Diabetic Nephropathies ; drug therapy ; Glucosides ; pharmacology ; therapeutic use ; Humans ; Paeonia ; chemistry ; Phytotherapy

Objective To investigate the mastery degree of GCP (Good Clinical Practice) in Guang’anmen Hospital of China Academy of Chinese Medical Sciences (hereinafter referred to as“Guang’anmen Hospital”). Methods Totally 338 clinical researchers, covering 18 clinical professional sections and medical clinical trial institutions, in Guang’anmen Hospital with both qualification of clinical trials and GCP certificate were investigated in 23th-27th June 2014. The investigation was in the written answer sheet form, including four noun explanations and 36 multiple choice questions, which were related to GCP. Results 338 researchers could accurately explain the concepts of 4 nouns, and 57 (16.86%) researchers could do all multiple choice questions correctly. 141 (41.72%) researchers chose 1 wrong answer and 89 (26.33%) researchers chose 2 wrong answers. 39 (11.54%) researchers failed to answer 3-4 questions. Only 12 (3.55%) researchers failed to answer 5 or more questions. Conclusion The mastery degree of clinical researchers with qualification of clinical trials and GCP certificate in Guang’anmen Hospital is satisfied. In the future, pertinence and diverse training should be strengthened, in order to improve the overall level of the mastery degree of GCP.

Objective To investigate the effects of early estrogen replacement therapy(ERT) with different doses on aortic endothelial senescence and its possible mechanism. Methods Twenty- eight healthy New Zealand white female rabbits were randomized into four groups: group A(n=7), in sham operation;group B(n=7), ovariectomized;group C(n=7), ovariectomized and in low-dose ERT(estradiol benzoate 200 μg,im, QOD) ;and group D(n=7), ovariectomized and in high-dose ERT (estradiol benzoate 1000 μg,im,QOD). All rabbits were fed with high fat diet and the rabbits in group C and group D were given ERT from the 7th day after the operation. Blood samples taken via ear central artery before the operation and after the high fat diet for 4 and 12 weeks were used to determine the concentrations of serum estradiol(E2), total cholesterol(TC), low density lipoprotein cholesterol (LDL-C), high density lipoprotein cholesterol (HDL-C), trigtyceride (TG) and asymmetric dimethylarginine(ADMA). After 12 weeks, the aorta was separated for histopathologic analysis and the areas of senescent endothelium and atherosclerotic plaque were calculated.Results (1)After ovariectomized, the concentration of serum estradiol(E2) was significantly lower in group B than in other groups (all P<0.05). Twelve weeks later, the level of E2in was higher group D than in group A and C(both P<0. 05), but there was no significant difference between group C and group A (P< 0. 05). (2)After the high-fat diet for 4 and 12 weeks, there was a notable increase of TC, LDL-C, TG and HDL-C levels in rabbits of each group(all P<0. 01). After 12 weeks, the levels of TC and LDL- C were remarkably higher in group B than in other groups(all P<0. 01), however, the levels of TG and HDL-C were lower in group B than in other groups(all P<0. 01). The concentrations of TC and LDL-C in group C and group D were lower than those in group A (all P<0.01), but the differences of the levels of TG and HDL-C in group C and group D were not significant in comparison with group A (all P>0. 05). There were no significant differences in the lipid levels between group C and D (all P> 0. 05). (3)After 12-week high fat diet, there were a striking increases of serum ADMA level in all four groups (all P>0. 05). The level of ADMA was higher in group B than in other groups (P< 0. 05) andlower in group C and D than that in group A (both P<0. 05). There was no significant difference between group C and D (P<0. 05). (4)12 weeks later, the areas of senescent endothelium and atherosclerotic plaque in group B were significantly larger than those in other groups (all P< 0. 01), and the areas were smaller in group C and D than those in group A (all P<0. 01), there was no significant difference between group C and group D(both P>0.05). (5)Linear correlation analysis showed that there was an obvious positive correlation between the areas of senescent endothelium and atherosclerotic plaque(r=0. 962, P<0. 01), and both of them were positively correlated with the levels of serum AMDA (r=0. 812,0. 824,0. 755,0. 797,0. 749,0. 727), TC (r=0. 812,0. 824) and LDL-C(r=0. 755,0. 797) (all P<0. 01) and negatively correlated with the concentration of serum E2 (r=-0. 762,-0. 743, both P<0. 01).Conclusions Early ERT can improve serum lipid metabolism, reduce ADMA level, inhibit vascular endothelial senescence and attenuate atherogenesis. The delaying effect of estrogen on endothelial senescence may be due to its improving lipid metabolism and lowering ADMA level.

