Animals ; Apoptosis ; drug effects ; Caspase 3 ; metabolism ; Hydrocarbons, Brominated ; toxicity ; Isomerism ; Liver ; drug effects ; metabolism ; pathology ; Male ; Rats ; Rats, Sprague-Dawley

Background With the changes of diet and living style,the diabetes has become the major diseases affecting human health.Diabetic cataract is a common complication of diabetes. Objective The present study was to investigate the difference of lens proteomics between diabetic cataract and age related cataract using two dimensional electrophoresis (2-DE) and mass spectrometry in order to postpone happening of diabetic cataract and offer the effective approach to the prevention and therapy of diabetic cataract. Methods The lenses were obtained from 8 diabetic patients and 12 age-related cataract patients during the surgery to extract the protein by lysis and centrifugation.The lens proteins were separated using immobilized pH gradients 2-DE.Image analysis was carried out using PDQuest Advanced-8.0.1 software package.Significant difference of the crystallines was identified by matrixassisted laser adsorption/ionization time of-flight-mass spectrometry (MALDI-TOF-MS) and peptide mass fingerprint combined with protein database. Results The maps of 2-DE showed that lens proteins of diabetic cataract and age related cataract were in the section of pH 5-9 with the relative molecular weight 14000-97000;while relative molecular weight of more abundant crystalline was localized at 20000-31000.About 3 differential protein spots were detected by image analysis software.Two crystallines,αB and βB1 crystallin,were identified using MALDI-TOF-MS.Conclusions Proteomic analysis of lens can be accomplished and the proteins can be well separated,moreover,differential proteins can be analyzed using 2-DE and mass spectrometry between diabetic cataract and age related cataract.These results indicate that αB and βB1 crystallin proteins accelerate the development of diabetic cataract.This technique offers a new avenue for clarity of lens proteins of diabetic cataract other than age related cataract.

Objective To investigate the risk factors and revision strategies for rod fracture in patients with severe kyphoscoliosis following posterior vertebral column resection (PVCR).Methods Between June 2003 and June 2011,7 patients (4 males and 3 females) who developed rod fractures following PVCR in our institution were retrospectively reviewed.The average age was 24.4 years (range,12-39 years) at the time of primary surgeries.The occurrence times and locations of the rod fractures were recorded and the risk factors for the fractures were analyzed.Results The occurrence times of the rod fractures ranged from 6 to 53 months.Five cases of fracture occurred within 2 years after the primary surgeries,while the other 2 cases occurred more than 4 years after the primary surgeries.Six cases of fracture occurred at levels of osteotomies and 1 case occurred at 1 level below the osteotomy.The potential reasons for rod fractures were listed as follows:(1)residual kyphosis (1 case); (2) residual kyphosis combined with unsteady gait (1 case); (3) residual kyphosis combined with single rod fixation (1 case); (4) residual kyphosis combined with malposition of titanium mesh cage (1 case); (5) residual kyphosis combined with anterior column defect; (6) injury (2 cases).Six patients underwent revision surgeries,5 patients received one-stage combined anterior-posterior approach surgeries,while 1 patient received single posterior surgery.They were followed up for 12 to 22 months (average,18months).At final follow-up,all patients obtained satisfying corrections and graft fusion,without internal fixationrelated complications.Conclusion Rod fractures mostly occur at levels of osteotomies within 2 years after PVCR.Residual kyphosis is the main risk factor for the rod fracture.Injury,anterior column defects,unsteady gait,single rod fixation and malposition of titanium mesh cage also increase risks of rod fractures.

