3.Rapid Detection of Total Bacterial Number Using NADH Fluorescence Method
Jing WANG ; Jing-Xue WANG ; Hong LIN ; Ce-Xia MEI ;
Microbiology 1992;0(05):-
To set up a new method of detecting bacterial number in situ,NADH fluorescence method based on the fluorescent characteristic of NADH was used.When the concentration of NADH ranged from 10 nmol/L to 0.2 mmol/L,its concentration had a good line relationship to the fluorescence intensity(R2= 0.9905).Separating bacterial cells by centrifugation and extracting NADH with hot Tris-HCl buffer,the re-sult of bacterial count detected with NADH standard plot was 1?104 CFU/mL in an hour.In summary,NADH fluorescence method is rapid,sensitive,simple and reliable to detect total bacterial number.There-fore,the method can be widely applied in the field of food sanitation and safety,environment detection and so on.
4.Characters of Respiratory Function in 140 Children with Chronic Cough
hong-xia, LIU ; mei-si, WU ; de-yu, ZHAO
Journal of Applied Clinical Pediatrics 2006;0(16):-
Objective To evaluate the characters of respiratory function in children with chronic cough and explore the correlative association between cough variant asthma(CVA) and etiology of chronic cough.Methods One hundred and forty patients with chronic cough were divided into 2 groups based on peak expiratory flow(PEF) or forced expiratory volume in one second(FEV1).Ninty-three cases were done exercise test and 47 cases were done bronchoditor rest.The parameters included forced vital capacity(FVC),FEV1,PEF,forced expiratory flow at 50% and 75%(FEF50,FEF75).Results The measurement 35 cases with positive bronchoditor rest,and 30 cases with positive exercise test were found.The PEF and FEV1 variation rate were(18.30?10.50)%,(18.78?9.44)% in exercise test groups,and(30.36?27.27)%,(36.13?26.83)% in bronchoditor rest groups,respectively.Conclusions FEV1,PEF may be used as markers for the reversibility of airway obstructe in children with CVA.There is significant correlation between PEF and FEV1.Respiratory function mensurate may reflect the change and degree of inflammation in the airway of children with chronic cough.
5.Early Diagnosis of Alport′s Syndrome in 5 Children and Literature Reviews
li-ping, ZHAO ; bo-ru, MEI ; xiao-hong, GU ; hong-xia, ZHOU
Journal of Applied Clinical Pediatrics 2006;0(17):-
Objective To explore the early diagnosis of Alport′s syndrome(AS).Methods Renal and skin biopsy was carried out in 5 patients who manifested with isolated hematuria and nephritic syndrome(NS).By using indirect immunofluorescence method,the expression of type Ⅳ collagen ? chains was detected on epidermal basement membrane(EBM) and glomerular basement membrane(GBM).Results ?_1 chains on EBM and GBM were expression in all patients,but ?_5 chains on EBM and ?_3,?_5 chains on GBM form 2 female patients were segmental expression.Thus the goal for early diagnosis was achieved.Conclusions ? chains for EBM type Ⅳ collagen and GBM type Ⅳ collegan should be investigated if condition permits for those patients with isolated hematuria,NS(steroid-resistant) and thinned GBM in electron microscopy.It can be useful for diagnosis and differenial diagnosis of AS.
6.Effect of Small Knife Needle on β-enorpin and Enkehalin Contents of Tansverse Process Syndrome of the Third Vertebra.
Nai-gang LIU ; Chang-qing GUO ; Hong-mei SUN ; Xiao-hong LI ; Hai-xia WU ; Hong XU
Chinese Journal of Integrated Traditional and Western Medicine 2016;36(4):476-479
OBJECTIVETo explore the analgesic mechanism of small knife needle for treating transverse process syndrome of the third vertebra (TPSTV) by observing peripheral and central changesof β-endorphin (β-EP) and enkephalin (ENK) contents.
METHODSTotally 30 Japanese white big-ear rabbits of clean grade were divided into 5 groups according to random digit table, i.e., the normal control group, the model group, the small knife needle group, the electroacupunture (EA) group, and the small knife needle plus EA group, 6 in each group. The TPSTV model was established by inserting a piece of gelatin sponge into the left transverse process of 3rd lumbar vertebrae. Rabbits in the small knife needlegroup were intervened by small knife needle. Those in the EA group were intervened by EA at bilateralWeizhong (BL40). Those in the small knife needle plus EA group were intervened by small knife needleand EA at bilateral Weizhong (BL40). Contents of β-EP and ENK in plasma, muscle, spinal cord, and hypothalamus were determined after sample collection at day 28 after modeling.
RESULTSCompared with the normal control group, contents of β-EP and ENK in plasma and muscle increased significantly, and contents of β-EP and ENK in spinal cord and hypothalamus decreased significantly in the model group (P < 0.05, P < 0.01). Contents of β-EP and ENK approximated normal levels in the three treatment groups after respective treatment. Compared with the model group, the content of β-EP in muscle decreased, and contents of β-EP and ENK in hypothalamus increased in the three treatment groups after respective treatment (P < 0.05). There were no significant difference among the three treatment groups (P > 0.05).
