With the advance of drug development and research techniques, the drug metabolic processes and mechanism can be more deeply achieved. As the drug metabolism and pharmacokinetics process are mediated by drug metabolizing enzymes and transporters, study of drug metabolizing enzymes and transporters has become an important part for drug development. The traditional immunoassays with low sensitivity and poor specificity can not reflect the accurate expression level of drug metabolizing enzymes and transporters. We now give a brief review on the quantitative study of drug metabolizing enzymes and transporters by mass spectrometry-based proteomic approach.
Enzymes ; chemistry ; Humans ; Inactivation, Metabolic ; Mass Spectrometry ; Membrane Transport Proteins ; chemistry ; Pharmacokinetics ; Proteomics

Objective To observe Meissner's corpuscles in prepuces of different shapes and ages. Methods The Meissner's corpuscles were detected with immunohistochemical stain in 204 prepuce sam- ples of different shapes and ages (3-59 years),and the density of Meissner's corpuscles in every sample was obtained as well.The difference of Meissner's corpuscle densities between phimosis and redundant pre- puce,and correlation between Meissner's corpuscle densities and ages were analyzed with Chi-square test and linear regression,respectively.Results The density of Meissner's corpuscles in redundant prepuce has begun to increase since infancy and reached the peak at the age about 15 years.No significant difference in densities of Meissner's corpuscles between phimosis and redundant prepuce was observed till the age of 20 years,and then there was a trend of disappearance of Meissner's corpuscles in redundant prepuce.A signifi- cantly negative correlation between the densities of Meissner's corpuscles and ages was revealed in redundant prepuce (r=-0.236,P=0.009),whereas an insignificantly positive correlation between the densities of Meissner's corpuscles and ages was shown in phimosis (r=0.193,P=0.084).Conclusions The den- sities of Meissner's corpuscles in redundant prepuce develop synchronically with genital differentiation and accord with the status of sexual function in adult males.The persistent high level of Meissner's corpuscles in adult phimosis might be a mechanism of physiological compensation.

Objective To study the effect of injured podocytes on glomerular maturation and its underlying mechanism in neonatal mice.Methods Single i.p.injection with puromycin aminonucleoside (PA,0.1 mg/g BW) was given to ICR neonatal mice at day 1 after birth (1 dpp). Littermates injected with normal saline (NS) were used as control.Animals were examined for urine protein,blood pressure,kidney weight/body weight (KW/BW),renal histology at 2,4,8,12, 30,60 and 90 dpp (n=6～9 for each group).Immunohistochemistry and quantitative RT-PCR were performed to examine the expression of WT-1,CD31,VEGF,Flk-1,Ang-1,Ang-2,Tie-1 and Tie-2.Results Mice with PA injection had lower kidney weight and body weight at all time points as well as lower KW/BW at 4,8,12 dpp when compared with NS controls.Electron microscopy revealed nearly complete foot process effacement and segmental microvillous transformation as early as 1 day after PA injection.PA-injected kidneys showed fewer capillary loops and decreased maturation index as well as less CD31-positive endothelium in cortical glomeruli at 12 dpp. Glomerular mesangial injury and developing glomerulosclerosis along with proteinuria were noted in PA-injected kidneys starting from 30 dpp.Significantly increased systolic blood pressure was detected at 60 dpp in PA mice.Compared with NS injection,PA injection significantly induced decreased mRNA expression of Flk-1 and Tie-2 as well as increased expression of Ang-1,without obvious changes of VEGF at 2 dpp.Conclusions Podocytes in neonatal kidney of ICR mice are susceptible to PA. Such podocyte injury can alter the expression of VEGF and angiopoietin system in glomeruli,leading to abnormal development of glomerular capillaries,and subsequent proteinuria,hypertension and glomerulosclerosis.

OBJECTIVETo compare therapeutic effects between suspine rotation and flexion manipulation for treating lumbar disc herniation.

