1.Study on the Interaction of Gliotoxin with BSA
Jun-Jie CHEN ; Mei YANG ; Lian-Ru ZHANG ; Zhong-Hui ZHENG ; Si-Yang SONG ;
Microbiology 2008;0(08):-
The interaction between Gliotoxin and bovine serum albumin (BSA) was studied by the fluo-rescence, Circular Dichroism (CD) and ultraviolet visible (UV-Vis) techniques. The fluorescent experiment showed that the intrinsic fluorescence of BSA was quenched by the binding of gliotoxin in a static quenching procedure, with an association constant of 7.2?103 L/mol and in hydropobic forces. And the CD spectrum revealed that gliotoxin effected the conformation of BSA by increased the mass of ?-helix.
3.Neuro Electrophysiological Characteristics of Paraneoplastic Peripheral Neuropathy
Dan WANG ; Tao WEI ; Wei-mei DUAN ; Juan HAN ; Lian-mei ZHONG
Journal of Kunming Medical University 2018;39(6):66-70
Objective To study the neuro electrophysiological characteristics of paraneoplastic peripheral neuropathy (PPN) . Methods A retrospective study was conducted for 29 PPN patients consecutively referred to Neurology Department of the First Affiliated Hospital of Kunming Medical University between January 2000 and June 2017. The electrophysiological characteristics of motor nerves, sensory nerves of upper and lower limbs were analyzed. Measurement indicators include: (1) The motor conduction velocity and compound muscle action potential amplitude of median, ulnar, tibial, and common peroneal nerves; (2) The sensory conduction velocity and sensory nerve action potential amplitude of median, ulnar, tibial, and superficial peroneal nerves;(3) F waves of median and tibial nerves.Results (1) For patients with PPN, their motor and sensory nerves in upper and lower limbs were damaged. The total anomaly rate of the amplitude was higher than that of the nerve conduction velocity (P<0.05), while the abnormal rate of amplitude of sensory nerve action potential was greater than that of motor nerve compound muscle action potential (P<0.05) . Abnormal motor nerve conduction velocity had a similar incidence to abnormal sensory nerve conduction velocity (P>0.05) . (2) Nerve conduction study showed that abnormality rate of lower extremity is higher than that of upper extremity. (3) Abnormal F waves were observed less frequently than abnormal nerve conduction rates (P<0.05) . Conclusions The electrophysiological properties of PPN were frequently seen in sensorimotor neuropathy. The damage of distal extremities is more serious. The damage in lower extremity is more severe than that in the upper extremity. The axonal damage mainly occurred in sensory nerves. There is no obvious difference in the degree of demyelination between motor and sensory nerves. Evidence can be provided by analyzing the neuro ectrophysiological characteristics for diagnosis of paraneoplastic peripheral neuropathy in early stage.
4.Evaluation of pyrosequencing for the detection of rpoB gene mutation in Mycobacterium tuberculosis.
Rui-juan ZHENG ; Jie WANG ; Lian-hua QIN ; Jun-mei LU ; Zhong-yi HU
Chinese Journal of Preventive Medicine 2008;42(2):77-80
OBJECTIVETo detect the mutations of rpoB gene in Mycobacterium tuberculosis by pyrosequencing and to evaluate the values on detection of rifampin resistance in clinical isolates.
METHODSUsing the new technology of pyrosequencing, the mutations in the rifampin resistance determining region (RRDR) of rpoB gene were analyzed. The results were compared with those obtained from methods of the absolute concentration and the minimum inhibitory concentration (MIC).
RESULTSAmong the 150 Mycobacterium tuberculosis clinical isolates, 84 were susceptible and 66 resistant to RIF. 54 of the 66 resistant isolates were multidrug-resistant (MDR) strains. Ser531Leu and His526Asp or Tyr, including twelve different genotypes and six codons, were the most common mutations. In the drug susceptibility testing, the accordance rates of the pyrosequencing and the absolute concentration method as well as MIC were 92.7% and 97.8% respectively.
CONCLUSIONNot only is the pyrosequencing technology a fast, sensitive and high throughput method in detecting rifampin resistance in Mycobacterium tuberculosis, but also a useful tool in the research of rifampin resistance mechanism.
Bacterial Proteins ; genetics ; DNA-Directed RNA Polymerases ; Drug Resistance, Bacterial ; genetics ; Humans ; Microbial Sensitivity Tests ; Mutation ; Mycobacterium tuberculosis ; drug effects ; genetics ; Phosphoric Acids ; Polymerase Chain Reaction ; Rifampin ; pharmacology
5.Investigation and analysis of occult hepatitis B virus infection among voluntary blood donors in Lishui area.
Yong-Ping JI ; Long-You ZHAO ; Bin ZHOU ; Mei-Lian ZHONG ; Jie ZHUANG ; Quan YUAN
Chinese Journal of Experimental and Clinical Virology 2011;25(6):457-459
OBJECTIVEOccult hepatitis B infection of voluntary blood donors has been plagued in the serum screening. Determined the OBI through the highly sensitive detection methods Nest-PCR among the blood donors, and then learned occult HBV infection and analysed the genotypes of this area.