CONCLUSIONSSmall knife needle treatment and EA had benign regulation on peripheral and central β-EP and ENK in TPSTV rabbits. Small knife needle treatment showed better effect than that of EA.
Acupuncture Points ; Animals ; Electroacupuncture ; Enkephalins ; metabolism ; Hypothalamus ; metabolism ; Lumbar Vertebrae ; pathology ; Muscle, Skeletal ; metabolism ; Needles ; Rabbits ; Random Allocation ; Spinal Cord ; metabolism ; Spinal Diseases ; therapy ; beta-Endorphin ; metabolism
7.Correlation between p53 gene mutations and p53 protein overexpression in esophageal squamous cell carcinoma.
Guang LI ; Zhao-xia LIU ; Jing HUANG ; Jun-mei WANG ; Quan-hong WANG
Chinese Journal of Pathology 2005;34(12):802-804
Adult
;
Aged
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Carcinoma, Squamous Cell
;
genetics
;
metabolism
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Esophageal Neoplasms
;
genetics
;
metabolism
;
Female
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Gene Expression Regulation, Neoplastic
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Genes, p53
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Humans
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Loss of Heterozygosity
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Male
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Middle Aged
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Point Mutation
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Tumor Suppressor Protein p53
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genetics
;
metabolism
8.Comparison on polysaccharide content and PMP-HPLC fingerprints of polysaccharide in stems and leaves of Dendrobium officinale.
Gui-Fen ZHOU ; Min-Xia PANG ; Su-Hong CHEN ; Gui-Yuan LV ; Mei-Qiu YAN
China Journal of Chinese Materia Medica 2014;39(5):795-802
In order to provide scientific basics for exploitation and sufficient application of Dendrobium officinale leaves resources, the phenol-sulfuric acid method was applied to determine the polysaccharide content. The monosaccharides were derivated by PMP and the derivatives were identified by HPLC-DAD-ESI-MS(n) and the contents of mannose and glucose were determined simultaneously. Similarity evaluation system for chromatographic fingerprint of traditional Chinese medicine (2004A) was employed to generate the mean chromatogram and similarity analysis of the samples was carried out. The results demonstrated that polysaccharide content, monosaccharide compositions and composition ratio had an obvious difference between stems and leaves. The polysaccharide content of stems was higher than that of leaves. Monosaccharide composition in leaf was significantly different from that in stem. The polysaccharide from stems was composed of mannose and glucose, however the polysaccharide of leaves was acid heteropolysaccharide and was mainly composed of five monosaccharides, including mannose, galacturonic acid, glucose, galactose and arabinose. The similarity value of the 14 batches was above 0.9, indicating that similarity of fingerprints among different samples was high. The study can provide evidence for expanding the medicinal parts of D. officinale.
Chromatography, High Pressure Liquid
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Dendrobium
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chemistry
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Mass Spectrometry
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Plant Extracts
;
chemistry
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Plant Leaves
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chemistry
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Plant Stems
;
chemistry
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Polysaccharides
;
chemistry
9.Efficient expression and purification of human beta-defensin-2 in E.coli.
Zhi-xia ZHONG ; Hong RUAN ; Li-mei FAN
Journal of Zhejiang University. Medical sciences 2006;35(6):585-589
OBJECTIVETo demonstrate the possibility of high-level expression of bioactive human beta-defensin-2 (hBD2) in E.coli, and to purify the recombinant hBD2.
METHODSDNA fragment containing mature hBD2 coding region (smhBD2-cDNA) was amplified by PCR, multiple copies of smhBD2-cDNA were linked using Bgl II and BamH I enzymes, pET32-nsmHBD2-cDNA with 1, 2, 4, or 8 copies of smhBD2-cDNA was constructed. The soluble and insoluble hBD2 proteins were separated and analyzed by SDS-PAGE analysis. The soluble protein underwent a separation process containing affinity chromatography, enterokinase digestion and ion exchange chromatography to get the recombinant hBD2 peptide. The bioactivity of recombinant hBD2 was examined by bacteria-inhibition tests in liquid culture.
RESULTThe plasmids pET32-nsmHBD2-cDNA with 1, 2, 4 copies of smhBD2-cDNA were constructed and the expressed soluble protein accounted for 52 %, 48 %, and 31 % respectively. The plasmids with 8 copies expressed mainly insoluble protein with few in soluble form. The growth of E.coli K12D31 was dramatically suppressed with a inhibition rate of 90 %, when the final concentration of recombinant hBD2 reached between 0.4 to 0.5 mug/ml.
CONCLUSIONFusion expression of human beta-defensin-2 with multiple joined genes in E.coli could increase the expression of hBD2.
Anti-Infective Agents ; pharmacology ; Chromatography, Affinity ; Electrophoresis, Polyacrylamide Gel ; Escherichia coli ; drug effects ; genetics ; Gene Expression ; Humans ; Microbial Sensitivity Tests ; Polymerase Chain Reaction ; Recombinant Proteins ; biosynthesis ; isolation & purification ; pharmacology ; beta-Defensins ; biosynthesis ; genetics