Objective: To investigate the expression of NR4A2(Nurrl) and its splicing variants in gastric cancer, and to evaluate their potential association with the pathogenesis and metastasis of gastric carcinoma. Methods: Alternative Splicing Database (ASD) was used to predict the possible splicing variants of NR4A2; they were identified by gene sequencing technique and their expression was detected by semi-quantitative PCR. The mRNA expression of NR4A2 and its splicing variants in tumor and the corresponding adjacent-tumor tissues (n = 41) were determined by real-time PCR. Meanwhile, immunohistochemisty was employed to examine the protein levels of NR4A2 in the tumor tissues (n = 28). Results: The expression of NR4A2 mRNA was observed in the primary tumor tissues, adjacent tumor tissues, and metastatic tissues. Two splicing variants of NR4A2 were discovered: NR4A2-I and NR4A2-II. The mRNA expression of NR4A2, NR4A2-I, and NR4A2-II in the adjacent-tumor tissue was significantly higher than that in the primary tumor tissue (P = 0.017, P = 0.007, and P = 0.004), and that in the hepatic metastatic tissue was significantly lower than that in the primary tumor tissue (P = 0.001, P = 0.018, P = 0.016). Meanwhile, immunohistochemistry showed that the positive rate of NR4A2 protein was higher in adjacent-tumor tissue than that in both tumor tissues and metastatic tissues, but with no significant differences. Conclusion: At least there are 2 splicing variants of NR4A2 in the gastric tumor, adjacent-tumor, and metastatic tissues; NR4A2 and the novel splicing variants may contribute to the pathogenesis and metastasis of gastric malignancy.

OBJECTIVETo compare the tissue engineered cartilage constructed with chondrocytes derived from auricular and articular cartilage.

METHODSChondrocytes were isolated from porcine auricular and articular cartilage, and BMSCs were obtained from bone marrow aspirate and cultured. Each kind of chondrocytes were resuspended alone or mixed with BMSCs at a ratio of 1:1, and seeded onto PGA/PLA scaffolds to construct tissue engineered cartilage (n = 4). The constructs were cultured for 8 weeks in vitro and then subcutaneously implanted into nude mice for 6 weeks. The differences between chondrocytes monoculture from articular and auricular cartilage or between each of them co-cultured with BMSCs were evaluated by gross view, measurement of thickness and wet weight, histological examinations including H&E, Safranin O, type II collagen, and Ponceau's & Victoria blue staining, and gene expression analysis of cartilage related genes.

RESULTSNo obvious differences were found histologically among the complexes constructed in vitro 8 weeks except for few elastic fibers secreted in the auricular chondrocytes + BMSCs co-culture group. Neo-cartilage is thicker in the groups of articular chondrocytes (38. 1% than the group of auricular chondrocytes, P < 0.05) and articular chondrocytes + BMSCs co-culture (19.3% than the group of auricular chondrocytes + BMSCs, P < 0.05). However, after 6 weeks in vivo the elastic fibers were found positive in the complexes constructed by auricular chondrocytes, and its staining was even stronger and more homogenous in the group of auricular chondrocytes + BMSCs co-culture. The tissues generated by articular chondrocytes alone and co-cultured with BMSCs both formed the characteristic features of three-layer structure of hyaline cartilage and ossified in vivo with significant up-regulation of COL10A1 and MMP-13. To summarize, auricular chondrocytes formed the elastic cartilage while articular chondrocytes formed the hyaline cartilage during the development of tissue engineered cartilage either by monoculture or the co-culture with BMSCs.

CONCLUSIONSThe chondrogenic response of chondrocytes from different cartilage origins demonstrates that an initial chondrocyte and cartilage type recapitulates the same in later tissue-engineered development.

Animals ; Bone Marrow Cells ; cytology ; Cartilage, Articular ; cytology ; Cells, Cultured ; Chondrocytes ; Coculture Techniques ; Ear Auricle ; cytology ; Mesenchymal Stromal Cells ; cytology ; Mice, Nude ; Swine ; Tissue Engineering ; methods ; Tissue Scaffolds

To investigate the immunological enhancement effects of IL-2 with different doses in chicken Shigella vaccine on the duodenum mucosa in Gushi chicken,parental line Gushi chicken were grouped and inoculated with Shigella vaccines including different dose of IL-2.And then the distribution and change of SIgA positive cells and its secretion in the duodenum of the chicken were observed with the immunohistochemistry technology using the Qwin image analysis system.SIgA postive cells were distributed and located primarily in inhesion membrane and enteraden cavity of the duodenum mucosa,additive IL-2 in the Shigella vaccine could increase secretion of SlgA positive cells and strengthen the immunity of the birds,in some degree.The 50 μg IL-2 in the vaccine could display a optimal immunological enhancement effect.