METHODS10 080 serums of donors were determined respectively by the imported Abbott HBsAg kit and Beijing Wantai anti-HBc and anti-HBs reagents, obtained the gene and detected DNA sequences by the high sensitive Nest-PCR method.
RESULTSAmong 10 080 cases of unpaid blood donors, 108 cases were detected HBsAg positively by Abbott sensitivity kit (positive rate of 1.07%), 767 cases were anti-HBc single - positive (positive rate of 7.67%). 25 patients screened blood donors who tested negative for serum HBsAg and positive for HBV DNA in the 10 080 cases. Occult HBV infection incidence rate was 0.25%. 12 cases were HBV genotype C (48%), 13 cases were genotype B (52%), and no other genotypes. Genotype B has no statistically significant difference to genotype C (P > 0.05). Sequence analysis showed that 5 patients in the HBsAg epitope "a" (aa124 - aa147) have mutation (20%).
CONCLUSIONThe high proportion of occult hepatitis B infected among voluntary blood donors in our country. Also genotype and mutation was differences in different regions.
Adolescent ; Adult ; Blood Donors ; DNA, Viral ; blood ; Female ; Genotype ; Hepatitis B ; epidemiology ; Hepatitis B virus ; classification ; genetics ; Humans ; Male ; Middle Aged ; Mutation ; Sequence Analysis, DNA
6.Effect of dexamethasone on peroxisome proliferator activated receptor-gamma mRNA expression in 3T3-L1 adipocytes with the human recombinant adiponectin.
Qi-mei SHE ; Jing ZHAO ; Xia-lian WANG ; Chang-man ZHOU ; Xian-zhong SHI
Chinese Medical Journal 2007;120(2):155-158
BACKGROUNDThe fat derived protein adiponectin plays an important role in the regulation of glucose metabolism. The aim of this study was to provide the experimental basis for further investigating on adiponectin (ADPN) function. Its eukaryotic recombinant was constructed and expressed in precursor cells of 3T3-L1 adipocytes. The effects of dexamethasone on peroxisome proliferator activated receptor-gamma (PPAR-gamma) mRNA expression in 3T3-L1 cells with human recombinant adiponectin were assessed.
METHODSThe recombinant plasmid pMD18-T-hADPN and eukaryotic expression vector pcDNA3.1(+) were digested by two restrictive endonucleases and adiponectin and linear pcDNA3.1(+) were obtained. Then, they were ligated and translated into JM109. The recombinant pcDNA3.1(+)-hADPN so obtained was identified by digestion by restrictive endonuclease and nucleotide sequencing. The 3T3-L1 precursor cells were transfected using SuperFect Transfection Reagent (Qiagen). Furthermore, 3T3-L1 cells with human recombinant adiponectin incubated with dexamethasone (0.5 mmol/L) for 24 hours, cells were collected and total RNA was extracted. The PPAR-gamma mRNA expression was quantified by semiquantitative reverse transcription-polymerase chain reaction (RT-PCR).
RESULTSAfter eukaryotic recombinant was digested by Hind III and EcoR I, fragments of 800 bp and 5.4 kb were identified by nucleotide sequence scanning and consistent with theoretical values. Electrophoretogram of RT-PCR in 3T3-L1 precursors showed only one band in front of 250 bp, which was consistent with theoretical value 234 bp. In the 3T3-L1 cells, 3T3-L1 cells with plasmid and 3T3-L1 cells human recombinant adiponectin, treatment with dexamethasone (0.5 mmol/L) decreased PPAR-gamma mRNA expression compared to untreated controls (P < 0.01). Effect of dexamethasone on PPAR-gamma mRNA expression in 3T3-L1 cells was reversed by stably transfected human recombinant adiponectin.
CONCLUSIONThe 3T3-L1 cells stably transfected human recombinant adiponectin had increased PPAR-gamma mRNA expression. Dexamethasone suppressed PPAR-gamma mRNA expression in the 3T3-L1 cells. Effect of dexamethasone on PPAR-gamma mRNA expression in 3T3-L1 cells was reversed by stably transfected human recombinant adiponectin.
3T3-L1 Cells ; Adiponectin ; physiology ; Animals ; Dexamethasone ; pharmacology ; Insulin Resistance ; Mice ; PPAR gamma ; genetics ; RNA, Messenger ; analysis ; Reverse Transcriptase Polymerase Chain Reaction
7.Metabonomics Study on Urine 1H-NMR in Chronic Superficial Gastritis Patients with Pi-qi Deficiency Syndrome/Pi-Wei Dampness-heat Syndrome.
Xu-guang SHI ; Zhong-jie ZOU ; Mei-yin WU ; Yuan-gui ZENG ; Zhi-cheng LIAN ; Man-ting HUANG ; Meng-juan GONG
Chinese Journal of Integrated Traditional and Western Medicine 2015;35(12):1427-1432
OBJECTIVETo observe metabolomic changes in urine of chronic superficial gastritis (CSG) patients with Pi-qi deficiency syndrome (PQDS) or Pi-Wei dampness-heat syndrome (PWDHS), thereby providing scientific evidence for syndrome typing of them.
