[Objective] To investigate associations between the functional polymorphisms of signal transducer and activator of transcription 4 (STAT4) gene and unexplained recurrent spontaneous abortion (URSA) and between STAT4 protein expression and the genotypes of rs 10181656 locus.[Methods] PCR-restriction fragment length polymorphism was used to genotype rs 1 0181656 locus polymorphism in 332 URSA cases and 260 normal controls,in 86 URSA cases and 77 normal controls of which immunohistochemical technique was used to detect STAT4 protein expression.[Results] The frequencies of rs10181656 C/G were 36.45 ％,46.54％ in genotype C/C,46.99％,45.38％ in genotype C/G and 16.57％,8.08％ in genotype G/G between URSA patients and normal controls.They reached statistical difference (P ＜ 0.05).The carriers of rs 10181656 G allele increased the risk of URSA (OR =1.50,P ＜ 0.05).STAT4 protein expression in decidual tissues:①There was statistical difference in the STAT4 protein expression in decidual tissues between cases and controls (P ＜ 0.05).In URSA patients there was statistical difference in the STAT4 protein expression among genotype CC,CG and GG of rs10181656 locus (P ＜ 0.05).So was in normal controls (P ＜ 0.05).In genotype CC there was no difference in the STAT4 protein expression between cases and controls (P ＞ 0.05).Neither was In genotype CG and GG respectively (P all ＞ 0.05).[Conclusion] Functional polymorphisms of the rs10181656 locus could probably associate with the susceptibility of URSA via STAT4 protein expression increased by genotype G/G in maternal decidual tissue.

Objective To explore airway remodeling and air trapping in asthmatic patients with low dose CT scanning and quantitative analysis. Methods 52 stable asthmatic patients in which 29 were severe and 23 were slight,and 20 healthy control cases were underwent low dose dual phase CT scanning. The LA/BSA, WA/BSA, TA/BSA, WA%and Pi10WA were analyzed as airway remolding indexes. The MLD of expiratory, VI-850 (%) of expiratory, MLD E/I, VI-850E-I (%) and VI-850/-950E-I (%) were analyzed as air-trapping indexes. One-Way ANOVA or H Kruskal-Wallis was used to analyze the above indicators. Results Airway remodeling indexes and LA/BSA were (9.6 ± 2.6), (11.0 ± 3.4) and (12.6 ± 3.0)mm2/m2 in severe asthmatics group, non-severe asthmatics group and healthy control group respectively, and there was significant difference between the three groups (F=5.60, P=0.006). WA%of each group was (65.1 ± 2.5)%, (63.3 ± 4.4)%and (62.0 ± 3.0)%, and there was significant difference between the three groups (F=5.53,P=0.006). The Pi10WA was (18.4±2.6), (17.7±3.1) and (16.4±1.4) mm2 respectively with significant difference between the three groups (F=3.59 ,P=0.033). Air-trapping indexes, MLD of expiratory of each group was-(771 ± 59),-(724 ± 43) and-(676 ± 60) HU respectively with significant difference (F=5.60, P=0.006). VI-850(%) of expiratory of each group was 30.79(30.45)%, 13.53(12.09)%and 2.85(6.87)%respectively with significant difference (H=17.20,P<0.001). Conclusions Low dose of CT scan and quantitative analysis can provide an objective and quantitative information for patients with airway disease of asthma, and both WA% and Pi10WA were objective indexes. The severe asthmatic patients were associated with obvious airway remodeling and air trapping compared with non-severe asthmatic patients.

