【Objective】To explore the optimal conditions in fingerprinting (APHDF).【Methods】The human DNA fingerprints were detected by APHDF.A pair of short primers was used for amplification.The experimental conditions including template,Mg2+,deoxyribonucleotides,and parameters of cycle,were optimized.【Results】The template DNA should be abstracted freshly and the concentration should be ranged from 50～550 mg/L.The best concentration of Mg2+was 5.0 mmol/L.The deoxyribonucleotides concentration was optimal at 0.2 mmol/L.The PCR cycling parameters were as follows :The denaturing temperatures,annealing temperatures and extension temperatures were 94 ℃ and 90 ℃ for 30 s,43 ℃ and 48 ℃ for 40 s or 50 s,and 72 ℃ for 1 min or 80 s,respectively.【Conclusion】The optimal conditions of the experiment are obtained,with good reproducibility and high specificity.Therefore,this method can be widely applied in practice.

ObjectiveTo compare the effect and prognosis of two kinds of internal fixation (improved Giebel blade plate and traditional straddle nail) after high tibial osteotomy (HTO) on osteoarthritis of knee with genu varus deformity. Methods37 knees of 32 cases were treated with straddle nail (25 knees) or Giebel blade plate (12 knees). All the cases were followed up for 6～28 months. ResultsThe clinical bone healing time of osteotomy was 8～12 weeks. There was no significant differences between 2 groups in the increased score in HSS Standard and in the clinical bone healing time. ConclusionBoth internal fixation with improved Giebel blade plate and traditional straddle nail get similarly satisfactory prognosis, while the former shows more advantages to allow early functional exercises.

A simple and rapid method was developed based on high performance liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) to determine sivelestat and its metabolite XW-IMP-A in human plasma. After a simple protein precipitation, the samples and internal standards were analyzed on a C18 column by a gradient elution program. The mobile phase consisted of 30% acetonitrile in methanol and 5 mmol · L(-1) ammonium acetate at a flow rate of 0.7 mL · min(-1). The mass spectrometric data was collected in multiple reaction monitoring mode (MRM) in the negative electrospray ionization. The standard curves were linear in the range of 10.0-15,000 ng · mL(-1) for sivelestat, and 2.50-1000 ng · mL(-1) for XW-IMP-A. The low limits of quantitation were identified at 10.0 and 2.50 ng · mL for sivelestat and XW-IMP-A, respectively. The intra- and inter-day precision were within 11.3% and 13.1% for sivelestat and XW-IMP-A, and accuracy was 0.3% and 0.6% for sivelestat and XW-IMP-A, within the acceptable limits across all concentrations. The method was successfully validated in the pharmacokinetic study of sivelestat in healthy Chinese volunteers.
Chromatography, High Pressure Liquid ; Chromatography, Liquid ; Glycine ; analogs & derivatives ; blood ; Humans ; Inosine Monophosphate ; blood ; Reproducibility of Results ; Sulfonamides ; blood ; Tandem Mass Spectrometry

BACKGROUNDA model of simulated Alzheimer's disease (AD) induced by aggregated amyloid protein (Abeta(1-40)) was built in Wistar rats to observe the behavioral and pathological changes of Abeta(1-40) and the effect of hypodermic insulin injected on the function of study and memory and the expression of Abeta(1-40) from the CA1 area of the hippocampus.

METHODSExperimental groups were as follows: contrast, simulated AD model, contrast of Nacl, and insulin treated. The simulated AD model was built by microinjection of aggregated Abeta(1-40) at the CA1 area of the hippocampus, and was hypodermically injected with 0.9% NaCl (1 ml/kg) and insulin (0.1 U/kg) separately the next day. Two weeks after the modeling, the four groups were tested with water maze about the study and memory function of rats. Three weeks after the injection, the expression of Abeta(1-40) at the CA1 area of the hippocampus was examined by pathological tests (HE, Congo red) and immunohistochemical methods.

RESULTSThe study and memory abilities of rats were ameliorated significantly by the place navigation test and the spatial probe test after the application of insulin. Insulin could decrease the expression of Abeta(1-40) at the CA1 area of the hippocampus to reduce the pathological damage of Abeta(1-40) to the hippocampal area of rats.

CONCLUSIONSThe injection of aggregated Abeta(1-40) to the hippocampal area could simulate the behavioral and pathological features of AD such as the difficulty of study and memory and the damage to neurons. Insulin is effective to improve the function of study and memory and amend the pathological damage of simulated AD model rats. The results give a experimental proof of insulin in the clinical treatment of AD.

Alzheimer Disease ; chemically induced ; drug therapy ; psychology ; Amyloid beta-Peptides ; analysis ; toxicity ; Animals ; Cognition ; drug effects ; Disease Models, Animal ; Hippocampus ; chemistry ; pathology ; Insulin ; pharmacology ; therapeutic use ; Male ; Peptide Fragments ; analysis ; toxicity ; Rats ; Rats, Wistar

OBJECTIVETo study the effect of down-regulation of histone deacetylase 2 (HDAC2) expression on cell proliferation and cell cycle in cervical carcinoma cell lines HeLa.