To investigate the effects of salvianolic acid B (SA-B) on cardiovascular endothelial cell function and platelet activation during myocardial ischemia-reperfusion in rabbits.

Objective To explore the effect of inhibiting placental growth factor ( PIGF ) by small interfering RNA ( siRNA) on migration, invasion and chemoresistance of human pancreatic cancer cell line PANC1.Methods Three specific siRNAs targeting PIGF (siRNA-PIGF) were designed.PANC1 cells were transfected with siRNA-PIGF by liposome transfection using untransfected cells as blank controls and nonspecific siRNA ( siRNA-NC) transfected cells as negative controls .The PIGF mRNA and protein expression was examined by real-time RT-PCR and ELISA.MTT method was used to assess the inhibition rate of chemotherapeutic reagents on cell proliferation .The abilities of migration and invasion were evaluated by Transwell assay.Results The inhibition rate of PIGF mRNA in PANC1 cells transfected by 3 siRNA-PIGF were (64.38 ±8.92)%, (70.48 ±7.72)% and (81.25 ±6.02)%, which was lowest in siRNA-PIGF-3 transfected cells.The expression of PIGF mRNA in PANC1 cells were decreased by (63.72 ±8.20)%at 24 h after siRNA-PIGF transfection compared with siRNA-NC transfected cells;and the level of PIGF protein in the supernatant of cultured PANC1 cells was lowered by (42.92 ±1.34)% compared with siRNA-NC transfected cells and by (46.25 ±3.64)% compared with untransfected cells at 48h after transfection, which all had significant difference .There was no statistical difference between untransfected and siRNA-NC transfected cells.After 3 ng/L gemicitabine treatment , the inhibition rate of cell proliferation in siRNA-PIGF group was even higher than that in siRNA-NC and untransfected group [(44.35 ±5.05)% vs(34.29 ±3.60)% and (31.01 ±1.08)%;both P<0.05], and no significant difference was not observed after 5-FU and adriamycin treatment.In migration and invasion assay , the number of transmembrane cells from siRNA-PIGF group was 38.1%and 28.2%of that from siRNA-NC group and 40.8% and 36.2% of that from untransfected group , which had statistical difference (all P<0.05).Conclusions PIGF silencing could significantly suppress the migration and invasion of PANC 1 cells and improve the sensitivity to gemicitabine .

OBJECTIVETo constitute a model of B immunoblastic lymphomas in the Hu-PBL-SCID mice.

METHODSThe SCID mice were reconstituted by intraperitoneal injection (i.p.) of 5 x 10(7) human lymphocytes from Epstein-Barr virus (EBV) seronegative individuals. After one week, the SCID mice were inoculated with EBV by i.p. injection, and subjected to the investigation of whether there was any tumor in the abdomen of such SCID mice four weeks later. The characteristics of the found tumor was observed by the methods of Hematoxylin-eosin (HE) stain, immunohistochemical staining and polymerase chain reaction (PCR).

RESULTSCompared with the control groups, all the EBV-infected Hu-PBL-SCID mice had abdominal solid tumors [(32 +/- 12.5) mm3] developed, often located in the liver. HE staining and immunohistochemical staining showed the tumors were human B cell lymphomas. EBV DNA could be detected in the tumors by the PCR.

CONCLUSIONSThe model of B immunoblastic lymphomas in the Hu-PBL-SCID mice is successfully constituted, and may well be useful to the human tumor immunological study.

Animals ; Disease Models, Animal ; Herpesvirus 4, Human ; physiology ; Humans ; Lymphoma, Large-Cell, Immunoblastic ; Mice ; Mice, SCID

To investigate the effects of gambognic acid (GA) on TRAIL-induced apoptosis of cancer cells, human colon HT-29 cancer cells were treated with GA to promote apoptosis. Inhibition of the cell proliferation was measured with MTT assay and cell apoptosis was detected with formation of DNA ladders in agarose gel electrophoresis, and activation of caspase activity. The content of cytosolic reactive oxygen species (ROS) was measured with flow cytometry. The activities of Caspase-3, -8, -9 were detected using spectrophotometric assay. The levels of c-FLIP, CHOP, DR4 and DR5 in cells were tested by Western blot. Combination of GA (1 µg · mL(-1)) and TRAIL (40 ng · mL(-1)) significantly reduced proliferation and increased apoptosis of HT-29 cells over those induced by each agent alone. Percentage of apoptotic cells was increased to 45.5%. GA markedly enhanced the intracellular ROS generation. Expression of CHOP, DR4 and DR5 was up-regulated to 7.38, 5.41, and 4.85 times of the control group, respectively. GA promoted activation of Caspase-3, -8, and -9 by TRAIL (P<0.05). Furthermore, the expression of anti-apoptotic protein c-FLIP was down-regulated to 0.22 ± 0.08 times of the control group. In conclusion, GA sensitizes HT-29 cells to TRAIL-induced apoptosis by promoting ROS-activated ERS pathways, up-regulating of DR4 and DR5, and inhibiting c-FLIP expression.
Apoptosis ; Apoptosis Regulatory Proteins ; metabolism ; Caspases ; metabolism ; Cell Line, Tumor ; Cell Proliferation ; Colonic Neoplasms ; metabolism ; Down-Regulation ; HT29 Cells ; Humans ; Reactive Oxygen Species ; metabolism ; TNF-Related Apoptosis-Inducing Ligand ; pharmacology ; Up-Regulation ; Xanthones ; pharmacology