Objective To observe the effects of recombinant human interleukin 11(rhIL-11) on proliferation, migration and invasion of A549 cells, and the mechanism thereof. Methods Final concentrations of 0, 10, 20, 50 and 100μg/L rhIL-11 were added into pulmonary adenocarcinoma A549 cells. The cell proliferation was detected by MTT. The wound-healing, transwell migration assay were used to validate the capability of the migration and invasion of A549 cells. Matrix metallopro-teinases (MMP)-2 and MMP-9 protein expressions were revealed by Western blot assay. Results The proliferation of A549 cells was not significantly changed by rhIL-11. The cell capability to migrate and invade was significantly increased 24 h af-ter treatment with rhIL-11 (P<0.05). The expression levels of MMP-2 and MMP-9 were significantly un-regulated, and which were increased with the increased concentrations of rhIL-11 (P<0.05). Conclusion rhIL-11 can promote the migra-tion and invasion of A549 cells, and the up-regulation of MMP-2 and MMP-9 expression might be one of the mechanisms.

The course of pharmaceutical analysis for pharmacy undergraduate students in National University of Singapore and China Pharmaceutical University was compared, in terms of curriculum, lecturing, assessment and practical teaching.This study provides in-depth analysis by the lecturers from both universities,and can provide reference for teaching reform and course construction of pharmaceutical analysis.

Objective To analyze the prevalence characteristics of fatty liver in young and middle-aged people,and to explore the risk factors of the disease,so as to provide clinical evidence for the prevention and treatment of fatty liver.Methods In 756 young and middle-aged healthy subjects with age≤50 years old,there were 197 fatty liver cases were diagnosed by ultrasonic test during 2015 year.The prevalence of different characteristics in young and middle-aged fatty liver was analyzed,the difference of blood biochemical index between fatty liver and non fatty liver group was compared,and risk factors of fatty liver was explored by binary logistic regression model.Results The total prevalence rate of fatty liver in young and middle-aged people was 26.1%(197/756),among which 33.1% (119/359) were male and 19.6%(78/397) were female,the prevalence rate of male was significantly higher than that of female(χ2=17.833,P<0.05).The prevalence rate of fatty liver increased with age(χ2=33.296,P<0.05),which in 40-50 years old was 37.1%(111/299) and significantly higher than that in 20 years-(15.0%)(24/160) and 30 years-(20.9%)(62/297).Logistic regression analysis on influencing factors of fatty liver prevalence showed that age,sex,body mass index(BMI),drinking,diabetes and fasting blood glucose(FPG),triglyceride(TG),total cholesterol(TC) were closely related to fatty liver(P<0.05),overweight,obesity,drinking,diabetes increased the risk of fatty liver disease.Biochemical indicators(FPG,TG,TC) in fatty liver group were higher than those in non fatty liver group((7.09±1.47) mmol/L vs.(5.14±1.71) mmol/L,(5.98±1.23) mmol/L vs.(4.95±1.42) mmol/L,(2.03±0.45) mmol/L vs.(1.23±0.67) mmol/L,t=271.905,98.866,278.255;P<0.05).Conclusion The prevalence rate of fatty liver in young and middle-aged people is high,and controlling body weight,give up drinking,active treating diabetes,reducing blood glucose and blood lipids can effectively decrease the prevalence of fatty liver in young and middle-aged people.

Study on the effects of Astragali Radix main active flavone calycosin-7-O-β-D-glucoside on Saposhnikoviae Radix main active ingredients prim-O-glucosylcimifugin and cimifugin, a UPLC-MS/MS method for simultaneous determination of prim-O-glucosylcimifugin and cimifugin in rat plasma was established, and the comparative pharmacokinetics of prim-O-glucosylcimifugin and cimifugin after oral administration of prim-O-glucosylcimifugin and calycosin-7-O-β-D-glucoside-prim-O-glucosylcimifugin to rats were carried out, which might be conductive in exploring the rationality of Astragali Radix - Saposhnikoviae Radix herb couple. Twelve male SD rats were divided into two groups. Prim-O-glucosylcimifugin and cimifugin in rat plasma of different time points after oral administration of prim-O-glucosylcimifugin and calycosin-7-O-β-D-glucoside - prim-O-glucosylcimifugin to rats were determinated. And the main pharmacokinetic parameters were investigated using DAS 3. 2. 4. The established method was rapid, accurate and sensitive for simultaneous determination of prim-O-glucosylcimifugin and cimifugin in rat plasma. The analysis was performed on a Waters Acquity BEH C18 column (2.1 mm x 100 mm, 1.7 μm) with the mixture of acetonitrile and 0.1% formic acid/water as mobile phase, and the gradient elution at a flow rate of 0.3 mL x min(-1). The analytes were detected by tandem mass spectrometry with the electrospray ionization (ESI) source and in the multiple reaction monitoring (MRM) mode. Compared with prim-O-glucosylcimifugin group, the AUC(0-t)., and AUC(0-∞) of p-O-glucosylcimifugin as well as the C(max) of cimifugin significantly increased (P < 0.05) in calycosin-7-O-β-D-glucoside-prim-O-glucosylcimifugin group. Calycosin-7-O-β-D-glucoside could enhance the absorption of prim-O-glucosylcimifugin and cimifugin and improve the bioavailability, explaining preliminarily the rationality of Astragali Radix-Saposhnikoviae Radix herb couple.
Animals ; Chromatography, High Pressure Liquid ; Chromones ; blood ; pharmacokinetics ; Drug Interactions ; Drugs, Chinese Herbal ; pharmacokinetics ; Glucosides ; blood ; pharmacology ; Isoflavones ; blood ; pharmacology ; Male ; Monosaccharides ; blood ; pharmacokinetics ; Rats ; Rats, Sprague-Dawley ; Xanthenes ; blood ; pharmacokinetics