METHODSHDAC2 siRNA and control siRNA were transfected to HeLa cells. CCK-8 and flow cytometry were used to analyze the changes of cell proliferation and cell cycle, respectively. Western blot was employed to detect the changes of cell proliferation and cell cycle-related proteins.

RESULTSHDAC2 siRNA significantly down-regulated the expression of HDAC2 protein in HeLa cells, resulting in marked inhibition of cell proliferation. In addition, the percentage of cells in G(0)/G(1) phase in HDAC2 siRNA group (63.3% ± 2.0%) was significantly higher than that in untreated group (29.3% ± 1.7%) or control siRNA group (29.4% ± 1.7%), F = 354.181, P = 0.000. Furthermore, Western blot demonstrated that down-regulation of HDAC2 expression decreased the expression of cyclin D1, cyclin E and CDK2 proteins but increased the expression of p21 protein.

CONCLUSIONSDown-regulation of HDAC2 expression mediates proliferation inhibition and cell cycle arrest. It is associated with decrease in cyclin D1, cyclin E and CDK2 protein expression and increase in p21 protein expression.

Cell Cycle ; Cell Proliferation ; Cyclin D1 ; metabolism ; Cyclin E ; metabolism ; Cyclin-Dependent Kinase 2 ; metabolism ; Down-Regulation ; HeLa Cells ; Histone Deacetylase 2 ; genetics ; metabolism ; Humans ; Oncogene Proteins ; metabolism ; Proto-Oncogene Proteins p21(ras) ; metabolism ; RNA, Small Interfering ; genetics ; Transfection

OBJECTIVETo investigate a new technique for functional treatment of chronic facial paralysis.

METHODSBased on anatomy of intramuscular neurovascular structure in the rectus femoris muscle, 7 consecutive patients with facial paralysis were treated by using a technique of microsurgically free-transferring neurovascular rectus femoris muscle segment to the face in one-stage. Follow-ups were 10 to 24 months.

RESULTSAll of the 7 patients showed significantly improvement in the appearance of the oral commissure and oral competence. No complications occurred in the donor site.

CONCLUSIONSThe above mentioned technique may have the advantages of preventing the intramuscular nerve and vessel from the surgical injury during splitting the muscle. It could also maintain the transferred muscular segment in a proper tension in the recipient site.

Facial Paralysis ; surgery ; Follow-Up Studies ; Humans ; Microsurgery ; methods ; Quadriceps Muscle ; blood supply ; innervation ; transplantation ; Reconstructive Surgical Procedures ; Transplant Donor Site ; Treatment Outcome

OBJECTIVEThe objective of this anatomic study was to investigate the intramuscular neurovascular configuration and to evaluate whether the muscle could be split into two functional units in transplantation.

METHODSTen fresh cadavers and ten preserved cadavers were used in the study. A mixture of lead oxide, gelatin and water was injected to the femoral artery of the fresh cadaver. The rectus femoris muscle with its neurovascular pedicles was dissected and radiographed.

RESULTSThree vascular patterns of the rectus femoris muscle were found in the 40 cadaver legs. The muscle received its blood supply through a single vascular pedicle (12.5%), or a dominant pedicle with 1-2 ramified (80%), or two dominant vascular pedicles (7.5%).

CONCLUSIONSThe study provided a detailed description on the intramuscular neurovascular territories of the rectus femoris muscle. Based on the neurovascular supply of the muscle, it is possible to subdivide the muscle into two functional units for segmental muscle transfer.

Cadaver ; Humans ; Quadriceps Muscle ; blood supply ; innervation ; transplantation

OBJECTIVETo investigate the effect of heme oxygenase/carbon monoxide (HO-1/CO) system on lipid deposition at aortic intima and the mechanism involved in hyperlipidemic rabbits.

METHODSTotally 32 rabbits, were divided into four groups. One group as control. Three groups for the following treatments: 1.5% cholesterol ration (Ch group, n = 8); 1.5% cholesterol ration plus HO-1 inducer hemin (Hm group, n = 8); and instead of hemin, the HO-1 inhibitor, zinc protoporphyrin IX (Zn group, n = 8) was given by injection into the abdominal cavity. Experiments were lasted for 12 weeks. Rabbit aortas were then isolated as the samples for histopathologic and ultrastructural examination. The protein expressions of HO-1 and endothelin-1 (ET-1) were investigated by immunohistochemical staining and Western blot analysis.

