This dissertation is to determine the biopotency of hemostat which processed in different places by establishing a bioassay method of Bletillae Rhizoma based on the thrombin time. Contrast test is the main methodology. Specifically, the reference substance of Bletillae Rhizoma is determined by comparing with the control substance of vitamin K1 using thrombin time, which is calibrated the Bletillae Rhizoma. The hemostatic biopotency is calculated by using the method of "parallel line assay method based on quantitative responses" (3.3) from different processed products. It indicates that there is a strong linear correlation between Bletillae Rhizoma and control drugs (Y = 66.332-23.913X, R2 = 0.995 3). The hemostatic biopotency of Bletillae Rhizoma from different processed products ranged between 821.93-1 187.53 U x g(-1) shown in the paper, and all of them can meet the requirements of the test. The methodology has an appropriate instrument precision (RSD 3.8%), intermediate precision (RSD 4.6%), repeatability (RSD 3.2%) and stability (RSD 3.7%). Therefore, it can be turned out that the methodology which established in the dissertation is good at determinating the hemostatic biopotency of Bletillae Rhizoma and it is reliable, simple and repeatable.
Animals ; Drugs, Chinese Herbal ; pharmacology ; standards ; Hemostatics ; pharmacology ; standards ; Orchidaceae ; chemistry ; Rats ; Rats, Sprague-Dawley ; Rhizome ; chemistry ; Thrombin Time

To establish neonatal rat cardiac fibroblast inflammatory secretion model by using LPS 100 microg x L(-1) combined with ATP 5 mmol x L(-1), in order to study the inhibitory effect of quercetin on the secretion of inflammatory factors TNF-alpha, IL-1beta and IL-6 of cardiac fibroblasts, further investigate the effect of quercetin on the protein expression of p-NF-kappaB p65 (S276) and p-Akt (S473) by western blot, and discuss the inhibitory effect of quercetin on the inflammatory secretion of cardiac fibroblasts. According to the findings, quercetin with the concentrations between 51.74 micromol x L(-1) and 827.81 micromol x L(-1) had no significant effect on the activity of cardiac fibroblasts. Quercetin with the concentrations of 82.78, 41.39, 20.70 micromol x L(-1) could notably inhibit the increase of TNF-alpha and IL-1beta induced by LPS 100 microg x L(-1) for 3 h and then ATP 5 mmol x L(-1) for 36 h (P < 0.01 or P < 0.05). Quercetin with the concentrations of 82.78, 41.39 micromol x L(-1) could notably inhibit the increase of IL-6 induced LPS 100 microg x L(-1) for 3 h and then ATP 5 mmol x L(-1) for 36 h (P < 0.05), without any notable effect of quercetin with the concentration of 20.70 micromol x L(-1). Quercetin with the concentrations of 82.78, 41.39, 20. 70 micromol x L(-1) could notably inhibit the NF-kappaB p65 (S276) activation induced by LPS 100 microg x L(-1) for 3 h and then ATP 5 mmol x L(-1) for 15 min, with the most significant effect in 20.70 micromol x L(-1). Quercetin with the concentrations of 82.78, 41.39, 20.70 micromol x L(-1) could notably inhibit the increase of p-Akt(473) expression induced by LPS 100 microg x L(-1) for 3 h and then ATP 5 mmol x L(-1) for 240 min (P < 0.05). Therefore, this study believes that quercetin could attenuate the secretion of inflammatory factors TNF-alpha, IL-1beta and IL-6 of cardiac fibroblasts by inhibiting the activation of NF-kappaB p65 (S276) and Akt (473).
Animals ; Cells, Cultured ; Drugs, Chinese Herbal ; administration & dosage ; Endomyocardial Fibrosis ; drug therapy ; genetics ; immunology ; Female ; Fibroblasts ; drug effects ; immunology ; Heart ; drug effects ; Humans ; Interleukin-6 ; genetics ; immunology ; Male ; Proto-Oncogene Proteins c-akt ; genetics ; immunology ; Quercetin ; administration & dosage ; Rats ; Rats, Sprague-Dawley ; Tumor Necrosis Factor-alpha ; genetics ; immunology