METHODSFrom December 2008 to January 2011, 134 patients with lumbar disc herniation were treated, including 74 males and 60 females,ranging in age from 17 to 67 years,33 patients with age over 50 years old. According to diagnosis the ladder of the 9 cases bulging type, 74 hernia type, 51 cases free type. The patients were randomly divided into treatment group ( 69 cases) and control group (65 cases). All the patients were treated with the three-dimensional computer-controlled traction therapeutic apparatus,with continued traction for 20 minutes. After traction, flexion manipulation and rotat-ing manipulation were respectively adopted in treatment group and control group (on alternate days one time, 3 times as a course of treatment). The symptoms and signs (including back pain and discomfort, lower limb pain and numbness, powerless urination and defecation,numbness in perineum, straight-leg raising degree,ability of lower extremity walking,work and live) of patients were observed after treatment.

RESULTSAll patients were followed up for 0.5 to 6 months with an average of 1.1 months. After treatment, the symptoms and signs of patients have significantly improved (P < 0.01), but the symptomes of powerless urination and defecation,numbness in perineum was not improved obviously (P>0.05). Forty-two of 69 patients in treatment group got excellent result, moderate in 20, poor in 7; Twenty of 65 patients in control group got excellent result, moderate in 25,poor in 20.

CONCLUSIONThe whole effect of lumbar backwards rotation manipulation is more satisfactory than flexsion manipulation for treating lumbar intervertebral disc herniation. But spinning force and times should be noticed.

Adolescent ; Adult ; Aged ; Case-Control Studies ; Female ; Humans ; Intervertebral Disc Displacement ; physiopathology ; therapy ; Lumbar Vertebrae ; physiopathology ; Male ; Manipulation, Orthopedic ; methods ; Middle Aged ; Recovery of Function ; Rotation ; Young Adult

Objective:To determine whether ox-LDL (oxdized low-density lipoprotein) is highly deposited on the uremic vessel wall and its influence on the vascular remodeling. Methods: Segments of radial arteries were obtained from 21 uremic subjects during the operation of A-V fistula prior to hemodialysis. Segments of internal thoracic arteries of similar diameter were obtained from patients with benign chest tumors as control.The vascular lesions and ox-LDL, CD68,MCP-1, eNOS,ET-1, PCNA,FN on the vessel wall were determined by means of H-E stain and immunohistochemistry. Results： With H-E stain,atherosclerotic plaques were found in the radial arteries of 4 uremic patients. The middle layer of the arteries in uremic patients were obviously thickened, and the T/D　(thickness of the wall/external diameter) ratio was significantly higher than those in control group(P<0.01). ox-LDL,CD68,MCP-1, ET-1, PCNA,FN on the vessel wall in uremic patients were much higher than those in control group (P<0.01). Moreover, ox-LDL on the vessel wall was positively related to the expression of other above mentioned substances on the vessel wall (P<0.01). Whereas the expression of eNOS on the vessel wall was lower than control group (P<0.01),and was negatively related to ox-LDL on the vessel wall(P<0.01). Conclusion: ox-LDL is an important factor contributing to uremic vascular remodeling by increasing the migration,adhesion and infiltration of monocyte,the proliferation of vascular smooth muscle cell and dysfunction of endothelia.

Objective:To determine whether ox-LDL (oxdized low-density lipoprotein) is highly deposited on the uremic vessel wall and its influence on the vascular remodeling. Methods: Segments of radial arteries were obtained from 21 uremic subjects during the operation of A-V fistula prior to hemodialysis. Segments of internal thoracic arteries of similar diameter were obtained from patients with benign chest tumors as control.The vascular lesions and ox-LDL, CD68,MCP-1, eNOS,ET-1, PCNA,FN on the vessel wall were determined by means of H-E stain and immunohistochemistry. Results： With H-E stain,atherosclerotic plaques were found in the radial arteries of 4 uremic patients. The middle layer of the arteries in uremic patients were obviously thickened, and the T/D　(thickness of the wall/external diameter) ratio was significantly higher than those in control group(P<0.01). ox-LDL,CD68,MCP-1, ET-1, PCNA,FN on the vessel wall in uremic patients were much higher than those in control group (P<0.01). Moreover, ox-LDL on the vessel wall was positively related to the expression of other above mentioned substances on the vessel wall (P<0.01). Whereas the expression of eNOS on the vessel wall was lower than control group (P<0.01),and was negatively related to ox-LDL on the vessel wall(P<0.01). Conclusion: ox-LDL is an important factor contributing to uremic vascular remodeling by increasing the migration,adhesion and infiltration of monocyte,the proliferation of vascular smooth muscle cell and dysfunction of endothelia.

