1.Optimization of Ethanol Extraction Technology for Xiaoluo Granules by Orthogonal Experiment
Yan WANG ; Weimin CHEN ; Yan HE ; Jing DU ; Yinghua FU
China Pharmacist 2014;(9):1486-1488
Objective:To optimize the ethanol extraction technology for Xiaoluo granules. Methods:The extraction technology for the ethanol dissolvable ingredients in 10 kinds of medicinal materials was optimized by orthogonal experiment. The weighted sum of ba-icalin content and the dry extract rate as the index, 4 influencing factors with 3 levels each were optimized by L9 (34 ) orthogonal table. Results:The optimal ethanol extraction conditions were as follows:the medicinal material was soaked by 10-fold 50% ethanol solution for 45min, and then extracted by heating reflux for 3 times with 1. 5h each time. Conclusion: The method can effectively extract the ethanol dissolvable ingredients in 10 kinds of medicinal materials, which can be used as the ethanol extraction technology for Xiaoluo granules.
2.Evaluation of different tigecycline susceptibility testing methods for Acinetobacter baumannii
Xiaoxing DU ; Haiping WANG ; Ying FU ; Yan CHEN ; Yunsong YU
Chinese Journal of Laboratory Medicine 2013;36(7):598-603
Objective To evaluate different tigecycline susceptibility testing methods for A.baumannii.Methods Thirty carbapenem resistant A.baumannii (CRAB) and 30 carbapenem sensitive A.baumannii (CSAB) isolates were randomly collected from 30 hospitals during January to December in 2010 in China retrospectively.MIC and inhibitory zone diameters for tigecyclinc were determined by the susceptibility testing methods such as broth microdilution (BMD),agar dilution,E test,MIC Test Strip (MTS),Vitek2.Data were analyzed by comparing the results from each method to those produced by the reference BMD method.The effects of two different susceptibility test media (M-H and ISO-Sensitest Agar) on the MIC of tigecycline were also analyzed.Results For CSAB isolates,the MIC50/MIC90 of BMD,agar dilution,E test,MTS and Vitek2 were as follows:0.125/0.25 mg/L,0.125/0.25 mg/L,0.5/1 mg/L,0.125/0.25 mg/L and 0.5/0.5 mg/L.Compared with BMD method,the categorical agreement rates (CA) of each method were ≥90%,and produced no very major errors (VME) by Food and Drug Administration (FDA)/ European Committee on Antimicrobial Susceptibility Testing (EUCAST) breakpoints.For CRAB isolates,the MIC50/MIC90 of BMD,agar dilution,E test,MTS and Vitek2 were as follows:2/4 mg/L,4/4 mg/L,4/4 mg/L,1/2 mg/L and 2/4 mg/L.Compared with BMD method,MTS produced 3.3% (1/30)/6.7% (2/30) VME(FDA/EUCAST breakpoints),and no method CA was ≥90%.The CA between disk diffusion and BMD results were higher by using the criteria of Jones than FDA breakpoints,but only 66.7% (20/30) were observed in CRAB isolates,and produced no VME.The MIC of tigecycline determined using M-H agar were usually higher than those using ISO-sensitest Agar.Conclusions Agar dilution,E test,Vitek 2 and disk diffusion appear not to be a suitable method for routine susceptibility testing of tigecycline for CRAB strains.Tigecycline intermediate or resistant results determined by these methods require confirmation by BMD,and MTS results also need to be interpreted with caution.
3.Influence of PJ34 on the Expression of MMP-9 and Claudin-5 in Rat with Focal Cerebral Ischemia Reperfusion Injury
Penglian WU ; Yan LI ; Xinhui FU ; Lianghe DU ; Dongyu WANG
Journal of China Medical University 2015;44(8):694-698,703
Objective To investigate the effects of PJ34,a poly ADP-ribose polymerase(PARP)inhibitor,on the expression of matrix metallopro-teinases-9( MMP-9 )and Claudin-5 in ischemic cortex of rats with focal cerebral ischemia-reperfusion(I/R)injury. Methods The focal cerebral ischemia-reperfusion model of middle cerebral artery occlusion(MCAO)was established by intraluminal suture . PJ34 was injected intraperitoneal-ly. Blood-brain barrier(BBB)permeability was quantitatively determined by Evans blue assay. Infarct volume changes were observed by 2,3,5-tri-phenyltetrazolium chloridedyeing(TTC)staining. The expression of the MMP-9 and Claudin-5 in rats of cerebral cortex were measured by immuno-histochemistry assay and western blot analysis . Results Compared with sham group,the expression of MMP-9,the contents of EB and infarct vol-ume increased progressively over time after I/R,and reached maximum levels at 48 h(P<0.05);The expression of Claudin-5,the contents of EB and infarct volume reduced significantly over time after I/R,and reached the minimum levels at 48 h in model group(P<0.05). Compared with model group,the expression of MMP-9,the contents of EB and infarct volume was reduced significantly and the expression of Claudin-5,the con-tents of EB and infarct volume was increased at the same time point in PJ34 group(P<0.05). Conclusion PJ34 maintained the blood-brain barri-er permeability by inhibiting the expression of MMP-9,and increasing the expression of Claudin-5,which had neuroprotection on cerebral ischemia-reperfusion injury.