Objective In our previous study,we established DNA microarray technology for identification of medical viruses on genus levels and arboviruses on species levels.In this study,we employed these microarrays to determine the pathogen of newly isolated unknown virus in July,2006 from pig brain in Shanxi province.Methods The pathogen isolated from pig brains were inoculated in BHK21 cells.After CPE were observed,the supernatants were collected and RNA was extracted.After reverse transcription and random PCR amplification,the labeled nucleic acids were hybridized with DNA microarrays.Results The hybridization results with medical viruses DNA microarray indicated that the unknown virus belonged to Flavivirus.Combined with epidemiological investigation,we presumed that it might be a kind of arbovirus. Then the labeled specimen were further hybridized with arbovirus DNA microarray and the results confirmed that it was Japanese encephalitis virus(JBV).This coincided with PCR and sequencing analysis.Conclusions The DNA microarray we established previously could be employed to identify unknown viruses.This method provides a new method for determining new viral pathogens.

In order to effectively remove the invalid impurities in Tongan injection, optimize the optimal parameters of the impurity removal technology of liquid mixing process, in this paper, taking Tongan injection as the research object, with the contents of celandine alkali, and sinomenine, solids reduction efficiency, and related substances inspection as the evaluation indexes, the removal of impurities and related substances by the combined process of refrigeration, coction and activated carbon adsorption were investigated, the feasibility of the impurity removal method was definited and the process parameters were optimized. The optimized process parameters were as follows: refrigerated for 36 h, boiled for 15 min, activated carbon dosage of 0.3%, temperature 100 degrees C, adsorption time 10 min. It can effectively remove the tannin, and other impurities, thus ensure the quality and safety of products.
Adsorption ; Charcoal ; chemistry ; Drug Compounding ; instrumentation ; methods ; Drug Contamination ; prevention & control ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Quality Control

To study on the effects of Achyranthes bidentata on Tongsaimai pellets main active ingredients chlorogenic acid, isoliquiritin, harpagoside and glycyrrhizin in rats in vivo pharmacokinetic behaviors, a method for the simultaneous determination of chlorogenic acid, isoliquiritin, harpagoside and liquiritigenin in rat plasma was established by UPLC-MS/MS. The analysis was performed on a waters Acquity BEH C18 column (2.1 mm x 100 mm, 1.7 microm) with the mixture of acetonitrile and 0.1% formic acid/water as mobile phase, and the gradient elution at a flow rate of 0.3 mL x min(-1). The analytes were detected by tandem mass spectrometry with the electrospray ionization (ESI) source and in the multiple reaction monitoring (MRM) mode. It turned out that the analytes of Tongsaimai pellets groups C(max) and AUC(Q-infinity) values were higher than that with A. bidentata group, and the C(max) values of chlorogenic acid had significantly difference (P < 0.05), the AUC(0-infinity) values of chlorogenic acid and glycyrrhizin had significantly difference (P < 0.05); The T(max) and CL values of two groups had no significantly difference. Results showed that the established method was specific, rapid, accurate and sensitive for the studies of Tongsaimai pellets four main active ingredients in rat in vivo pharmacokinetic, and A. bidentata have varying degrees of effects on Tongsaimai pellets four main active ingredients in rat in vivo pharmacokinetic behaviors.
Achyranthes ; chemistry ; Animals ; Chalcone ; administration & dosage ; analogs & derivatives ; blood ; pharmacokinetics ; Chlorogenic Acid ; administration & dosage ; blood ; pharmacokinetics ; Drugs, Chinese Herbal ; administration & dosage ; pharmacokinetics ; Glucosides ; administration & dosage ; blood ; pharmacokinetics ; Glycosides ; administration & dosage ; blood ; pharmacokinetics ; Glycyrrhizic Acid ; administration & dosage ; pharmacokinetics ; Herb-Drug Interactions ; Male ; Pyrans ; administration & dosage ; blood ; pharmacokinetics ; Rats ; Rats, Sprague-Dawley ; Tandem Mass Spectrometry