Objective Comparison the coincidence rate in the colloidal gold method and the passive agglu-tination method to detect mycoplasma pneumoniae (MP) infection, discuss the clinical value in rapid diagnosis of MP infection in the two methods. Methods Two-hundred patients with MP infection, including 100 cases in the the children group, and 100 cases in the adult group, were detected in MP-IgM antibody in serum with the colloidal gold method and the passive agglutination method. Results The positive rate of MP-IgM antibody with the passive agglutination method were slightly higher than that of the colloidal gold method in the children group (P > 0.05), While the positive rate of MP-IgM antibody with the passive agglutination method in the adult group were signifi-cantly higher than that of the colloidal gold method (P<0.05). When the antibody titer of MP-IgM antibody were 1:60, ≥1:320 in the children group, the coincidence rate of the positive results with the colloidal gold method and the passive agglutination method were 95.40%, 95.30%;When the antibody titer of MP-IgM antibody were 1:80, 1:160,≥1:320 in the adult group, the coincidence rate of the positive results with the colloidal gold method and the passive agglutination method were 0, 61.90%, 63.80%. Conclusions In the pediatric MP infection, for the high an-tibody titer of MP-IgM antibody, the positive coincidence rate with the colloidal gold method can reach clinical diag-nostic requirements. Clinical physicians according to the age and disease process of patients choose the appropriate method in order to realize the simple, rapid and accurate diagnosis of mycoplasma pneumoniae infection.

Objective To identify and investigate B(A)02 allele in a patient. Methods Serological tests were performed with standard serological methods in a patient with B(A)02 allele. DNA sequences of all seven exons and exon-intron boundaries of ABO gene were analyzed by polymerase chain reaction (PCR), direct DNA sequencing and sequencing after gene cloning. In order to analyze the allele, PyMOL software was used to establish 3D model of Glycosyltransferases B (GTB). Results The serological results showed the characteristics of B(A) phenotype. DNA analysis revealed that ABO gene of the individual was heterozygous of B(A)02/O01 allele. 700C>G mutation was identified in B101 allele, which resulted in the amino acid substitution P234A in GTB. Through the analysis of the 3D structure of GTB, it was speculated that the P234A replacement affected the intermolecular forces of the 234 amino acid and Met-266, thus changed the conformation of the donor-binding pocket of GTB,that made GTB capable of recognizing and tranferring the GalNac to the H antigen, which can lead to the formation of the weak A antigen on membrane of red blood cells. Conclusion The P234A replacement can affect the spatial conformation of the specific recognition region conformed by Met-266 and Ala-268 residues, which leads to the antigenicity change of the ABO blood group.

Objective:To establish a multidrug resistant human salivary gland adenoid cystic carcinoma cell line.Methods:Human salivary gland adenoid cystic carcinoma cell line ACC-2 was treated by 24-hour-exposure to high dose of Bleomycin(BLM)(20 ?g/ml).Drug sensitivity was evaluated by MTT assay.Cell counting was employed to make the growth curve and to calculate the cell doubling time.Flow cytometry(FCM) was used to study the cell cycle distribution and apoptosis.The colony formation ability was also observed.Results:Multidrug resistant cell line of human salivary gland adenoid cystic carcinoma was established and named ACC-2/BLM.After 10 times repeated exposure to BLM,the resistance index(RI) to BLM,5-Fluorouracil(5-FU),Cisplatin(CDDP),Cyclophosphamide(CTX),Vincristine(VCR) were 7.299,1.03,2.15,1.114 and 5.96 respectively.Compared with ACC-2,the proliferation potential of ACC-2/BLM cells was decreased.The ACC-2 apoptosis cells were much more than ACC-2/BLM cells after 9-day-treatment by BLM at 60 ?g/ml.Conclusion:ACC-2/BLM cell line has multidrug resistant characteristics.

A possible association between the polymorphism of glucose transporter 9 (GLUT9) rs1 3137343 and hyperuricemia was investigated in Chinese male population and the calculated genotype frequencies and allelic frequencies by PCR method and direct sequencing were reported.Data showed that there was statistical difference in GLUT9 rs13137343 genotype frequencies between hyperuricemia cases and controls(x2 =7.024,P =0.030).

