronic prostatis patients and causes difficulty for management. Molida may significantly improve the mood and symptoms of the chronic prostatitis patient.

Adult ; Affect ; Chronic Disease ; Humans ; Male ; Middle Aged ; Prostatitis ; psychology ; therapy ; Psychotherapy