Objective: To observe the impact of transfection of human granulocyte macrophage colony stimulating factor (hGMCSF) mediated by hydroxyapatite(HA) nanoparticles on the growth of HepG2 cells in order to lay the foundation for further studying gene-modified tumor vaccines of hepatoma in clinic. Methods: The proliferation of HepG2 cells was measured by MTT assay. The plasmids containing hGM-CSF gene were transfected into HepG2 cells mediated by HA nanoparticles. Subsequently, resistant cells were selected by G418 screening. The monoclonal cells were selected by limiting dilution method. RT-PCR was used to identify the integration of hGM-CSF into HepG2 cells and transcription of hGM-CSF. ELISA was performed to detect the level of secreted hGM-CSF in cultured medium and the lasting time. The effect of hGM-CSF on cell cycle and apoptosis were assessed by flow cytometry (FCM) analysis. Results: MTT assay demonstrated that Nano-HA suspension liquid had no significant effects on the growth of HepG2 cells at the concentration below 80 μg/mL. RT-PCR demonstrated the hGM-CSF gene was successfully integrated and steadily expressed in the transfected HepG2 cells. ELISA indicated stable secretion of hGM-CSF from the stable transfected HepG2 cells [mean (216.22 ± 45.78) ng/106 cells per 24 h]. FCM analysis showed that hGM-CSF had no significant effects on the cell cycle and apoptosis of HepG2 cells. Conclusion: hGM-CSF gene could be safely transfected and stably expressed in HepG2 cells mediated by HA nanoparticles. Transfection of hGM-CSF gene mediated by HA nanoparticles has no effects on the growth of HepG2 cells. This study may lay a foundation for the further study of gene-modified hepatoma vaccines.

Objective To observe the effects and mechanism of Ganoderma Lucidum Polysaccharides (GLP) on the activity of superoxide dismutase(SOD), the content of malondialdehyde(MDA) of hippocampus tissue and the learning and memory ability. Methods AD rats induced by A? 25-35 were treated with GLP for 7 days and then observing the Morris water maze to study the learning and memory ability, the content of MDA, and the activity of SOD was measured in the hippocampus tissue with spectrophotometer. Ultrastructural changes of neuron were viewed under transmission electron microscope. Results GLP can distinctly improve the learning and memory ability of A?-injected rats(P

The antitussive activity assay for the root extraction of Disporum cantoniense was carried out with coughing mice induced by ammonia liquor. The results showed that the ethanol and water extractions of D. cantoniense possess strong antitussive activity, and the high dose of the former was better than positive control, and then the constituents of the ethanol extraction were separated and purified by various modern chromatographic techniques. Their structures were identified by physico-chemical properties and spectroscopic data. As a result, eight compounds were isolated and identified as stigmast-4-en-3-one(1), (22E, 24R)-ergosta-5, 7, 22-trien-3beta-ol(2), obtucarbamate A(3), obtucarbamate B(4), neotigogenin(5), azo-2, 2'-bis[Z-(2,3-dihydroxy-4-methyl-5-methoxy) phenyl ethylene] (6),dimethyl {[carbonylbis (azanediyl)] bis( 2-methyl-5, 1-phenylene) j dicarbamate (7) , and quercetin-3-O-pB-D-glucopyranoside(8). All compounds were isolated from this plant for the first time, and the result of bioactivity-directed isolation showed that compounds 3, 4, and 6 had obvious effect on antitussive activity, and compound 6 had the same level as positive control.
Animals ; Antitussive Agents ; chemistry ; isolation & purification ; pharmacology ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; pharmacology ; Ethanol ; chemistry ; Female ; Liliaceae ; chemistry ; Male ; Mice

Objective To investigate the relationship between platelet maximum aggregation rate(MAR),platelet thrombocyt-ocrit(PCT),platelet count(PLT),platelet distribution width(PDW)and mean platelet volume(MPV)with the course of acute cere-bral infarction(ACI)to provide the basis for its clinical early diagnosis and treatment.Methods 107 patients with ACI in our hospi-tal were selected and divided into the great infarction group(infarction size >10 cm3 ),middle infarction group(infarction size 4-10 cm3 )and small infarction group(infarction size<4 cm3 )according to the infarction lesion size by head CT or MRI and the infarction volume calculated by the Pullicino formula(length×width×layer number/2),40 healthy individuals were selected as the healthy control group.MAR,PLT,PDW,MPV and PCT were detected before and after the induction by PLR-06.Results (1 )Compared with the control group,PLT,PCT,PDW and MPV before the induction by PLR-06 in the great infarction group were obviously in-creased(P <0.01);PLT,PCT,PDW and MPV in the middle and small infarction groups were increased(P <0.05).(2)Compared with the control group,MAR after the induction by PLR-06 in each infarction group was increased(P <0.05);PLT had no statisti-cal difference among the groups(P >0.05 );PCT,PDW and MPV in the great and middle infarction groups were increased(P <0.05);PCT,PDW and MPV in the small infarction group had no statistical differences(P >0.05).Conclusion The change of the platelet aggregation rate,number and volume is closely related with the occurrence and development of ACI,monitoring their change has important clinical significance to prevention and treat ACI.

OBJECTIVETo investigate the relationship between the intake of ethanol and sexual function of rats.

