In-vitro assay methods were established to evaluate transactivation and binding activity of compounds on peroxisome proliferator-activated receptor y (PPARγ). Firstly, plasmids were constructed for transactivation assay of PPARγ response element (PPRE) triggered reporter gene expression, and for cell-based binding activity assay of the chimeric receptor, which was fused with PPARγ ligand binding domain (LBD) and yeast transcriptional activator Gal4. Secondly, by using PPARy competitive binding assay based on time resolved-fluorescence resonance energy transfer (TR-FRET), affinities of compounds and drugs to PPARγ were evaluated. In application of these above methods, the PPARγ activating potency and characteristics of different compounds were evaluated, and a novel benzeneselfonamide derivative, ZLJ01, was found to have comparable binding activity and affinity with the well-known PPARy agonist, but lack of PPRE mediated transactivation activity. In preliminary study on in-vitro hypoglycemic activity, ZLJ1 was found to promote insulin-stimulated glucose uptake by liver cells. Therefore, we believe that combining transactivation and binding activity as well as affinity evaluation, the system could be used to screen non-agonist PPARγ ligand as anovel PPARγ modulator

Objective To assess the clinical safety and feasibility for ultrasound guided paravertebral block anesthesia of percutaneous nephrolithotomy.Methods Between December 2015 to June 2016,180 patients with renal or ureteral calculi were enrolled and evaluated with uhrasonography and CT scan.Of all the 180 patients,108 males and 82 females.Their mean age was 39 years (23-71 years).The clinical characteristics of the patients in each group,such as age,gender,BMI index,ASA status,mean arterial pressure and disease type had no significant differences (P ＞ 0.05).These patients were randomized into general anesthesia group (G group),combined spinal epidural anesthesia group (C group) and paravertebral nerve block anesthesia group (P group).G group:35 males and 25 females.Their mean age was (40.1 ± 11.8) years and BMI was (25.1 ± 3.8) kg/m2;Renal calculi 52 cases,ureteral calculi 8 cases,Average maximum stone diameter was (2.6 ± 0.8)cm.C group:38 males and 22 females.Their mean age was (39.7 ± 12.4) years and BMI was (24.6 ± 4.1) kg/m2;Renal calculi 54 cases,ureteral calculi 6 cases,Average maximum stone diameter was (2.4 ± 0.8) cm.P group:35 males and 25 females.Their mean age was (38.9 ± 12.7) years and BMI was (25.4 ± 4.0) kg/m2;Renal calculi 51 cases,ureteral calculi 9 cases,Average maximum stone diameter was (2.5 ± 0.7) cm.Periprocedural Vital signs,complications,the times of anal discharging gas and postoperative feeding,hospitalized day and expense in these three groups were evaluated.Results Major intraoperative or postoperative complications did not occur in all of the patients.Mean arterial pressure decreased during preoperative changing positions was observed in group G (mean decreased 8.8 mmHg)and group C (mean decreased 1.9 mmHg),with significant difference in intra-group (P ＜ 0.05).Postoperative nausea and vomiting was observed in 8 and 2 patients of group G and group P,respectively (P ＜ 0.05).Postoperative pain was observed in 2 and 7 patients of group C and group P,respectively (P ＞ 0.05).In addition,group P had early post operation feeding time [(6.4 ± 2.4) h],shorter hospitalized day [(4.5 ± 1.1) d] and lower hospitalized expense compared with other groups (P ＜ 0.05).Conclusions Ultrasound guided paravertebral block can provide safe and reliable surgical anesthesia for percutaneous nephrolithotomy.

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.

In-vitro assay methods were established to evaluate transactivation and binding activity of compounds on peroxisome proliferator-activated receptor y (PPARγ). Firstly, plasmids were constructed for transactivation assay of PPARγ response element (PPRE) triggered reporter gene expression, and for cell-based binding activity assay of the chimeric receptor, which was fused with PPARγ ligand binding domain (LBD) and yeast transcriptional activator Gal4. Secondly, by using PPARy competitive binding assay based on time resolved-fluorescence resonance energy transfer (TR-FRET), affinities of compounds and drugs to PPARγ were evaluated. In application of these above methods, the PPARγ activating potency and characteristics of different compounds were evaluated, and a novel benzeneselfonamide derivative, ZLJ01, was found to have comparable binding activity and affinity with the well-known PPARy agonist, but lack of PPRE mediated transactivation activity. In preliminary study on in-vitro hypoglycemic activity, ZLJ1 was found to promote insulin-stimulated glucose uptake by liver cells. Therefore, we believe that combining transactivation and binding activity as well as affinity evaluation, the system could be used to screen non-agonist PPARγ ligand as anovel PPARγ modulator
Genes, Reporter ; Hepatocytes ; Hypoglycemic Agents ; chemistry ; Ligands ; PPAR gamma ; agonists ; chemistry ; Plasmids ; Response Elements ; Sulfonamides ; chemistry ; Transcriptional Activation