4.Optimizing ethanol extraction technology for no sugar runfei granules by orthogonal experiment design
Lijuan SHEN ; Xingji YANG ; Jing DU ; Yan HE ; Yinghua FU
Chinese Journal of Biochemical Pharmaceutics 2015;(9):158-160,163
Objective To optimize ethonal extraction technology for no sugar runfei granules.Methods Ethanol was used as a solvent to extract the ethanol dissolvable ingredients in 9 kinds of medicinal materials.The formula L9 (34 ) table was used to examine the effects of 4 factors and 3 levels.The weighted sum of baicalin content and the dried extract quantity was used as quantitative index.Results The maximize optimize condition for extraction of ethanol dissolvable ingredients was as follows:9 kinds of medicinal materials, add of 10-fold 70% ethanol solution, soaked for 45 min, and extracted by heating reflux for 3 times,1.5 h each time.Conclusion The method can maximize extraction of ethanol dissolvable effective ingredents in 9 kinds of medicinal materials and can be used as ethonal extraction technologgary of no sugar runfei granules.
5.Influence of Poly (ADP-ribose) Polymerase Inhibitor PJ34 on Blood Brain Barrier in Rats with Cerebral Ischemia-reperfusion Injury
Yan LI ; Penglian WU ; Lianghe DU ; Xinhui FU ; Dongyu WANG
Journal of China Medical University 2015;44(8):709-713
Objective To investigate the influence of poly(ADP-ribose)polymerase inhibitor PJ34 on blood brain barrier(BBB)in rats with cere-bral ischemia-reperfusion injury. Methods Rat model of cerebral ischemia-reperfusion injury was established by the middle cerebral artery occlu-sion. A total of 135 SD rats were randomly divided into 3 groups:sham-operated group(sham group),ischemia-reperfusion group(IR group)and PJ34 group(PJ34 group). 45 animals in each group were then equally divided into subgroups and the rats were sacrificed at 6 h,24 h,48 h after re-perfusion,respectively. BBB permeability was evaluated by detection of extravasated Evans blue(EB). The expression of tumor necrosis factorα(TNF-α)and matrix metalloproteinase 9(MMP-9)activity were measured by immunohistochemistry and western blot at different time points. Re?sults Compared with sham group,the contents of EB and the expressions of TNF-αand MMP-9 in IR group were increased significantly(P<0.05). Compared with IR group,the contents of EB and the expressions of TNF-αand MMP-9 in PJ34 group were markedly decreased at the same time point(P<0.05). Conclusion The present study provided in vivo evidence that PARP inhibitor PJ34 can protect against cerebral ischemia re-perfusion injury,and the mechanism might be related to maintaining the stability of blood-brain barrier by suppressing the expression of TNF-αand MMP-9 in ischemic cortex.