RESULTSComparing with the Ch group, rabbits of the Hm group showed a remarkably less extent of lipid deposition at the aortic intima [(17.9 ± 3.0)% vs (54.0 ± 4.2)%], and rabbits of the Zn group had a marked extent of lesion development [(61.1 ± 3.5)%]. Lipid deposition, endothelial damage and neo-intimal formation were less severe in rabbits of the Hm group than those in the Zn or Ch group, respectively. Comparing with the control group, rabbits of the Ch group showed a significant decrease of aortic NO production and cNOS activity. However, there were an enhancement of CO production and HO-1 activity (P < 0.01). Compared with Ch group, rabbits of the Hm group showed a remarkable elevation of aortic HO activity and CO production, whereas rabbits of the Zn group showed a marked decrease of both parameters. Compared with the Ch group, rabbits of the Hm group demonstrated a marked reduction of aorta ET-1 expression, whereas Zn group had a significantly higher ET-1 expression.

CONCLUSIONSModulation of HO-1/CO system may improve vascular endothelial function and inhibit smooth muscle cell proliferation in hypercholesterolemic rabbits, likely through a compensatory mechanism and a reduction of ET-1 expression, eventually leading to an inhibition of atherosclerotic plaque development.

Animals ; Aorta ; metabolism ; pathology ; Carbon Monoxide ; metabolism ; Cholesterol ; pharmacology ; Endothelin-1 ; metabolism ; Enzyme Inhibitors ; pharmacology ; Heme Oxygenase-1 ; antagonists & inhibitors ; metabolism ; Hemin ; pharmacology ; Hyperlipidemias ; metabolism ; pathology ; Nitric Oxide ; metabolism ; Nitric Oxide Synthase ; metabolism ; Plaque, Atherosclerotic ; metabolism ; pathology ; prevention & control ; Protoporphyrins ; pharmacology ; Rabbits ; Tunica Intima ; metabolism ; pathology

OBJECTIVETo explore the possibility of repairing chemically induced acute hepatic injuries with allogeneic bone marrow stem cell (BMSC) transplantation.

METHODSA SD rat model of CCl(4)-induced acute hepatic injury was established, which received transplantation of BMSCs (2.0 ml, 1x10(6)/ml) or normal saline injection into the local liver parenchyma, respectively. The rats were sacrificed at 6 h before and 6 h, 1, and 5 weeks after transplantation, and the livers were prepared for microscopic examination.

RESULTSCellular necrosis, bridging necrosis, congestion in the hepatic sinusoid, and inflammatory cell infiltration were seen in the chemically injured livers 6 h after model establishment, and these changes were ameliorated in rats receiving BMSC transplantation.

CONCLUSIONSAllogeneic BMSC transplantation can repair chemically induced acute liver injuries.

Animals ; Bone Marrow Cells ; cytology ; Carbon Tetrachloride ; toxicity ; Chemical and Drug Induced Liver Injury ; etiology ; pathology ; surgery ; Hematopoietic Stem Cell Transplantation ; methods ; Male ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Treatment Outcome

OBJECTIVEPatients with coronary artery disease (CAD, stenosis between 50% - 70% evidenced by coronary angiography) were treated with atorvastatin 40 mg (n = 19) or atorvastatin 10 mg in combination with ezetimibe 10 mg (n = 23). Blood lipid profile and metalloproteinases were monitored up to 3 months.

METHODSCholesterol (TC), triglycerides (TG), low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C), liver function, renal function, creatine kinase, MMP-2, MMP-9, TIMP-1 were measured at baseline and at 1 month and 3 months post therapy.

RESULTS(1) At 3 months, LDL-C was similarly reduced in monotherapy group [(1.94 +/- 0.49) mmol/L, 37.82% reduction compared to baseline] and in combined therapy group [(1.92 +/- 0.54) mmol/L, 38.26% reduction compared to baseline]. (2) AST, ALT, renal function and creatine kinase remained unchanged post various therapy (all P > 0.05). (3) MMP-2, MMP-9 significantly decreased and TIMP-1 significantly increased at 3 months compared to baseline in monotherapy group but these parameters remained unchanged in combined therapy group.

CONCLUSIONBoth therapy regimens were well tolerated and similarly effectively reduced blood lipids and 40 mg atorvastatin monotherapy regimen is superior to atorvastatin 10 mg plus ezetimibe 10 mg regimen in improving metalloproteinases parameters.

Adolescent ; Adult ; Aged ; Anticholesteremic Agents ; administration & dosage ; therapeutic use ; Atorvastatin Calcium ; Azetidines ; administration & dosage ; therapeutic use ; Cholesterol, HDL ; blood ; Cholesterol, LDL ; blood ; Coronary Artery Disease ; drug therapy ; metabolism ; Drug Therapy, Combination ; Ezetimibe ; Female ; Heptanoic Acids ; administration & dosage ; therapeutic use ; Humans ; Hypolipidemic Agents ; administration & dosage ; therapeutic use ; Male ; Metalloproteases ; blood ; Middle Aged ; Pyrroles ; administration & dosage ; therapeutic use ; Tissue Inhibitor of Metalloproteinase-1 ; blood ; Treatment Outcome ; Young Adult