Objective Idiopathic pulmonary fibrosis ( IPF) is a chronic inflammatory disease with unknown etiology and is lack of effective therapy. The aim of this study is to explore the function of Ca2+ and PI3K/AKT/mTOR pathway in the pathogenesis of IPF, and the impact of 1,25?( OH) 2 D3 on Ca2+ and PI3K/AKT/mTOR pathway in type Ⅱalveolar epithelial cells of rat with IPF. Methods 150 male SD rats were randomly divided into 2 groups: prevention group ( control groupⅠ, model groupⅠ, medication groupⅠ) and treatment group ( control groupⅡ, model groupⅡ, medication groupⅡ) . The tracheal exposure surgery was operated in control groupⅠ/Ⅱ, and then 200μL sterile physiological saline was administered by intraperitoneal injection of each rats 2 days and 14 days after surgery, separately. Bleomycin(BLM)(5 mg/kg) was in?jected into the trachea of model groupⅠ/Ⅱ, and then vitamin D3 solvent(0.1%ethanol and 99.9%glycol propylene, 1μL/g) was ad?ministered by intraperitoneal injection 2 days and 14 days after surger?y, separately. Bleomycin( BLM) ( 5 mg/kg) was injected into the tra?chea of medication groupⅠ/Ⅱ, and then 1,25?( OH) 2 D3( 2μg/kg) was administered by intraperitoneal injection 2 days and 14 days after surgery, separately. IPF model was built by injecting Bleomycin into the trachea of rats, 1,25?(OH)2D3(2μg/kg) was used to prevent and treat IPF by intraperitoneal injection in medication group. The hydroxyproline content of lung tissue in each group was measured, type Ⅱalveolar epithelial cells were separated from lung tissue and labeled with Fluo?3AM, then concentration of Ca2+ was detected by Laser scanning confocal microscope. The mRNA levels of PI3K, AKT and mTOR in the typeⅡalveolar epithelial cells were tested by RT?PCR. Results Compared with control groupⅠ/Ⅱ at each time point, hydroxyproline content of lung tissue, Ca2+ concentration and expression of PI3K, AKT and mTOR in typeⅡalveolar epithelial cells in model groupⅠ/Ⅱand medication groupⅠ/Ⅱwere sig?nificantly raised( P<0.05 or P<0.01) , but these were significantly reduced in medication groupⅠ/Ⅱcompared with model groupⅠ/Ⅱ( P<0.05 or P<0.01) . Correlation analysis showed that there is significant positive correlation between Ca2+ concentration and mRNA expression levels of PI3K, AKT and mTOR in model groupⅠ/Ⅱ(r=0.5988, r=0.6230, r=0.6603,P<0.01)and medication groupⅠ/Ⅱ( r=0.701 2, r=0.632 3,r=0.740 3,P<0.01) . Conclusion The PI3K/AKT/mTOR pathway plays an important role in devel?opment of IPF. 1,25?( OH) 2 D3 is able to reduce Ca2+concentration in typeⅡalveolar epithelial cells and inhibit the PI3K/AKT/mTOR pathway, and then inhibit the development of IPF.

Correlation between the intrahepatic inflammatory pathogenesis and the TCM theory of liver collateral injury by toxins in patients of type 2 diabetes mellitus (T2DM) with insulin resistance (IR) was investigated, to elucidate that removing toxins, dredging collateral and modulating Gan could be one of the effective approaches for inhibiting intrahepatic inflammation mechanism of T2DM with IR.
Chemokine CCL2 ; metabolism ; Diabetes Mellitus, Type 2 ; complications ; metabolism ; physiopathology ; Diagnosis, Differential ; Humans ; Inflammation ; etiology ; metabolism ; physiopathology ; Insulin Resistance ; Liver ; metabolism ; pathology ; physiopathology ; Medicine, Chinese Traditional ; NF-kappa B ; metabolism