METHODSUrine samples were collected from CSG patients with PQDS/PWDHS and healthy volunteers, 10 in each group. Proton nuclear magnetic resonance spectroscopy (1H-NMR) based metabonomic analysis was performed on urine samples. Contents of related biomarkers were analyzed by principal component analysis (PCA), partial least square discriminant analysis (PLS-DA), and urivariate statistical analysis.
RESULTSPLS-DA analysis showed that metabolites among CSG patients with PQDS/PWDHS and healthy volunteers could be mutually distinguished. Seven differentially identified metabolites were screened from urines of CSG patients with PQDS and healthy volunteers included glutamate, methionine, α-oxoglutarate, dimethylglycine, creatinine, taurine, and glucose. Four differentially identified metabolites were screened from urines of CSG patients with PWDHS and healthy volunteers included 2-hydroxybutyric acid, trimethylamine oxide, taurine, and hippuric acid. Eleven differentially identified metabolites were screened from urines of CSG patients with PQDS and PWDHS included fucose, β-hydroxybutyric acid, alanine, glutamate, methionine, succinic acid, citric acid, creatinine, glucose, hippuric acid, and lactic acid.
CONCLUSIONThe metabolic differences of CSG patients PQDS and PWDHS mainly manifested in glycometabolism, lipid metabolism, and amino acids catabolism, and 1H-NMR based metabonomics may be used in classified study of Chinese medical syndrome typing.
Biomarkers ; urine ; Discriminant Analysis ; Gastritis ; urine ; Hot Temperature ; Humans ; Hydroxybutyrates ; Ketoglutaric Acids ; Least-Squares Analysis ; Medicine, Chinese Traditional ; Metabolome ; physiology ; Metabolomics ; Principal Component Analysis ; Proton Magnetic Resonance Spectroscopy ; Qi ; Syndrome
8.Effect of endogenous TGF-beta1 and TNF-alpha on the As(2)O(3) inducing apoptosis of HL-60 cells.
Yuan-zhong CHEN ; Yong WU ; Mei-juan HUANG ; Lian-huang LU
Chinese Journal of Hematology 2003;24(5):231-234
OBJECTIVETo study the effect of endogenous TGF-beta(1) and TNF-alpha on As(2)O(3) inducing apoptosis of HL-60 cells.
METHODSThe expressions of endogenous TGF-beta(1) and TNF-alpha in apoptotic HL-60 cells induced by As(2)O(3) were assayed by RT-PCR, quantitative RT-PCR, ELISA, DNA fragmentation and TUNEL. The effect of TGF-beta(1) and TNF-alpha antisense phosphorothioate oligodeoxynucleotides (PSODNs) on As(2)O(3) inducing apoptotic HL-60 cells was further studied.
RESULTS(1) Expressions of endogenous TGF-beta(1) and TNF-alpha were significantly up-regulated in As(2)O(3) inducing apoptotic HL-60 cells (from 13,546 +/- 124 and 497,216 +/- 187 before treatment to 23,273 +/- 229 and 674,217 +/- 189 after treatment, respectively), accompanied with down-regulated bcl-2 mRNA expression (from 10,424 +/- 274 before treatment to 3,361 +/- 89 after treatment). (2) TGF-beta(1) and TNF-alpha antisense PSODNs could rescue As(2)O(3) induced apoptosis of HL-60 cells, with a restoration of bcl-2 gene expression.
CONCLUSIONSEndogenous TGF-beta(1) and TNF-alpha played an important role in As(2)O(3) inducing HL-60 cells apoptosis through down-regulation of bcl-2 expression.
Antineoplastic Agents ; pharmacology ; Apoptosis ; physiology ; Arsenicals ; pharmacology ; Down-Regulation ; HL-60 Cells ; Humans ; Oxides ; pharmacology ; Proto-Oncogene Proteins c-bcl-2 ; biosynthesis ; genetics ; RNA, Messenger ; genetics ; Receptors, Tumor Necrosis Factor ; metabolism ; Transforming Growth Factor beta1 ; biosynthesis ; physiology ; Tumor Necrosis Factor-alpha ; physiology
10.Relationship between neonatal cord blood IgE positiveness and their parents allergies as well as birth status
Mei-Ying WANG ; Zhong-An WANG ; Hai-Lian WANG ; Xiao-Jun CHANG ; Li-Qin WANG
Shanghai Journal of Preventive Medicine 2015;(1):24-26
Objective To explore the relationship between neonatal cord blood IgE value and their parents allergies as well as birth status . [ Methods ] The pregnant women in labor were investigated by questionnaire and detected for blood 25-hydroxy vitamin D3 .The neonatal cord blood was collected for detection of IgE in childbirth , and the birth status was traced and recorded .Statistical analysis was done on the correlative factors of umbilical cord blood IgE . [ Results] The facts that mother had history of allergic diseases(OR=1.65), neonatal birth weight was too large (OR=3.36), baby was male (OR=1.90), and born with asphyxia (OR=2.88) were all associated with the positiveness of cord blood IgE (P<0.05). [Conclusion] The allergy history of mother is a risk factor for the rise of cord blood IgE, so measures for eczema prevention should be well taken for her baby .