The change of the effective components (liquiritin, glycyrrhizic acid, liquiritigenin, isoliquiritigenin) contents of Glycyrrhizae Radix et Rhizoma (GRR) before and after compatibilities in Sini decoction was studied in this paper. Taking single GRR decoction, GRR-Aconiti Lateralis Radix Praeparata (ALRP) decoction, GRR-Zingiberis Rhizoma (ZR) decoction and Sini decoction as test samples, the contents changing of the four effective components of GRR were measured by HPLC. The results showed that the contents of the four effective components of GRR in the single GRR decoction was higher than that in other samples, and the sequence was single GRR decoction > GRR-ZR decoction > GRR-ALRP decoction > Sini decoction. The contents of liquiritin were 11.18, 9.89, 9.67, 9.17 mg · g(-1); the contents of glycyrrhizic acid were 20.76, 15.58, 11.30, 8.52 mg · g(-1); the contents of liquiritigenin were 0.66, 0.57, 0.45, 0.24 mg · g(-1); the contents of isoliquiritigenin were 0.14, 0.07, 0.03, 0.01 mg · g(-1). Therefore, the effective components of GRR decreased obviously after GRR compatibility with ZR providing scientific basis for GRR relieving the strong nature of ZR. The effective components of GRR decreased sharply after GRR compatibility with ALRP providing scientific support for the material foundation research of GRR reducing the toxicity of ALRP. The effective components of GRR decreased further in Sini decoction indicating that the three medicines in Sini decoction were interactional, which reflecting the scientific connotation of the mutual-restraint/mutual-detoxication, mutual-promotion/mutual-assistance compatibilities in Sini decoction.
Chromatography, High Pressure Liquid ; Drug Compounding ; methods ; Drugs, Chinese Herbal ; analysis ; Glycyrrhiza ; chemistry ; Rhizome ; chemistry

OBJECTIVETo study the specific metabonomic profiling of serum from colorectal cancer patients to find out the low molecule metabolites associated intimately with colorectal cancer,and to establish specific metabolic model for the diagnosis of colorectal cancer.

METHODSThe metabonomic profiles of the serum samples from colorectal cancer(CRC) patients(n =31) and healthy adults(n =8) were investigated by gas chromatography-mass spectrometry (GC-MS) technique combined with a commercial mass spectral library for the peak clustering based on metabolites.

RESULTSThirty-four endogenous metabolites including some amino acids, carbohydrates, fatty acids and other intermediate metabolites were identified. By t test statistics with P<0.05, P<0.01 respectively, L-valine, L-threonine, 1-deoxyglucose, glycine and ribitol levels were decreased significantly, but 3-hydroxybutyric acid level was increased significantly in the CRC patient group as compared with healthy adult group. PLS-DA based on these metabolites discriminated two groups for each other. Hierarchical clustering based on above 6 significant differential metabolites revealed that the prediction accuracy of colorectal cancer group was 93.5%.

CONCLUSIONGC-MS technique is an alternative tool for the metabonomic study and would be certainly beneficial to the pathological research, early diagnosis and therapy evaluation of CRC.

Adult ; Aged ; Aged, 80 and over ; Case-Control Studies ; Colorectal Neoplasms ; metabolism ; Female ; Gas Chromatography-Mass Spectrometry ; methods ; Humans ; Metabolomics ; Middle Aged

A proposal is presented to improve the original complicated control of the exposure parameters in the direct radiography system (DR). The interface circuitry of the high-voltage generator is redesigned using RS-232 serial COM port. The data and signals are transmitted between the personal computer and the high-voltage generator in the serial mode. A program package is designed to realize the control of the exposure parameters of the high-voltage generator in DR on the PC machine. Experiment results have shown that the proposed console operates steadily and it is capable of providing a more convenient operation console for the direct radiography system.
Computers ; Equipment Design ; Humans ; Microcomputers ; Radiographic Image Enhancement ; instrumentation ; methods ; Radiographic Image Interpretation, Computer-Assisted ; instrumentation ; methods ; Radiography ; instrumentation ; methods ; Software ; User-Computer Interface

OBJECTIVEWe aim to investigate the sonodynamic effect induced by hydroxyl acetylated curcumin (HAC) on THP-1 macrophages.