The aim of the manuscript was to optimize formulations and preparation technologies of cataplasm of white mustard seed varnish, and to evaluate its anti-asthma effect on rats. The single factor experiments included spreading thickness, types of crosslinking agents, dihydroxyaluminum aminoacetate amount, sodium polyacrylate amount, types of adhesive agents with human sense as the evaluation index. Blank cataplasm matrix was optimized by the orthogonal experiment with the amount of glycerine, citric acid, and sodium carboxymethylcellulose as the major influential factors. Initial adhesive force, peeling strength and human sense were as the evaluation index. The optimized formulation of blank cataplasm were as followings: glycerine-water-ethanol-PEG400-dihydroxyaluminum aminoacetate-citric acid-sodium carboxymethylcellulose-sodium carboxymethylcellulose 2 : 8 : 0.8 : 0.4 : 0.07: 0.15 : 0.1 : 0.5. The active ingredients of white mustard seed, corydalis, and gansui root were extracted by alcohol extraction method. Asiasarum volatile oil was extracted by oil extractor. The optimized drug loading amount was 11% with initial adhesive force, peeling strength and human sense as the evaluation index. Asthma rats model were established by sensitized with ovalbumin and nose-scratching time as the evaluation index. High dose (17%) group of drug-loaded cataplasm had the obvious inhibition effect on nose-scratching time of rats (P = 0.037 < 0.05). In comparison, middle dose (11%), low dose (4%) and positive-control groups had no obvious inhibitive effect on rats. White mustard seed cataplasm supplied a novel choice for anti-asthma therapy. And the overall pharmacodynamics assessment will be carried out on molecular level in near future.
Animals ; Anti-Asthmatic Agents ; administration & dosage ; chemistry ; Asthma ; drug therapy ; Chemistry, Pharmaceutical ; Drugs, Chinese Herbal ; administration & dosage ; chemistry ; Female ; Humans ; Male ; Mustard Plant ; chemistry ; Rats ; Rats, Sprague-Dawley ; Seeds ; chemistry

OBJECTIVETo establish a HPLC-DAD method for the determination of axifolin, naringenin, quercetin and kaempferol in Cudrania tricuspidata and C. cochinchinensis in order to provide a scientific reference for species identification and quality evaluation, by establishing.

METHODThe determination was performed by HPLC-DAD on an Agilent C18 column (4.6 mm x 150 mm, 5 microm) by gradient elution (0-15 min, 35%-50% A; 15-30 min, 50% - 65% A) using methanol (A) and 0.1% phosphoric acid (B) as the mobile phase. The flow rate was 1 mL x min(-1). The detection wavelength was 290 nm for taxifolin and naringenin, 365 nm for quercetin and kaempferol with column temperature at 30 degrees C.

RESULTThe content of axifolin and quercetin in the root of C. tricuspidata were remarkably higher than that in the root of C. cochinchinensis, and the content in stem of C. tricuspidata was also higher than that in the stem of C. cochinchinensis, the content of axifolin and quercetin was variable in different species. The content of naringenin and kaempferol in the root of C. cochinchinensis was visibly higher than that in the root of C. tricuspidata, and the content in the stems of the two herbs was similar, the content of naringenin and kaempferol was visibly variable in different medicinal parts of the herb, but similar between the two herbs.

CONCLUSIONThere's some difference of the content of the four ingredients in different medicinal parts and different herbs, so clinical use should not be confused.

Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Flavanones ; chemistry ; isolation & purification ; Flavones ; chemistry ; isolation & purification ; Kaempferols ; chemistry ; isolation & purification ; Methanol ; Moraceae ; chemistry ; Organ Specificity ; Phosphoric Acids ; Plant Roots ; chemistry ; Plant Stems ; chemistry ; Plants, Medicinal ; Quercetin ; analogs & derivatives ; chemistry ; isolation & purification ; Reproducibility of Results ; Species Specificity

