Toll like receptors (TLRs) are the earliest discovered natural immune pattern recognition receptors (PRRs). The abnormality of TLR signal transduction pathway is the key factor leading to chronic inflammatory, cancer, nervous system disease and cardiovascular diseases. The development of TLR agonists and inhibitors has attracted much attention. Currently known TLR2 agonists, such as lipopeptides or their derivatives, have certain limitations in drug development due to their difficult synthesis, easy hydrolysis, and triggering inflammatory cytokine storms, while inhibitors have been rarely reported. New small molecule TLR2 agonists or inhibitors with higher stability are more likely to be developed as tumor immunotherapy or anti-inflammatory drugs.

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

Objective To induce cord blood derived endothelial progenitor cells(EPCs)into endothelial cells,and investigate the feasibility of these cells as the seed cells of tissue engineering cardiovascular replacement.Methods Mononuclear cells were isolated from fresh cord blood by 6%HES and density gradient centrifugation.Isolated cells were cultured in medium supplemented with vascular endothelial growth factor(VEGF).Attached cells were identied by morphology,immunofluorescence staining and flow cytometry.Results The percentage of mononuclear cells isolated from fresh cord blood was(3.4?2.1)?10~7/mL.The morphology of attached cells changed while being cultured and inducted,from small-sized round cells to spindle-like cells,to a typical cobblestone morphology,and the total number of cells was 10~7.After 7 days of culture,immunofluorescence staining showed that the vWF and VEGFR-2 were expressed.Compared with the original,cell markers CD_133 decreased(3.11%?1.05%) to(0.09%?0.02%),P

By means of in situ hybridization and immunohistochemistry, the protein localization and gene expression of cyclin B1 in spermatogenic cells were characterized during the spermatogenesis of rabbits. The results showed that the cyclin B1 mRNA in rabbit spermatogenic epithelium was expressed dominantly in primary spermatocytes. The expression was observed in round spermatids with a gradual decline in the process of metamorphosis of the spermatids, but not in elongated spermatids and sperms. Cyclin B1 protein was expressed in mitotic spermatogonia and meiotic spermatocytes and was observed predominantly in round and elongated spermatids. These results indicate that the expression of cyclin B1 mRNA and localization of cyclin B1 protein are dependent on the developmental stages of spermatogenic cells.
Animals ; Cyclin B1 ; genetics ; metabolism ; Gene Expression Regulation, Developmental ; Male ; RNA, Messenger ; genetics ; Rabbits ; Spermatids ; metabolism ; Spermatocytes ; cytology ; metabolism ; Spermatogenesis ; genetics ; Spermatogonia ; metabolism

OBJECTIVETo investigate the prognosis of adenoid cystic carcinoma (ACC) in minor salivary glands and its influencing factors.

METHODSClinical data of 52 patients with ACC in minor salivary glands were reviewed. The distribution of stage was as follows: stage I (6%), stage II (21%), stage III (27%) and stage IV (46%). Counting data was analyzed by χ(2) test or Fisher's exact. Survival rates were calculated by Kaplan-Merier method. Statistical significance of differences in the cumulative survival curves was evaluated using the Log-rank test. Multivariate analysis was performed by Cox proportional hazard model.

RESULTSAll patients underwent primary tumor radical resection, 39 patients (75%) received postoperative radiation. The regional recurrence rate was 37% and distant metastasis rate was 21%. The 5-, 10-year cumulative local control rate were 68% and 63% respectively. The 5-, 10-year cumulative distant control rate were 86%, 68% respectively. The 5-, 10-year tumor specific survival rates were 70% and 54% respectively. Multivariate analysis showed that T stage, lymph node metastasis and perineural invasion were relevant to the tumor specific survival of ACC in minor salivary glands.

CONCLUSIONSRecurrence and metastasis were the main cause of treatment failure of ACC in minor salivary glands. T stage, lymph node metastasis and perineural invasion were the independent prognostic factors of ACC in minor salivary glands. Radical surgery and reasonably postoperative radiotherapy were the main treatment strategy.