METHODSTHP-1 derived macrophages (1 x 10(5) per milliliter) were cultured with HAC at a concentration of 5 µg/mL for 4 h and then exposed to pulse ultrasound treatment (0.5 W/cm2) for 5 min. Six hours later, cell viability analysis was performed with CCK-8 assay, apoptosis and necrosis analysis were detected with Annexin V/PI staining by flow cytometery.

RESULTSThe cell viability of THP-1 macrophage decreased significantly in the group treated with the combination of HAC and ultrasound (P < 0.01), and HAC-SDT induced both apoptosis and necrosis in THP-1 macrophages, the apoptotic rate was higher than the necrotic rate with appropriate conditions, the maximum apoptosis/necrosis ratio was detected in sonodynamic therapy (SDT) group (P < 0.01).

CONCLUSIONhAC-SDT was effective to induce THP-1 macrophages apoptosis.

Apoptosis ; Cell Line ; Cell Survival ; Curcumin ; pharmacology ; Humans ; Macrophages ; cytology ; drug effects ; Necrosis ; Ultrasonics

OBJECTIVETo establish the differential expression profiles for exploring new immune mechanism of hepatitis B virus infection.

METHODSDCs were separated from the bone marrow of healthy mouse and cultured in vitro. Then DCs were stimulated with HBsAg, LPS, TNF-alpha, respectively. Twenty-four hours later, the total RNA of cell was extracted. cDNA microarray was used to compare the differential expression of RNA. The significant differential expression of miRNA after the stimulation of HBsAg was chosen. Subsequently, target genes of the significant differential miRNA were forecasted by using computer software.

RESULTSThere were 30 miRNAs whose significant differential expressions were beyond two times after the stimulation of HBsAg. Among them, 16 miRNAs expressions were increased and 14 miRNAs expressions were decreased. There was significant difference in differential expression of miRNA among the 3 different stimulations. The selected target genes included relevant elements of signal pathway of DC.

CONCLUSIONThe alteration of expression profiles of miRNA was specific after DC was stimulated by HBsAg. The selected target genes further demonstrated that miRNA could play important roles in the immune mechanism of HBV infection.

Animals ; Dendritic Cells ; drug effects ; metabolism ; Gene Expression Profiling ; Hepatitis B Surface Antigens ; pharmacology ; Lipopolysaccharides ; pharmacology ; Male ; Mice ; Mice, Inbred C57BL ; MicroRNAs ; analysis ; Oligonucleotide Array Sequence Analysis ; Tumor Necrosis Factor-alpha ; pharmacology

Objective To evaluate the effect of long-term microwave radiation on the expression of N-methyl-D-aspartate receptor(NMDAR),brain derived neurotrophic factor(BDNF) and related molecules in signal pathways in the hippocampus of rats.Methods Fifty male Wistar rats were exposed to microwave radiation at an average power density of 0,5,10,20 and 30 mW/cm2for 6 min/time,3 times/week,and for 6 weeks,which were sacrificed and the hippocampus was quickly removed at 14 d and 28 d after exposure.The changes in NMDAR (NR1,NR2A,NR2B),postsynaptic density protein(PSD)-95,cortactin,BDNF and tyrosine kinase receptor B (TrkB) in hippocampal neurons of each group were detected by Western blotting and image analysis techniques.Results Compared with the control group,the expressions of related proteins did not change significantly after microwave irradiation of 5 mW/cm2 at each time point.After 20 mW/cm2 microwave radiation,the expression of NR1 was increased at 14 and 28 d (P ＜0.05),the expression of NR2A was increased at 28 d (P ＜ 0.05),but the expression of NR2B was decreased at 14 and 28 d (P ＜ 0.05).At a average power density of 30 mW/cm2,the expressions of NR1,NR2A and PSD-95 and the expression of NR2B were decreased at 14 and 28 d(P ＜0.05),and cortactin,BDNF and TrkB were increased at 14 d after irradiation (P ＜ 0.05).Conclusion The effect of different dosages of long-term microwave radiation on the proteins of NMDAR and its signal pathway related molecules is different.Microwave radiation may affect the NMDAR of postsynaptic information transmission through the BDNF-TrkB signaling pathways,which might play an important role in the impediment of learning and memory function caused by microwave radiation.