italic>Artemisia argyi (A. argyi) is a Chinese herbal medicine in China. The main active components are volatile oils, flavonoids, and other compounds, which have various pharmacological activities. Methoxylated flavonoids are the main active ingredients in A. argyi. Flavonoid O-methyltransferase (FOMT) is a key enzyme in the O-methylation of flavonoids. In order to further understand the function and characteristics of FOMT proteins, this paper carried out the whole genome mining and identification of FOMT genes in A. argyi and performed phylogenetic, chromosomal localization, gene sequence characterization, subcellular localization prediction, protein structure, gene structure analysis, and expression pattern analysis. The results showed that a total of 83 FOMT genes were identified in the genome of A. argyi. The phylogenetic tree shows that FOMT genes are divided into two subgroups, CCoAOMT (caffeoyl CoA O-methyltransferase) subfamily (32 genes) and COMT (caffeic acid O-methyltransferase) subfamily (51 genes). Gene sequence analysis showed that the number of amino acids encoded by FOMT was 70-734 aa, the molecular weight was 25 296.55-34 241.3 Da, and the isoelectric point was 4.51-9.99. Compared with 32 members of the CCoAOMT subfamily, nearly 1/3 of the 51 members of the COMT subfamily were hydrophobic proteins and 2/3 were hydrophilic proteins. Subcellular localization prediction showed that more than 80% of CCoAOMT subfamily members were located in the cytoplasm, and 96% of COMT subfamily members were located in the chloroplast. COMT subfamily members have more motifs than CCoAOMT subfamily members. The N-terminal motifs of COMT subfamily proteins are relatively variable, while the C-terminal motifs are relatively conserved. Expression pattern analysis showed that CCoAOMT subfamily members were mainly expressed in roots, while COMT members were mainly expressed in leaves. Some FOMTs showed the tissue expression specificity by real-time quantitative PCR analysis, especially in leaves. In this study, we identified and analyzed the FOMT gene family in A. argyi, and provided a theoretical basis for further research on the function of FOMTs and the biosynthesis of methylated flavonoids in A. argyi.

OBJECTIVETo investigate the effect of cardiotrophin-1 (CT-1) on the GATA4 expression and related signaling pathways (JAK-STAT3, ERK1/2 and PI3-K) in rat cardiomyocytes.

METHODSUsing semi-quantitative RT-PCR and EMSA, we measured the dose and time dependent effects of CT-1 on GATA4 mRNA and binding activity in cultured rat cardiomyocytes. Parthenolide (a STAT inhibitor), U-0126 (an ERK inhibitor) and LY-294002 (a PI3-K inhibitor) alone or in combination were added to the culture medium to assess the role of above signaling pathways in CT-1 mediated effects.

RESULTSGATA4 mRNA expression significantly increased at 3 h post 0.1 nmol/L CT-1 exposure, peaked at 6 h and remained high till 24 h post exposure. The GATA4 binding activity began to increase at 10 min and peaked at 60 min and returned to baseline level 180 min. Six hours post CT-1 (0.01 nmol/L, 0.1 nmol/L, 1 nmol/L) exposure, the GATA4 mRNA expression increased in a dose-dependent manner. The GATA4 binding activity peaked with 0.1 nmol/L CT-1 and higher dose did not further increase the binding activity. U-0126 increased the GATA4 mRNA expression and enhanced the GATA4 binding activity and these effects could be partially attenuated with addition of Parthenolide. Parthenolide also prevented the increase of GATA4 mRNA and binding activity induced by CT-1. LY-294002 had no effects GATA4 mRNA and binding activity.

CONCLUSIONCT-1 increases the GATA4 mRNA expression and binding activity in rat cardiomyocytes via STAT3/ERK1/2 pathways and these effects are independent of PI3-K pathway.

Animals ; Cell Line ; Cytokines ; pharmacology ; GATA4 Transcription Factor ; biosynthesis ; genetics ; Myocytes, Cardiac ; metabolism ; RNA, Messenger ; biosynthesis ; Rats ; Rats, Sprague-Dawley ; Reverse Transcriptase Polymerase Chain Reaction ; STAT3 Transcription Factor ; pharmacology ; Signal Transduction