Adult ; Aged ; Carcinoma, Adenoid Cystic ; pathology ; radiotherapy ; secondary ; surgery ; Cobalt Radioisotopes ; therapeutic use ; Female ; Follow-Up Studies ; Humans ; Lung Neoplasms ; secondary ; Lymph Node Excision ; Lymphatic Metastasis ; Male ; Middle Aged ; Neoplasm Invasiveness ; Neoplasm Recurrence, Local ; Neoplasm Staging ; Particle Accelerators ; Proportional Hazards Models ; Radiotherapy, Adjuvant ; Salivary Gland Neoplasms ; pathology ; radiotherapy ; surgery ; Salivary Glands, Minor ; Survival Rate ; Young Adult

Purpose To analyze the effects of full length and N-terminal fragment of serum response factor (SRF-Full and SRF-N) on TGF-β1-induced differentiation in c-Kit + cardiac stem cells (CSC).Methods Rat SRF-Full and SRF-N (1-254 aa) coding sequences were obtained from cDNA library and cloned into the linearized lentviral vector GV358 (Ubi-MCS3FLAG-SV40-EGFP-IRES-puromycin) to generate the recombinant vectors,and then positive clones were selected and sequenced after transducing the competent cells with recombinant vectors.The recombinant lentvirus were packaged through transfecting the HEK293T cells with SRF-Full,SRF-N overexpressing plasmids and viral packaging plasmids.Neonatal SD rat cKit + CSCs were isolated via magnetic activated cell sorting,and TGF-β1-induced differentiation in SRF-Full and SRF-N overexpression virus-infected CSCs was assessed by quantitative PCR.Results SRF-Full and SRF-N coding sequences were successfully obtained and properly cloned into the linearized GV358.The positive clones were selected and further confirmed by sequencing.With the help of packaging plasmids,the SRFFull and SRF-N overexpressing plasmids-transfected HEK293T cells successfully produced the lentiviral particles with the titer of 2 × 108 TU/mL,and the SRF-Full-Flag and SRF-N-Flag fusion protein were detected by Western blot in virus-infected HEK293T cells.Addition of TGF-β1 significantly induced upregulated mRNAs in cardiomyocyte markers (Nkx2.5,Gata4,cTnI) and smooth muscle cell marker (SM22α) but not the epithelia cell marker (vWF) in CSCs.Overexpression of SRF-Full facilitated TGF-β1-triggered cardiomyocyte differentiation.However,SRF-N exerted anti-differentiation effects in TGF-β1-treated cells.Conclusion The SRF-Full and SRF-N overexpressing recombinant lentiviral vectors are successfully constructed.SRF-Full facilitates while SRF-N suppresses TGF-β1-induced cardiomyocyte differentiation in c-Kit + CSCs.

OBJECTIVETo observe the effects of Tongxinluo Supermicro Powder on the nuclear factor-kappaB (NF-kappaB), inter-cell adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) expression in aorta of rabbits fed with high-lipid diet.

METHODSHealthy male New Zealand rabbits were randomly divided into 4 groups (n = 8 each): control group, model group, atorvastatin group (3 mg x kg(-1) x d(-1) per gavage), and Tongxinluo group (0.31 g x kg(-1) x d(-1) per gavage). At the end of 6 weeks, the expression of NF-kappaB, ICAM-1 and VCAM-1 were observed by immunochemistry methods, Western blotting and reverse transcription polymerase chain reaction (RT-PCR).

RESULTThe nuclear translocation of NF-kappaB in aortic endothelial cells and the gene expressions of NF-kappaB, ICAM-1 and VCAM-1 at protein and mRNA levels of the model group was significantly increased compared that in the control group (all P < 0.05), these effects could be significantly attenuated by atorvastatin and Tongxinluo Supermicro Powder (P < 0.01 vs. model group).

CONCLUSIONSSimilar as atorvastatin, Tongxinluo Supermicro Powder could relieve the process of atherosclerosis by decreasing the nuclear translocation of NF-kappaB and reducing the expression of ICAM-1, VCAM-1 in this model.