Objective To investigate the genotype of metallo-?-lactamases (MBL) produced by carbapenem resistant Pseudomonas aeruginosa in pediatric patients.Methods 59 strains of resistance to imipenem or meropenem were collected from December 2003 to November 2005 in Beijing children's hospital.Isolates were further evaluated for MBL production by two screening methods.MBL Etest strips were used to screen the phenotype of MBL production.Molecular screening for blaVIM,blaIMP,blaSPM and blaGIM was carried out using primers targeting the conserved regions of the MBL genes.The PCR fragments obtained with integron primers were sequenced on both strands.The nucleotide sequences were compared with sequences available over the Internet.Results Of the 59 carbapenem resistant Pseudomonas aeruginosa included in this study,29 (49.2%)were MBL positive using Etest methods,and 39 (66.1%) of these tested positive for MBL genes by PCR.35 (89.7%) were positive for blaIMP genes and 4 (10.3%) were positive for blaVIM genes.All isolates were negative for SPM and GIM DNA sequencing revealed that the IMP-1 was detected in 35 IMP-producing isolates,and VIM-2 was detected in 4 VIM-producing isolates.Conclusions This study has demonstrated that MBL-producing strains in pediatric are more common than in adult.IMP-1-producing strains are the main in pediatric,and VIM-2-producing strains concurred.The production of MBL is one of the important reasons of carbapenem resistant Pseudomonas aeruginosa in pediatric.It is very important to monitor the production of MBL.

Background Corneal neovascularization (CNV) is an important cause of visual impairment and graft rejection after allograft corneal transplantation in inflammatory corneal diseases. The mechanisms and therapy relating to CNV are intensely investigated at all times. Objective This study was to evaluate the effect of eicosapentaenoic acid (EPA) on CNV induced by alkali cauterization and its mechanism. Methods The animal models of corneal neovasculation were induced in the right eyes in 72 Sprayue-Dawley rats by putting a piece of 3 mmfilter paper with 1 mol/L NaOH at the center of the cornea for 30 seconds. The rats were then divided randomly into the 0.02 mg EPA treatment group (24 rats) ,0.03 mg EPA treatment group (24 rats) ,model group (24 rats) and normal group (6 rats). EPA of 0.04 ml with doses of 0.02 mg or 0. 03 mg or saline solution of 0. 04 ml was injected subconjunctivally in model rats and immediately after cauterization. The presence of CNV and corneal edema were observed daily by slit lamp biomicroscope. 1,4,7 and 14 days after operation, corneal histopathological examination was performed by hematoxylin and eosin staining. The vascular endothelial cells were stained with CD34 by immunohistochemistry,and the expression of IL-1α,IL-6 mRNA and the nuclear factor-κBp65 ( NF-κBp65 ) proteins was measured by reverse transcription polymerase chain reaction (RT-PCR) and Western blot analysis. The use of animals complied with the Regulations for the Administration of Affair Concerning Experimental Animals by Hebei Province( version 1998 ). Results Under the slit lamp, CNV grew slowly from days 2-4 with obvious corneal edema and defect of epithelium. Larger CNV area and less edema were seen from days 7-10. Maximal vessel growth was observed 14 days after injury with thinner vessels in the model group. Histological examination showed that part of the corneal epithelium was damaged;serious corneal edema, more inflammatory cells and a lot of CNV in the stroma were presented in the model group. However, repairing of the corneal epithelium without CNV ,light corneal edema and less inflammatory cells were found in both the 0. 02 mg EPA and 0. 03 mg EPA treatment groups 7 days after alkali cauterization. The relative area of CNV in the 0. 02 mg EPA treatment group was ( 15.80±6.43 )% and ( 11.06±2. 14)% ,and that in the 0. 03 mg EPA treatment group was (16. 10±7.41 )% and (11.06±2. 51 )%, showing significant reduction in comparison with the model group [ (84. 74±7.77)% and (89.63±7.50) % ] 7 days and 14 days after operation ( P＜0. 05 ). Stronger expression of CD34 in the vascular endothelial cells of the cornea stroma was observed in the model group and an absence of CD34 was observed in the EPA-treated groups on the 7th day. RT-PCR revealed that the expression of IL-1α mRNA and IL-6 mRNA was lower in the EPA treatment groups than the model group ( P＜0. 05 ), and Western blot analysis showed that the expression of NF-κB/p65 in the corneas in the EPA treatment groups was significantly lower than that in the model group on the 4th day after operation (P＜0.05).Conclusion Topical application of EPA suppresses CNV induced by alkali burn possibly by inhibiting the expression of NF-κB,IL-1α and IL-6.

