1.Study on Optimal Conditions in Arbitrarily Primed PCR Human DNA Fingerprinting
Dayue TONG ; Ping XU ; Yubin GUO ; Fang LI ; Da LIN ; Jinghua OU ; Xinyao WU
Journal of Sun Yat-sen University(Medical Sciences) 2001;22(3):231-234
【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.
2.Comparison of Two Kinds of Internal Fixation after High Tibia Osteotomy for the Treatment of Osteoarthritis of Knee with Genu Varus Deformity
Da-jiang REN ; Fang LI ; Geng CUI ; Quangui WANG ; Zhicheng ZHANG ; Changqing WU ; Juanjuan XU
Chinese Journal of Rehabilitation Theory and Practice 2006;12(5):423-423
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.
3.Simultaneous determination of sivelestat and its metabolite XW-IMP-A in human plasma using HPLC-MS/MS.
Jing WANG ; Xiao-jian DAI ; Yi-fan ZHANG ; Da-fang ZHONG ; Yu-lin WU ; Xiao-yan CHEN
Acta Pharmaceutica Sinica 2015;50(10):1318-1323
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
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Chromatography, Liquid
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Glycine
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analogs & derivatives
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blood
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Humans
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Inosine Monophosphate
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blood
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Reproducibility of Results
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Sulfonamides
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blood
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Tandem Mass Spectrometry
4.Effect of insulin on the cognizing function and expression of hippocampal Abeta1-40 of rat with Alzheimer disease.
Li-hong JIANG ; Yi-na ZHANG ; Xiao-wei WU ; Fang-fang SONG ; Da-yun GUO
Chinese Medical Journal 2008;121(9):827-831
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
5.Effect of down-regulation of histone deacetylase 2 protein expression on cell proliferation and cell cycle in cervical carcinoma.
Fang-fang HUA ; Yong-hua XIA ; Da-peng WU ; Rui-xiang CHEN ; Yu-hong WANG ; Ying PAN ; Jun YANG ; Wu-feng LIANG
Chinese Journal of Pathology 2012;41(7):466-469
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
6.Clinical manifestations and neuroimaging characteristics of children with moyamoya disease.
Jie LI ; Rui LIU ; Zhu-Yi LI ; Da-Fang WU ; Xi-Juan MA ; Jian-Ting MIAO
Chinese Journal of Contemporary Pediatrics 2010;12(8):637-640
OBJECTIVETo study the clinical manifestations and neuroimaging characteristics of pediatric moyamoya disease.
METHODSThe clinical data of 17 children with moyamoya disease were retrospectively studied.
RESULTSThe onset age was between 3 and 14 years. The main clinical manifestations included motor weakness of extremities or hemiplegia, sensory disturbance and headache. Cranial CT or/and MRI examinations predominately showed cerebral infarct. Magnetic resonance angiography (MRA) and digital subtraction angiography (DSA) showed stenosis or occlusion at the terminus of the siphon portions of internal carotid arteries and proximal portions of anterior or middle cerebral arteries, and abnormal vascular networks at the base of brain.
CONCLUSIONSCerebral ischemia is main clinical manifestations in children with moyamoya disease, presenting motor weakness of extremities or hemiplegia, sensory disturbance and headache. DSA is essential to the diagnosis of the disease.
Adolescent ; Angiography, Digital Subtraction ; Child ; Child, Preschool ; Female ; Follow-Up Studies ; Humans ; Magnetic Resonance Imaging ; Male ; Moyamoya Disease ; complications ; diagnosis ; therapy ; Tomography, X-Ray Computed
7.A comparative study of Gd-DTPA and HSA-Gd-DTPA in magnetic resonance lymphography.
Yuan-Kui WU ; Ting-Song FANG ; Da-Chao CHEN ; Guo-Qing LIU ; Yi-Kai XU
Journal of Southern Medical University 2007;27(10):1597-1600
OBJECTIVETo compare the effects of two contrast agents, Gd-DTPA and HSA-Gd-DTPA, in magnetic resonance (MR) lymphography.
METHODSTwelve New-Zealand rabbits were randomized into Gd-DTPA and HSA-Gd-DTPA groups with subcutaneous (interdigital skin fold) injection of the two contrast agents (0.2 ml of 0.5 mmol/L Gd(3+)) for MR lymphography of the popliteal lymph nodes examined in the axial and sagital orientation. T(1)-weighted, T1-weighted fat suppressed, and T(2)-weighted spin-echo (SE) images of the lymph nodes were obtained in plain scans. The post-contrast scanning started at 30 min, 1 h and 3 h after Gd-DTPA administration and at 10 min, 30 min and 60 min after HSA-Gd-DTPA injection to obtain T(1)-weighted images with identical imaging parameters. The signal intensity of popliteal lymph node was measured and the enhancement rate calculated.
RESULTSAfter subcutaneous injection, Gd-DTPA quickly entered blood circulation to result in obvious enhancement of the anterior-tibial vein and the urine and also in heterogeneous enhancement of the popliteal lymph nodes. HSA-Gd-DTPA did not enter the blood, causing obvious homogeneous enhancement of the lymphatic vessels and lymph nodes. HSA-Gd-DTPA resulted in higher enhancement rate than Gd-DTPA, and the enhancement rate in Gd-DTPA group decreased with time as opposed to that of the HSA-Gd-DTPA group.
CONCLUSIONHSA-Gd-DTPA has better performance than Gd-DTPA in MR lymphography after subcutaneous administration.
Animals ; Contrast Media ; administration & dosage ; pharmacokinetics ; Gadolinium DTPA ; administration & dosage ; pharmacokinetics ; Humans ; Lymph Nodes ; diagnostic imaging ; Lymphography ; instrumentation ; methods ; Rabbits ; Random Allocation ; Serum Albumin ; administration & dosage ; pharmacokinetics
8.Battery and power management of portable telemonitoring system.
Wei-jiao LI ; Da-kun LAI ; Xiao-mei WU ; Zu-xiang FANG
Chinese Journal of Medical Instrumentation 2009;33(1):15-19
Biomedical Engineering of Electronic Engineering Deparment, Fudan University Shanghai, 200433 Based on the pulse-coded transmitting and wide-band receiving system, this paper describes a research of phase design a special power management circuit according to the particular demands of a real-time and continuously monitor system is essential. Combined with the requirement of voltage and current in separated module, three aspect must be considered, that is, how to select the functional component, how to accomplish voltage converting and how to carry on battery management. The test at last shows that, using this circuit and the 4400 mAh battery the energy utilization is more than 88%, and the work time is more than 20 hours.
Electric Power Supplies
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Equipment Design
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Monitoring, Ambulatory
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instrumentation
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Monitoring, Physiologic
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instrumentation
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Telemedicine
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instrumentation
9.Effects of different statin regimens on lipid profile and serum metalloproteinases in patients with coronary heart disease.
Wei LIANG ; Hui YANG ; Chun-fang WU ; Qiang YU ; Da-dong ZHANG ; Guo-ping LU
Chinese Journal of Cardiology 2009;37(5):417-421
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
10.Transferring neurovascular rectus femoris muscle segment for treatment of facial paralysis.
Da-ping YANG ; De-hong GUAN ; Xue-wu XU ; Tie-fang GUO ; Xue-feng HAN
Chinese Journal of Plastic Surgery 2003;19(2):101-103
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