AIMTo research the spontaneous firing activities during different-frequency stimulation of subthalamic nucleus and microelectrophoresis GABA, Glu and their antagons respectively, approaching the mechanism of DBS in the treatment of Parkinson's disease further.

METHODSUsing extracellular recording to investigate the effect of different-frequency stimulation of STN and microelectrophoresis several drugs on the spontaneous firing activities of the SNr neurons.

RESULTSFor STN stimulation at low frequency, there was no difference on the spontaneous firing activities of SNr neurons between pro-stimulation and meta-stimulation (P > 0.05). With the increasing of stimulation frequency, most of the SNr neurons were inhibited. While during the STN stimulation frequency at high-frequency, the firing rates of inhibited SNr neurons were changed (P < 0.05). Glu had catatonic excitement effect on the SNr neurons, whereas GABA had tonic inhibition effect. 80% of SNr neurons which were inhibited by STN-HFS were not inhibited by STN-HFS on the basis of excitatory effect of BIC.

CONCLUSIONTo treat the motor symptoms of PD, when SIN is selected as the target nucleus, the electrical stimulation with high-frequency should be chosen. It is possible that SIN-HFS modulate the activity of SNr by inhibitory effect of GABA predominantly.

Action Potentials ; physiology ; Animals ; Electric Stimulation ; Electrophoresis ; methods ; Glutamic Acid ; administration & dosage ; pharmacology ; Male ; Neurons ; physiology ; Parkinson Disease ; physiopathology ; therapy ; Rats ; Rats, Sprague-Dawley ; Substantia Nigra ; physiology ; Subthalamic Nucleus ; physiology ; gamma-Aminobutyric Acid ; administration & dosage ; pharmacology

Objective To explore the expression and regulatory mechanism of hypoxia inducible factor-1?(HIF-1?)during fracture healing. Methods Mouse models of tibia fracture healing were established,and callus samples were collected 1,3,7,14,21 and 28 days after fracture.The development of callus and new bone formation were evaluated with roentgenology,Micro-CT and tetracycline double labeling method,and the expression of HIF-1?,vascular endothelial growth factor(VEGF),Runx2 and ALP in callus were detected with RT-PCR,Western blotting and immunohistochemistry.The relationship between HIF-1? and fracture healing was analysed. Results The expression of HIF-1? was detected in cells in the fracture sites as well as in evolved osteoblasts,chondrocytes and osteocytes in early callus under hypoxia.The highest expression rate of HIF-1? achieved on the 7th day after fracture,lasted for about 7 days,then decreased gradually,and returned to intact level on the 28th day after fracture.The expression tendency of VEGF resembled that of HIF-1?.Bone formation activity was more active in early callus,and the callus volume peaked on the 14th day after fracture and decreased gradually.The mineralization of callus mainly took place in the late healing period(14th to 28th day after fracture).Conclusion Cells involved in fracture healing are hypoxia-responsive cells,which express HIF-1?.HIF-1? can regulate cell state and function,and can promote angiogenesis so as to play a crucial role in fracture healing.

Objective To explore the relationship between some single nucleotide polymorphisms (SNP)loci of interferon regulatory factor 6(IRF6)gene,transforming growth faetor-?(TGFA)gene and nonsyndromic cleft lip with or without cleft palate(NSCL/P)in nuclear families consisting of fathers, mothers and affected offspring with NSCL/P from southeast China.Methods Some SNloci of IRF6 and TGFA were detected by applying microarray technology in nuclear families,and then haplotype relative risk (HRR)and transmission disequilibrium test(TDT)were performed.Results There were no significant difference in genotypes and alleles distribution between patients and their parents.The SNP locus——V274I of IRF6 was associated with NSCL/P(HRR:?~2=4.5816,P