Animal Feed ; Animals ; Aorta ; drug effects ; metabolism ; Dietary Fats ; adverse effects ; Drugs, Chinese Herbal ; pharmacology ; Intercellular Adhesion Molecule-1 ; metabolism ; Male ; NF-kappa B ; metabolism ; Rabbits ; Vascular Cell Adhesion Molecule-1 ; metabolism

OBJECTIVE: The stability of ketamine in biological samples was studied under different storage temperature and time. METHODS: The rabbits were given intragastric administration of ketamine with a dose of 150 mg/kg and were sacrificed after 30 minutes. Blood, liver, kidney and brain of the rabbits were stored at room temperature (between 18 degrees C and 24 degrees C) and -20 degrees C. The specimens were analyzed at different times by GC-MS and GC-NPD. RESULTS: At -20 degrees C, the concentration of ketamine decreased in all of samples (P < 0.05) within 30 days. The concentration of ketamine increased in all of samples stored at room temperature after 5 days(P < 0.05). CONCLUSION The stability of ketamine in biological samples stored at -20 degrees C was better than that at room temperature. The samples suspected containing ketamine should be stored at -20 degrees C and should be tested as soon as possible.
Animals ; Brain/metabolism* ; Cryopreservation ; Disease Models, Animal ; Drug Stability ; Female ; Forensic Toxicology ; Gas Chromatography-Mass Spectrometry/methods* ; Hydrogen-Ion Concentration ; Ketamine/poisoning* ; Kidney/metabolism* ; Liver/metabolism* ; Male ; Rabbits ; Specimen Handling/methods* ; Temperature ; Time Factors

BACKGROUNDPremature ventricular contraction (PVC) is one of the most common kinds of arrhythmias for which the treatment falls into dilemma. Previous clinical application showed that the traditional Chinese Medicine Shensongyangxin (SSYX) capsule is efficacious for the treatment of PVCs. This randomized clinical trial aimed to further evaluate the efficacy and safety of SSYX capsule on treating PVC.

METHODSThe subjects who had frequent PVCs with or without organic heart disease and normal cardiac function were enrolled in the study. The primary endpoint was the change of PVC numbers after eight-week medication with SSYX capsule. The secondary endpoints included change of clinical symptoms related to PVCs and the safety evaluation of SSYX capsule. Totally 188 PVC patients were randomly enrolled in the non-organic heart disease PVCs trial and orally took either SSYX capsules or analogues (three times per day, 4 capsules one time). A total of 671 PVCs patients were randomly enrolled in the organic heart disease PVCs trial, and orally took either SSYX capsules (three times per day, 4 capsules one time) or mexiletine tablet (three times per day, 150 mg one time). The PVCs were monitored and calculated with 24-hour Holter electrocardiogram. Routine blood, liver and kidney function were tested before and after medication with SSYX capsule.

RESULTSSSYX capsules significantly decreased the PVCs numbers and alleviated the related symptoms in patients with or without organic heart disease. In non-organic heart disease group, SSYX capsules and the placebos decreased the PVCs from 12,561.34 ± 9,777.93 to 4,806.87 ± 6,507.17, and 12,605.69 ± 8,736.34 to 10,364.94 ± 9,903.41, respectively. The total effective rate was 74.2% and 28.9% in SSYX and placebo groups (P < 0.001). In organic heart disease group, SSYX capsule and mexiletine decreased the PVCs from 8,641.01 ± 8,923.57 to 3,853.68 ± 7,096.42, 8,621.61 ± 8,367.74 to 5,648.29 ± 8,667.38, respectively. The total effective rate was 65.8% and 50.7% in SSYX and mexiletine groups (P < 0.001). In addition, SSYX capsule significantly alleviated PVCs-related symptoms such as palpitations, chest tightness, insomnia, fatigue, and night sweats. No adverse cardiac events were observed except some slight gastrointestinal side effects during the study.

CONCLUSIONSCompared with placebo or mexiletine, SSYX capsules have significant therapeutic efficacy in reducing PVCs numbers and alleviate PVCs-related symptoms.

Capsules ; therapeutic use ; Double-Blind Method ; Drugs, Chinese Herbal ; therapeutic use ; Humans ; Medicine, Chinese Traditional ; Ventricular Premature Complexes ; drug therapy