6.Egr-1 promoter regulating effect on granulocyte-macrophage colony-stimulating factor (GM-CSF) gene expression induced by doxorubicin and ionizing radiation
Nan DU ; Xuetao PEI ; Jinming ZHOU ; Junzhong SUN ; Yan FU ; Hui ZHAO
Chinese Journal of Radiological Medicine and Protection 2009;29(3):249-252
Objective To explore the regulating effects of Egr-1 promoter activated by ionizing radiation (IR) and doxorubicin (ADM) on the expression of granulocyte-macrophage colony-stimulating factor (GM-CSF) genes. Methods The human GM-CSF cDNA and enhanced green fluorescent protein (EGFP) cDNA were linked together with IRES(internal ribosome entry site) and then inserted into the expression vector pCIneo under control of the Egr-1 promoter(Egr-EG). The vector was transferred into human bone marrow stromal cell line HFCL by liposome transfection. And the cells were exposure to ADM and IR. The activity of EGFP in HFCL/EG cells were detected by FACS. The effect of N-acetylcysteine on the expression of EGFP following exposure to ADM and IR was examined. The amounts of GM-CSF in HFCL/EG after chemotherapy or radiation were measured with ELISA. The effects of GM-CSF in HFCL/EG cultural supernatants on expansion of CFU-GM derived from cord blood were also studied. RT-PCR analysis for the expression of GM-CSF mRNA in HFCL/EG after exposure to ADM or IR. Results The percentage of EGFP+ HFCL/EG cells exposed to ADM and IR was increased compared with non-treatment group (1.2 % and 15.2 % vs 18.2 %, t = 5.11, P < 0.01). The levels of secreted GM-CSF in HFCL/EG cells exposed to ADM and IR was increased (P < 0.01), but no difference between ADM group and IR group (P 0.05). The expression of EGFP by HFCL/EG treated with ADM and IR was significantly decreased by N-acetylcysteine. The effects of GM-CSF in HFCL/EG cultural supernatants on expansion of CFU-GM in ADM group and IR group were significantly higher than that in HFCL group and non-treatment group. However, The CFU-GM count of IR group was higher than that of ADM group. The expression of GM-CSF mRNA in HFCL/EG cells exposed to ADM and IR was significantly increased(t = 4.37, P < 0.01). Conclusions GM-CSF gene expression regulated by Egr-1 promoter induced by ADM and IR could help the recovery from hematopoietic injury.
7.Evaluation of atopy patch test with dust mite allergens for patients with atopic dermatitis
Yuping FU ; Dongning LI ; Lan WANG ; Jian SUN ; Tiefu YAN ; Lili WANG ; Hongyang DU
Chinese Journal of Dermatology 2016;49(1):40-42
Objective To analyze results of atopy patch test (APT) with dust mite allergens at different concentrations in patients with atopic dermatitis (AD),and to evaluate the diagnostic value of APT.Methods Totally,85 patients with AD were enrolled into this study.All the patients underwent APT with 5 concentrations (3 000,5 000,7 000,10 000 and 12 000 pnu/g) of dust mite allergens,as well as skin prick test (SPT) with dust mite allergens.Dust mite allergens were obtained from two different manufacturers (group 1 and 2).Enzyme-linked immunosorbent assay (ELISA) was performed to detect allergen-specific immunoglobulin E (SIgE) in sera from these patients.The sensitivity,specificity and positive predictive value of APT,SPT and SIgE assay were compared,and the results of APT were compared among different concentrations of allergens and between allergens from different manufacturers.Results When SIgE assay served as the gold standard,the sensitivity,specificity and positive predictive value in the diagnosis of dust mite allergy were 79.41%,76.12%,and 64.29% respectively for APT,73.53%,80.95% and 67.57% respectively for SPT with group 1 dust mite allergens,and 81.53%,77.78% and 65.09% respectively for APT,76.02%,79.85% and 66.07% respectively for SPT with group 2 allergens.When SPT was regarded as the gold standard,the sensitivity,specificity and positive predictive value in the diagnosis of dust mite allergy were 78.38%,77.42%,67.44% respectively for APT,67.57%,84.21%,73.53% respectively for SIgE assay with group 1 dust mite allergens,79.25%,80.63% and 69.55% respectively for APT,61.07%,82.54% and 77.21% respectively for SIgE assay with group 2 allergens.There were no significant differences in the sensitivity,specificity or positive predictive value of APT or SPT between the two groups of allergens.The positive rate of APT was 8.24%,22.35%,29.41%,44.71% and 41.18% respectively with group 1 allergens at 3 000,5 000,7 000,10 000 and 12 000 pnu/g,and 3.53%,23.53%,31.76%,34.12% and 35.29% respectively with group 2 allergens.No significant differences were observed in the positive rate of APT between group 1 and 2 allergens at same concentrations (all P > 0.05),but a significant difference was observed in that between different concentrations of group 1 or 2 allergens (both P< 0.05).The positive rate of APT increased with the increase of allergen concentrations,but stopped rising when the concentrations of group 1 and 2 allergens reached 7 000 pnu/g and 5 000 pnu/g respectively.Conclusions The sensitivity of APT is relatively high for the diagnosis of dust mite allergy.The positive rate of APT increased with the increase in allergen concentrations,but stopped rising when the concentrations reached a certain level.