Objective To evaluate the development of the sphincter muscle complex(SMC)and defecation function in pediatric patients with congenital anorectal malformations(ARM).Methods A total of 64 children underwent MRI,among whom 39 were patients with ARM,and the others were patients without ARM undergoing MRI because of other dieases.The dimensions of the SMC in different planes were evaluated with different sequences and coils.The relationship between the SMC development and the defecation function was investigated.Results In control group,the absolute value of SMC width was (3.63?0.22)mm,which had a high correlation with age(r=0.998,P0.05).The SMCs in intermediate ARM patients[muscle index(MI)=0.47?0.05]and low ARM patients(MI=0.49? 0.05)were well developed.The SMCs in a portion of patients with high ARM(MI=0.28?0.06)were poorly developed,when MI≤0.18,anorectal contraction pressurewas significantly lower(t=3.55, P0.18[(0.85?0.20)vs(2.24?1.02)kPa].The length of anal canal with high-pressure[(10.88?3.64)vs(20.26?4.34)mm]was shorter(t=5.18,P0.18,the anorectal angle was less than 90 degrees,and normal continent function was found in 21 of 23 cases(91%).Conclusion MRI can be employed to evaluate the development of SMC in patients with ARM,MI was an objective criteria to evaluate the development of SMC.When MI≤0.18, maldevelopment of SMC will be highly suspected.

Objective To compare the efficacy of transplanting bone marrow mesenchymal stem cell (BMSC) or microenvironmental induced BMSC ( iBMSC) into the ischemic myocardium of rats with myocardial infarction. Methods iBMSC was defined as BMSC co-cultured with myocardial cells for 2 weeks. The stem cells or equal volume PBS were injected into ischemic border zone 1 wk after experimental infarction. Cardiac performance was evaluated at 1, 2, and 4 wk after cell transplantation by echocardiography and analyzed histologically at 4 wk after cell transplantations. Results Compared with PBS group, both BMSC and iBMSC transplantations reduced infarct size. iBMSC enhanced the beneficial effects of BMSC on improving cardiac function (FS: 28.5% ±4.3% in PBS, 29.0% ±2.0% in BMSC and 45. 1% ±3. 1% in iBMSC group at 4 weeks post transplantation, iBMSC group vs. PBS group P <0. 05, iBMSC group vs. BMSC group P <0. 05). Immunofluorescence microscopy results revealed co-localization of SPIO-labeled transplanted cells with cardiac markers for cardiomyocytes, indicating regeneration of damaged myocardium. Conclusion Our data suggest that iBMSC implantation is more effective on improving cardiac function than BMSC implantation in this model. iBMSC might serve as a new promising therapeutic cell source for regenerating ischemic myocardium in patients with post-infarction heart failure.

The clinical evaluation methods of facial paralysis can be divided into functional evaluation scales,neuro-electrophysiologi-cal tests and computer evaluation systems.The commonly used function evaluation scales include House-Brackmann Grading Scale(HB-GS),Burres-Fisch Facial Nerve Scoring System,Nottingham System,Sunnybrook facial grading System(SFGS),Degree of Facial Nerve Paralysis Hierarchical Scale,Facial Disability Index(FDI)and Facial Clinimetric Evaluation(FaCE)Scale,etc.Neuro-electrophysiological tests mainly consist of facial electromyography (EMG), electroneurography (ENoG), blink reflex (BR), and neural excitatory test (NET), etc.The computer evaluation system based on the sensor is mainly divided into the computer evaluation system based on infrared thermal image technology and the computer evaluation system based on biomedicine image recognition.This article briefly summarized the existing methods of facial paralysis evaluation in terms of sensitivity,stability,accuracy,ease of operation and economics.