Aged ; Coal Mining ; Humans ; Male ; Middle Aged ; Pneumoconiosis ; complications ; diagnostic imaging ; Pulmonary Heart Disease ; diagnostic imaging ; etiology ; Sensitivity and Specificity ; Tomography, Spiral Computed ; methods

0.05),but a significant difference at weeks 4,8,and 12 between two groups(P

Objective To study the effect of the petroleum ether fraction of ethanol extracts of Polyrhachisvicina Roger with antigout activity on liver microsomal cytochrome P450 in rats.Methods Healthy male SD rats were randomly divided into six groups (3 rats of each group):three groups of the petroleum ether fraction of ethanol extracts of Polyrhachisvi-cina Roger ( test groups, the low dose group, the middle dose group and the high dose group with equivalent to 1, 2, 4 g? kg -1, respectively), two positive control groups and one blank control group.The rats in the low dose, the middle dose and the high dose test groups were adminis-tered daily by gavage at corresponding dose of the petroleum ether frac-tion of ethanol extracts of Polyrhachisvicina Roger for 10 consecutive days.The rats in positive control groups were treated by dexamethasone ( 100 mg? kg-1? d-1 , ip, daily for 4 days ) or phenobarbital ( 80 mg? kg-1? d-1 , ip, daily for 3 days ) .Blank control rats received equivalent volume of sterile normal saline daily by gavage for 10 consecu-tive days.The levels of CYP450 activity, mRNA, protein in rat liver mi-crosome were analyzed by LC/MS/MS or RP-HPLC-FLD, RT-PCR, Western blot, respectively.Results CYP1A2 activity, protein expression and mRNA levels were increased signifi-cantly in a dose-dependent manner with the petroleum ether fraction of ethanol extracts of Polyrhachisvicina Roger at low, middle and high dose, respectively.Conclusion The petroleum ether fraction of ethanol extracts of Polyrhachis-vicina Roger can induce CYP1A2 in rats, suggesting the potential of drug-drug interactions between the petroleum ether fraction of ethanol extracts of Polyrhachisvicina Roger and the inducers, inhibitors, or substrates of CYP1A2.

Objective To explore the expression of P -glycoprotein ( P-gp) in rat liver during acute inflammation ( AI ) and the possible mechanism involved.Methods Twenty rats were randomly divided into two groups:model group (n=10) and control group (n=10).The rats in model group were treated by a single 5 mg? kg -1 intraperitoneal injec-tion of lipopolysaccharide ( LPS).Control rats received equivalent injec-tion of sterile normal saline.The levels of P-gp, ubiquitinated P-gp and 26 S proteasome activity in both groups were analyzed by Western blot, immunoprecipitation and fluorescence detection, respectively. Results Compared to the control groups, ubiquitination levels of liver P-gp and 26S proteasome activity in model groups were significantly in-creased ( P<0.01 ) , accompanied by an decrease of liver P-gp protein expression level. Conclusion The participation of the ubiquitin -proteasome system in down-regulation of liver P-gp expression levels under AI conditions.

To observe a PPAR-alpha agonist effect of N-oleoylethanolamine (OEA) on CB2 (cannabinoid receptor 2), an anti-inflammatory receptor in vascular endothelial cell, healthy HUVECs and TNF-alpha induced HUVECs were used to establish a human vascular endothelial cell inflammatory model. Different doses of OEA (10, 50 and 100 micromol x L(-1)) had been given to HUVECs, cultured at 37 degrees C for 7 h and then collected the total protein and total mRNA. CB2 protein expression was detected by Western blotting and CB2 mRNA expression was assayed by real-time PCR. As the results shown, OEA (10 and 50 micromol x L(-1)) could induce the CB2 protein and mRNA expression, but not 100 micromol x L(-1). To detect if anti-inflammation effect of OEA is partly through CB2, CB2 inhibitor AM630 was used to inhibit HUVEC CB2 expression, then the VCAM-1 expression induced by TNF-alpha was detected, or THP-1 adhere to TNF-alpha induced HUVECs was examined. OEA (50 micromol x L(-1)) could inhibit TNF-alpha induced VCAM-1 expression and THP-1 adhere to HUVECs, these effects could be partly inhibited by a CB2 inhibitor AM630. The anti-inflammation effect of OEA is induced by PPAR-alpha and CB2, suggesting that CB2 signaling could be a target for anti-atherosclerosis, OEA have wide effect in anti-inflammation, it may have better therapeutic potential in anti-inflammation in HUVECs, thus achieving anti-atherosclerosis effect.
Anti-Inflammatory Agents ; pharmacology ; Atherosclerosis ; pathology ; Cell Adhesion ; drug effects ; Cells, Cultured ; Endocannabinoids ; pharmacology ; Endothelial Cells ; cytology ; metabolism ; Ethanolamines ; pharmacology ; Humans ; Indoles ; pharmacology ; Monocytes ; drug effects ; Oleic Acids ; pharmacology ; PPAR alpha ; antagonists & inhibitors ; RNA, Messenger ; metabolism ; Receptor, Cannabinoid, CB2 ; antagonists & inhibitors ; genetics ; metabolism ; Tumor Necrosis Factor-alpha ; pharmacology ; Vascular Cell Adhesion Molecule-1 ; metabolism