OBJECTIVETo study the expression and significance of nestin (a type of cytoskeletal protein) in normal and diseased human kidneys.

METHODSDiseased kidney tissues were obtained from needle biopsies in 32 patients with glomerulonephritis (including 8 cases of membranous glomerulopathy, 3 cases of focal segmental glomerulosclerosis, 17 cases of IgA nephropathy with proteinuria and 4 cases of IgA nephropathy without proteinuria). Control kidney tissues were obtained from nephrectomy specimens for renal tumors. The expression of nestin in the control kidney tissues was studied using immunoelectronic microscopy and immunohistochemistry. The expression of nestin in the diseased kidney tissues was detected by immunohistochemistry and real-time reverse transcription-polymerase chain reaction.

RESULTSIn normal kidney tissues, nestin was detected at the periphery of glomerular capillary loops. Semi-quantitative morphometric analysis showed that the glomerular nestin expression level in cases of IgA nephropathy without proteinuria did not differ from that in the normal controls. However, the glomerular nestin expression levels in cases of IgA nephropathy with proteinuria, membranous glomerulopathy and focal segmental glomerulosclerosis were significantly lower than those in the normal kidneys and IgA nephropathy without proteinuria. The glomerular nestin expression levels inversely correlated with the 24-hour urine protein results.

CONCLUSIONNestin may play an important role in maintaining the normal function of podocytes in human kidney.

Adult ; Gene Expression Regulation ; Glomerulonephritis, IGA ; immunology ; Humans ; Immunohistochemistry ; Intermediate Filament Proteins ; genetics ; metabolism ; Kidney ; Kidney Diseases ; metabolism ; pathology ; Kidney Glomerulus ; metabolism ; pathology ; Middle Aged ; Nephrectomy ; adverse effects ; Nerve Tissue Proteins ; genetics ; metabolism ; Nestin ; Proteinuria ; metabolism

OBJECTIVETo compare the apical microleakage of Vitapex (calcium hydroxide based paste) with that of AH-plus and zinc oxide eugenol sealer when used with laterally condensed gutta percha obturation technique.

METHODSOne hundred single rooted human anterior teeth were instrumented and randomly divided into three experimental groups (A, B, C) of 30 teeth each and two control groups (D, E) of 5 teeth each. Group A was filled with laterally condensed gutta-percha using Vitapex as sealer. Group B was filled with laterally condensed gutta-percha using AH-plus as sealer. Group C was filled with laterally condensed gutta-percha using zinc oxide eugenol as sealer. Group D was the positive control. Group E was the negative control, which were coated with nail polish to entire root surface. Teeth were then suspended in 2% methylene blue. After this, teeth were demineralized dehydrated and cleared. Linear dye penetration was determined under stereomicroscope with calibrated eye piece.

RESULTSThe mean dye penetration for group A, B, C were respectively (0.57 +/- 0.56) mm, (0.79 +/- 0.96) mm and (1.07 +/- 1.12) mm. Group D demonstrated maximum dye penetration. Group E showed no dye penetration. There was no statistically significant difference between group B and group C (P > 0.05). However, there was statistically significant difference between group A and group B, C (P < 0.01).

CONCLUSIONThis study showed that Vitapex used as endodontic sealer material are better than AH-plus sealer and zinc oxide eugenol sealer.