ObjectiveTo explore the change of spino-pelvic parameters following pedicle subtraction osteotomy (PSO) for thoracolumbar kyphosis secondary to ankylosing spondylitis(AS).MethodsTwenty-one AS patients with thoracolumbar kyphosis,who underwent PSO at L1 level from July 2006 to October 2010 in our hospital,were retrospectively reviewed.There were 18 males and 3 females with a mean age of 35.6 years (range,21-53 years).The pre- and post-operative thoracic kyphosis(TK),lumbar lordosis (LL),globe kyphosis (GK),angle of the fusion levels (AFL),sagittal imbalance (SVA),pelvic incidence (PI),sacral slope (SS) and pelvic tilting (PT) were measured.ResultsSignificant differences were observed in terms of the improvement of LL,PT,SS,SVA,GK and AFL (P＜ 0.01).The alteration of LL showed significant correlation with the change of PT (r=0.59,P=0.005),SS (r=0.64,P=0.002),SVA (r=0.49,P=0.025),and AFL (r=0.60,P=0.004).The change of PT exhibited cardinal correlation with the change of SS(r=0.94,P=0.000).The improvement of AFL significantly correlated with the improvement of SS(r=0.61,P=0.003),PT (r=0.59,P=0.005).ConclusionThe change of the sagittal spino-pelvic profile following PSO in AS-related thoracolumbar kyphosis is closely related with the improvement of LL.

Objective To investigate the diagnosis and surgical therapy of multiple primary colorectal carcinoma. Methods From 1998 to 2007, 47 patients with synchronous multiple primary colorectal carcinoma and 20 cases with metachronous carcinoma were treated in our hospital. Results In these 67 cases of multiple primary colorectal carcinoma, synchronous carcinoma (SC) accounted for 70% (47 cases) including 37 rectal cancer with a total of 95 larger bowel cancer lesions. There were 6 cases with Dukes A stage, 22 cases with Dukes B stage, 15 cases with Dukes C stage and 4 cases with Dukes D stage. In this whole group there were 20 cases with lymph node metastasis, 21 cases with adenoma and multiple polyps in SC. Three cases received total coloectomy, 10 cases did subtotal coloectomy, 34 cases were treated by radical resection and intestine segment resection. In 20 metachronous carcinoma cases, there were 31 colon cancer(70%) with a total of 44 intestinal cancer lesions. Altogether, there were 17 cases with two tumors, 2 cases with three tumors, one case with four tumors. The duration between the first and the last carcinoma was from 7 months to 19 years, including less than two years in 7 cases, from two to five years in 5 cases, and more than five years in 8 cases. In all 20 MC cases the first (primary) carcinoma received radical resection, while radical resection was performed for the secondary carcinoma in 14 cases and for the third carcinoma in 2 cases. In the SC and the primary carcinoma of MC patients who received radical resection, the 5-year survival rates were 74% and 78% respectively. Conclusion In cases of colonic carcinoma we shouldn't be satisfied with the diagnosis of single colon tumor before a thorough screening of the whole colon was made. In radical resection surgery for SC or MC cases an attempt to preserve enough residual intestinal tract should be made in order to improve the life quality of post-operative patients.

Objective To investigate the expression of guanylyl eyelasec(GC-C)mRNA in pefipheral blood of colorectal cancer patients and discuss its clinical significance. Methods To detect the GC-C mRNA in peripheral blood of colorectal cancer patients by reverse transcriptase PCR and analysis their clinical and pathological indexes. Results The positive rate of GC-C mRNA in 52 colorectal cancer patients was 50%(26/52),while to patients with extra-intestinal cancer,the positive rate was 0(0/8).There were significant difierences of positive rate of GC-C mRNA associated with tumor infiltration(P＜0.05)and tumor embolus in vessel(P＜0.05).While to CEA level in peripheral blood(P＞0.05),but there were no statistical difference.The positive rate of micrometastasis in coloreetal cancer patients might increase using the GC-C mRNA combined with CEA level in peripheral bood.The positive rate of GC-C mRNA in colorectal cancer patients with Dukes A、B、C and D stages were 0(0/3),47.06%(8/17),52.38%(11/21),63.63%(7/11),The expression of GC-C mRNA increased with the clinical stage chang,there was no statistical difference.Conclusions GC-C expresses selectively in peripheral blood of colorectal cancer patients.Itis a valuable new tumor marker in micrometastasis diagnosis and post-operation follow-up of colorectal cancer,and it can also be used as the location determination of metastasis intestinal tumor.