Delta6-fatty acid desaturase is a membrane-bound enzyme, which is rate-limiting for the biosynthesis of polyunsaturated fatty acids. A cDNA sequence putatively encoding a delta6-fatty acid desaturase was isolated from Rhizopus arrhizus NK300037 using RT-PCR and RACE methods in our previous work. Sequence and function analysis indicated that this sequence was a novel delta6-fatty acid desaturase gene which had an open reading frame of 1377bp coding 458 amino acids of 52kD. The methylotrophic yeast Pichia pastoris, has been developed into a highly successful system for the production of a variety of heterologous proteins during the past 20 years. In this work, the Rhizopus arrhizus delta6-fatty acid desaturase gene (RAD6) was subcloned into expression vector pPIC3.5K to generate a recombinant plasmid pPICRAD6, which was subsequently transformed into Pichia pastoris strain GS115 for heterologous expression by electroporation method. Total fatty acids were extracted from the induced cells and methylated. The resultant fatty acid methyl esters (FAME) were analyzed by gas chromatography (GC) and gas chromatography-mass spectrometry (GC-MS). Fatty acids analysis showed that the coding product introduced a new double bond at delta6 position of appropriate fatty acid substrates including C16:1, C17:1, C18:1, linoleic acid and alpha-linolenic acid without chain length specificity of fatty acids. Furthermore, modification of sequence flanking AUG codon of this delta6-fatty acid desaturase gene increased the expression of target gene in P. pastoris. All of these results suggest that P. pastoris is an optimal expression system of delta6-fatty acid desaturase gene.
Cloning, Molecular ; Electroporation ; Fungal Proteins ; biosynthesis ; genetics ; Genetic Vectors ; Linoleic Acid ; metabolism ; Linoleoyl-CoA Desaturase ; biosynthesis ; genetics ; Pichia ; genetics ; metabolism ; Rhizopus ; enzymology ; genetics

OBJECTIVETo develop a method for the determination of Chonglou saponin I and Chonglou saponin II in Yifu ointment.

METHODThe chromatographic separation was performed on Hypersil ODS2 C18 column (4.6 mm x 150 mm, 5 microm) and IBBM-612111 ODS C18 guard column. Acetonitrile-0.02% phosphoric (43:57) as mobile phase. The flow rate was 1 mL min(-1) and column temperature was set at 40 degrees C. The UV detection wavelength was set 203 nm.

RESULTChonglou saponin I showed a good linear relationship at a range of 0.1024-3.2 microg, r =0.9998, the average recovery was 97.4%, and RSD was 1.8% (n = 6); Chonglou saponin II showed a good linear relationship at a range of 0.064-2.0 microg, r = 0.9999, the average recovery was 101.4%, and RSD was 1.0% (n =6).

CONCLUSIONThe method is accurate with the good reproducibility and can be used for the quality control of Yifu Ointment.

Chromatography, High Pressure Liquid ; Drug Combinations ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; standards ; Liliaceae ; chemistry ; Ointments ; Plants, Medicinal ; chemistry ; Quality Control ; Reproducibility of Results ; Rosaceae ; chemistry ; Saponins ; analysis

Adult ; Hepatitis B ; prevention & control ; Humans ; Immunity, Active ; Liver Transplantation ; immunology ; methods ; Male ; Postoperative Complications ; prevention & control