8.Value of Steel Wire Implantation with Prone Table Stereotactic Digital Mammography in the Detection of Microcalcification of the Breast
Hongbing JIANG ; Jianmin FU ; Fanning ZHANG ; Mu DU ; Yan TENG ; Hong ZHAO ; Manrui CAO ; Enli WANG ; Wenxia ZHANG ; Huan LI
Chinese Journal of Clinical Oncology 2010;37(1):1-4
Objective:To explore the clinical value of stereotactic wire-localized biopsy(SWLB)in digital mammography in the detection of microcalcification of the breast.Methods:A total of 45 patients with nonpalpable breast lesions which were positive for microcalcification by mammography but could not be detected clinically underwent SWLB.Their mammography fndings were analyzed in detail with pathology.Results:Among the 45 cases,13 cases(28.9%)had malignant lesions including ductal carcinoma in situ in 3 cases (20.1%),ductal carcinoma in situ with microinvasion in 4 cases(30.8%),invasive ductal carcinoma in 5 cases (38.5%)and intraductal papillary carcinoma in 1 case(7.7%).Thirty-two cases(71.1%)had benign lesions,2 cases(6.3%)of which were severe atypical hyperplasia.Conclusion:SWLB can accurately guide the surgical excision of nonpalpable breast microcalcification lesions and diagnose microcalcifications exactly,which is helpful for increasing the detection rate of eady-stage breast cancer.
9.Differential proteomic analysis of total protein of ASMC stimulated by wild and mutant IL-13
Xiao-Feng JIANG ; Xiao-Ying GUO ; Hong-Yan LIANG ; Xue-Fei DU ; Yu XIN ; Li XUE ; Song-Bin FU ;
Chinese Journal of Laboratory Medicine 2001;0(04):-
Objective To analyze the differential proteomics of ASMC stimulated by wild IL-13 and mutant IL-13 and to investigate the relations of protein profiles of ASMC to asthma and possible targets for the treatment of bronchial asthma.Methods The total proteins of ASMC stimulated by wild IL-13 and mutant IL-13 were separated by immobilized pH gradient(IPG)-based 2-DE and the differentially expressed protein spots were identified by matrix assisted laser desorption-time of flight mass spectrometry(MALDI-TOF-MS). Results The 2-DE detected approximately(840?21)spots on wild IL-13 samples and(892?17)spots on mutant IL-13 samples(n=3)and(685?19)spots matched.Six significantly differential proteins were subjected to MALDI-TOF-MS analysis and three of them were identified as stathmin 1,Ribosomal protein p~0 and NADH dehydrogenase.Conclusions ASMCs stimulated by wild IL-13 and mutant IL-13 present different proteomic profiles that may shed some light on the mechanism for the asthma causing effect of wild IL-13 and mutant IL-13.
10.Analysis of the difference between the appearance and the bony structure in the polysyndactyly of the fifth toe fused with the fourth toe.
Zuochen DU ; Xiaofei TIAN ; Lin QIU ; Yuexian FU ; Yan LIN ; Xingang YUAN ; Jun XIAO ; Tianwu LI ; Wei CHEN
Chinese Journal of Plastic Surgery 2015;31(2):102-106
OBJECTIVETo investigate difference between the appearance and the bony structure in the polysyndactyly of the fifth toe fused with the fourth toe.
METHODSFrom Jan. 2009 to Jan. 2014, 54 patients (65 feet) with polysyndactyly of the fifth toe fused with the fourth toe were treated. The appearance, X-ray and intraoperative finding were recorded and compared to classify the deformity. Then the extra toe was excised and syndactyly was separated. The malalignment and brachydactyly of the sixth toes were corrected simultaneously.
RESULTSAccording to the bone and joint type, the fifth toes were neoplastic toes without joints in 17 feet, or had poor bony and joint alignment with the sixth toes in 48 feet. So the fifth toes were excised in all the cases. The patients were followed up for 1 month to 4 years. The oblique deformity of sixth toes were corrected completely with improved length.
CONCLUSIONSThe polysyndactyly of the fifth toe fused with the fourth toe should be classified to design the excised toe (usually fifth toe) and correction procedure. The appearance and bony joint recovery are both important.
Humans ; Polydactyly ; pathology ; surgery ; Syndactyly ; pathology ; surgery ; Toe Phalanges ; abnormalities ; surgery ; Toes ; abnormalities ; surgery