Congenital heart disease is one of the main types of birth defect.The mammalian heart developmen-tal progress requires precise gene patterning in time and space.In addition to the gene sequence,recent research showed that the regulation of core cardiac gene expression has been proved to be closely related to cardiac transcription factors as well as the modification of genomic architecture of the histone.Methylation of histone might be the key nodes in the regula-tion of cardiac gene expression and chromatin structure.This review focuses on the role of histone H 3 methylation in heart development process,which may lay a foundation for the prediction of epigenetic modification of congenital heart disease.

Objective To investigate the underlying mechanism of the protective effects of resveratrol on oxidant-induced mitochondrial damage in embryonic rat cardiomyocytes.Methods H9c2 cells,a permanent cell line derived from embryonic rat cardiac tissue,and then randomly divided into control group [PBS,cells exposed to H2O2 (600 μmol/L) for 20 min to induce mitochondrial oxidant damage],resveratrol group (0.01,0.1,1,5,10 and 20 μmol/L for 20 min at 20 min before exposing to H2O2),resveratrol plus inhibitor group (1 μmol/L KT5823 for 10 min at 10 min before 5 pmol/L resveratrol treatment) and inhibitor group (1 μmol/L KT5823 for 10 min).Mitochondrial membrane potential (△Ψm) was measured by staining cells with tetramethylrhodamine ethyl ester (TMRE) and the mitochondrial permeability transition pore (mPTP) opening was evaluated by measuring the decrease of TMRE fluorescence intensity.Immunofluorescence assay was used to observe GSK-3β phosphorylation.The phosphorylation of GSK-3β and VASP were determined by Western blot.To detect intracellular NO,cells were loaded with DAF-FM DA (specific fluorescent dye of NO) and imaged with confocal microscopy.Results Compared to the control group,resveratrol (0.01-5 μmol/L) attenuated H2O2-inducedmitochondrial damage reflected by attenuating the H2O2-induced TMRE fluorescence intensity decrease in a dose-dependent manner and the efficacy of 10 and 20 μmol/L resveratrol was significantly lower than that of 5 μmol/L resveratrol.Resveratrol also significantly upregulated the protein expression of VASP and increased GSK-3β Ser9 phosphorylation,which could lead the inactivation of GSK-3β.These effects of resveratrol could be significantly abolished by protein kinase G inhibitor KT5823,while KT5823 alone did not affect CSK-3β and VASP phosphorylation.Confocal microscopy showed that DAF-FM (specific NO indicator) was similar between resveratrol and control group,suggesting that resveratrol did not produce NO.Conclusions Resveratrol could attenuate oxidant-induced mitochondrial damage in embryonic rat cardiomyocytes by inactivating GSK-3β via cGMP/PKG signaling pathway independent of NO-related mechanism.

Objective To study the changes of risk factors of cardiovascular disease in the rural community population after intervention. Methods The Beijing Fangshan cardiovascular prevention program was a community-based comprehensive intervention study which was launched from 1991 and ended in 2000 in five communities including three as intervention communities (IC) and two as control communities (CC) in Fangshan, Beijing suburb. The intervention measures were focused on health education and hypertension control. The changes of risk factors of cardiovascular disease in IC and CC were analyzed using random sample in the year 1991, 1995 and 1999, respectively. The risk factors include systolic and diastolic blood pressure(SBP and DBP), body mass index(BMI), serum total cholesterol(TC), triglyceride(TG),high-density lipoprotein cholesterol(HDL), smoking, and drinking. Results From the year 1991 to 1999, the risk factors of cardiovascular disease such as SBP, DBP, smoking rate and drinking rate were reduced in the population of IC. For male in IC, the decline of SBP, DBP, smoking rate, and drinking rate were 1.6 mmHg, 1.1 mmHg, 14. 5% ( P ＜ 0. 01) and 3.7%, respectively. For female of IC, SBP and DBP declined 4.8 mmHg ( P＜0.01) and 3.2 mmHg ( P ＜0. 01), respectively. SBP, DBP and smoking rate in the population of CC had a little reduction while BMI, TC and TG increased in both IC and CC. During the period of 1991 to 1999, most cardiovascular risk factors in the population of IC had net reduction compared to that of CC. Conclusions Except for BMI and lipids, rural community intervention, as focused on health education and hypertension control, has resulted in the reduction of most risk factors of cardiovascular disease.