Calcium Hydroxide ; Dental Leakage ; Dental Pulp Cavity ; Gutta-Percha ; Humans ; Molar ; Root Canal Filling Materials ; Root Canal Obturation ; Silicones ; Zinc Oxide-Eugenol Cement

BACKGROUNDLeft ventricular aneurysm (LVA) is one of the serious complications after acute myocardial infarction. We attempted to evaluate the preliminary efficacy of LVA repair combined with epicardial radiofrequency ablation for ventricular arrhythmia during off-pump coronary artery bypass grafting (OPCAB).

METHODSFrom June 2009 to April 2011, 31 patients with LVA had angina symptoms and ventricular arrhythmia. In all patients, circular and cross-shaped radiofrequency epicardial ablations were performed using unipolar ablation pen along the border between the aneurysm wall and normal cardiac tissue and in the central zone of the aneurysms, followed by a linear placation of ventricular aneurysms on beating heart.

RESULTSAll the patients showed complete recovery. The average number of grafted vessels was 2.7 ± 1.3. Intraoperative examinations revealed that the ventricular arrhythmia was effectively controlled by radiofrequency ablation. All cases had been followed up for one year. Holter monitoring revealed a significant reduction in ventricular arrhythmias (P < 0.05). Echocardiography showed significant increase in left ventricular ejection fraction (P < 0.05) and decrease in left ventricular end-diastolic diameter (P < 0.05).

CONCLUSIONSFor patients with ventricular aneurysm and preoperative malignant arrhythmia, aneurysm repair plus epicardial radiofrequency ablation in OPCAB was found to be an effective and feasible therapeutic technique. However, medium- to long-term therapeutic efficacy of this method remains to be determined by future studies and observations.

Aged ; Arrhythmias, Cardiac ; surgery ; Catheter Ablation ; Coronary Artery Bypass, Off-Pump ; Female ; Heart Aneurysm ; complications ; Heart Ventricles ; surgery ; Humans ; Male ; Middle Aged ; Pericardium ; surgery

AIMTo detect the expression of HSP25 in rat dental follicles both in vivo and vitro, and explore the underlying mechanism of HSP25 on the proliferation and differentiation of rat dental follicle cells (DFCs).

METHODOLOGYImmunohistochemistry was performed to detect the expression of HSP25 in mandibles of postnatal rats on days 1, 3, 5, 7, 9 and 11 in vivo. In vitro, the expression of HSP25 in DFCs was detected by an indirect immunofluorescence assay. Thiazolyl blue tetrazolium bromide (MTT) assay, flow cytometry and alkaline phosphatase (ALP) assay were used to identify the time-course effect mediated by different concentrations of recombinant murine HSP25 of 0, 1, 10, 50 and 100 ng/mL on rat DFCs.

RESULTSExpression of HSP25 was not detected in dental follicles of the rats until day 5 after birth, but became up-regulated in a time-dependent manner till day 11. HSP25 was detected in the cytoplasm of cultured rat DFCs. No significant difference could be observed in the proliferation of DFCs after stimulation with different concentrations of HSP25 on days 1, 2 and 3 (P > 0.05). HSP25 at concentrations of 50 ng/mL and 100 ng/mL up-regulated the ALP activity of DFCs on day 9 (P < 0.05).

CONCLUSIONHSP25-immunoreactivity increased chronologically during the development of dental follicles. The protein had no significant effect on cell proliferation but may play a role in cementoblast/osteoblast differentiation of DFCs.

Alkaline Phosphatase ; analysis ; Ameloblasts ; cytology ; Animals ; Cell Culture Techniques ; Cell Differentiation ; physiology ; Cell Proliferation ; Coloring Agents ; Cytoplasm ; ultrastructure ; Dental Sac ; cytology ; growth & development ; Flow Cytometry ; Fluorescent Antibody Technique, Indirect ; HSP27 Heat-Shock Proteins ; analysis ; physiology ; Odontoblasts ; cytology ; Rats ; Rats, Sprague-Dawley ; Tetrazolium Salts ; Thiazoles ; Tooth Germ ; cytology ; Up-Regulation ; physiology