This study is designed to obtain recombinant human acetylcholinesterase (rhAChE) and apply it in screening acetylcholinesterase inhibitors. The rhAChE was overexpressed in HEK293 cells transfected by plasmid of pCMV-AChE with the cationic liposome and rhAChE was found to be secreted into cell culture medium. AChE activity was assayed according to modified Ellman method to obtain kinetic parameters. IC so50 values for donepezil compounds of rhAChE were calculated to determine their activities of inhibition. The results showed that Km value was 151.9 micromol.L-1 donepezil inhibited rhAChE in a mixed competitive-noncompetitive way (Ki= 16.03 nmol.L-1, Ki = 18.36 nmol.L-1) and that most new compounds tested exhibited high activities of inhibition on rhAChE. The study suggests that rhAChE is available to be applied in screening AChE inhibitors in vitro.
Acetylcholinesterase ; genetics ; metabolism ; Cholinesterase Inhibitors ; analysis ; pharmacology ; HEK293 Cells ; Humans ; Indans ; analysis ; pharmacology ; Inhibitory Concentration 50 ; Kinetics ; Piperidines ; analysis ; pharmacology ; Plasmids ; Recombinant Proteins ; genetics ; metabolism ; Transfection

Objective To explore the clinical value of renal dynamic imaging and urinary N-acetyl-?-D-glucosaminidase(NAG),apoptosis DNA fragment(ADF) in evaluating the damage degree of hydronephrotic kidneys(HnK) in children with hydronephrosis.Methods Level of glomerular filtration rate(GFR) was detected in 41 children with congenital hydronephrosis by renal dynamic imaging,and urine NAG,ADF in pelvis in HnK and healthy kidneys (as controls) were detected by enzyme-linked immuno-sorbent assay(ELISA).Patholo-gic changes of HnK in 41 children were graded intoⅠ~Ⅴ according to Elder standard. And GFR,urinary NAG and ADF of HnK were divi-ded into subgroup according to pathologic changes ,at the same time statistical analysis was performed within each groups. And the correlations of pathologic grades with GFR,urinary NAG and ADF of HnK were analyzed.Results 1.Kindneys GFR in healthy kidneys and Hnk were (174.33?20.43)?10-3 L/min,(143.86?17.51)?10-3 L/min respectinely,and there was significant difference between healthy kidneys and Hnk (P0.05).3.There was significant negative correlation between GFR levels of HnK and pathologic grades(r=-0.814 P0.05).Conclusions For hydronephrotic kidneys,urinary NAG can eva-luate impaired nephric tubule whereas renal dynamic imaging may evaluate the damage level of glomeruli;urine ADF may not indicate the damage level of diseased kidneys in children with congenital hydronephrosis.

To study the effects of major components of Maijunan tablets, puerarin (Pue) and rhynchophylline (Rhy) on the transport of hydrochlorothiazide (Hct) Caco-2 cell monolayer model, the transport parameters of Hct, such as apparent permeability coefficient (P(app) (B --> A) and P(app) (A --> B)) and the ratio of P(app) (B --> A) versus P(app) (A --> B), were studied and compared when Hct was used solely and co-used with Pue and/or Rhy. The effects of drug concentrations, conveying times, P-glyprotein (P-gp) inhibitor verapamil and conveying Liq pH values on the transport of Hct in the above conditions were also investigated. The results indicated that the absorption of Hct in Caco-2 cell monolayer model could be a carrier-mediated active transport, along with the excretion action mediated by P-gp. Pue can decrease the excretion action of Hct mediated by P-gp, and Rhy had no significant effect on the transport of Hct. The co-use of Hct, Pue and Rhy enhanced the absorption of Hct. Meanwhile, conveying Liq pH value had significant influence on the transport of Hct. The absorption of Hct at pH 6.0 was higher than that at pH 7.4.
ATP-Binding Cassette, Sub-Family B, Member 1 ; antagonists & inhibitors ; Absorption ; drug effects ; Biological Transport, Active ; drug effects ; Caco-2 Cells ; Drugs, Chinese Herbal ; administration & dosage ; isolation & purification ; pharmacology ; Humans ; Hydrochlorothiazide ; pharmacokinetics ; Hydrogen-Ion Concentration ; Indole Alkaloids ; administration & dosage ; isolation & purification ; pharmacology ; Isoflavones ; administration & dosage ; isolation & purification ; pharmacology ; Plants, Medicinal ; chemistry ; Time Factors ; Vasodilator Agents ; administration & dosage ; isolation & purification ; pharmacology ; Verapamil ; pharmacology