METHODSSixty adult male Wistar rats were randomly divided into five groups: the control, 10% , 20% , 30% and 40 ethanol groups, which received. . 9% sodium chloride, 10% , 20% , 30% and 40% ethanol solutions respectively at a dose of 2 ml through gastric tubes once a day. Three months later, we observed the effects of ethanol on the sexual function of the rats by their sexual behaviors, the number of apomorphine-induced penile erections, and the content of testosterone in the serum and nitric oxide synthase ( NOS) in the penis.

RESULTSCompared with the control group, the number of apomorphine-induced penile erections in the 10% and 20% ethanol groups was not inhibited significantly (P > 0.05), but the latent period of mounting and intromission in the 10% ethanol group was prolonged and the sexual behaviors in the 20% ethanol group were inhibited except the latent period of ejaculation. The sexual behaviors and the number of apomorphine-induced penile erections of the 30% and 40% ethanol groups were inhibited significantly (P < 0.05). Testosterone in the serum and NOS activity in the penis of the experimental groups were reduced (Pat < 0.05).

CONCLUSIONAn adequate volume of ethanol does not induce sexual dysfunction in rats, but long term and excessive intake of ethanol may cause penile erectile dysfunction.

Animals ; Dose-Response Relationship, Drug ; Ethanol ; administration & dosage ; pharmacology ; Male ; Nitric Oxide Synthase ; metabolism ; Penile Erection ; drug effects ; Penis ; metabolism ; Random Allocation ; Rats ; Rats, Wistar ; Sexual Behavior, Animal ; drug effects ; Testosterone ; blood

BACKGROUNDVoltage-gated K+ channel (Kv) plays a critical role in the modulation of detrusor contraction. This study was conducted to investigate the expressions of Kv2.1 and Kv2.2 in rat bladder with detrusor hyperreflexia (DH).

METHODSThirty adult female Sprague-Dawley rats (200-220 g) were randomly divided into the control group and the experimental group. The experimental group was subjected to spinal cord injury (SCI). In the controls, the surgical procedure was identical with the exception that dura and spinal cord were transected. Four weeks after SCI, in vivo cystometry and mechanical pulling tests of isolated detrusor strips were performed. mRNA was extracted from the detrusors of normal and DH rats for the detection of expression of Kv2.1 and Kv2.2 by RT-PCR. Differences in expression between normal and overactive detrusors were identified by gel imaging.

RESULTSFourteen rats in the experimental group exhibited uninhibited bladder contraction (>8 cmH2O) before voiding after SCI. One rat died from infection. The frequency of DH in the experimental group was significantly different from that in the control group with or without treatment with 4-aminopyridine (4-AP) (P < 0.05), while the amplitude of DH did not change markedly. The rates of variation of the automatic contractile frequency and amplitude were (66.8 +/- 12.4)% and (42.6 +/- 12.6)% respectively in the control group, and (38.4 +/- 9.8)% and (28.0 +/- 4.6)% respectively in the DH group. 4-AP increased the automatic contractile frequency apart from the automatic contractile amplitude in both the control and DH groups (P < 0.05). 4-AP increased the rate of variation of the automatic contractile frequency more markedly in the control group than in the DH group (P < 0.05). Significant expression of Kv2.2 was not detected in bladders in the control group. Compared to the mRNA levels of beta-actin, the mRNA level of Kv2.1 was 1.26 +/- 0.12 in the control group and 0.66 +/- 0.08 in the DH group. SCI significantly reduced the mRNA level of Kv2.1 in rat bladders with DH (P < 0.05).

CONCLUSIONSOur study showed that the mRNA level of Kv2.1 decreased significantly in rat bladder with DH, which was one of the important pathogenetic mechanisms for DH, and suggested that Kv2.1 might be one of the therapeutic targets for bladder overactivity.

Animals ; Female ; In Vitro Techniques ; Muscle Contraction ; RNA, Messenger ; analysis ; Rats ; Rats, Sprague-Dawley ; Reverse Transcriptase Polymerase Chain Reaction ; Shab Potassium Channels ; genetics ; physiology ; Urinary Bladder ; metabolism ; Urinary Bladder, Overactive ; etiology ; metabolism

AIMTo search for novel antiasthmatic agents.

METHODSCoupling seratrodast (SD), an antiasthmatic drug, with several different types of NO donors including oxatriazoles, N-hydroxyguanidines and furoxans; evaluating the antiasthmatic effects of coupled compounds by determining their inhibitory activity of guinea pig asthma induced by acetylcholine and histamine; and assessing NO releasing ability.

RESULTSNine novel target compounds (I1-9) were synthesized, and their structures were established by IR, NMR, MS and elemental analysis. Preliminary pharmacological test showed that most of the compounds showed high antiasthmatic activities (the latent period of induced asthma was prolonged from 10 s (SD) to 26-62 s), among which 3 compounds (I4, I6, I7) were more potent than SD (P < 0.05, P < 0.01) and released more NO than others. The maximum concentrations (Cmax) of NO-release in vitro were 0.1878, 0.1393 and 0.2473 mg x L(-1), respectively.

CONCLUSIONNO donating-SD derivatives are worthy to be futher investigated.

Acetylcholine ; Animals ; Anti-Asthmatic Agents ; chemical synthesis ; pharmacology ; therapeutic use ; Asthma ; chemically induced ; prevention & control ; Benzoquinones ; chemical synthesis ; pharmacology ; therapeutic use ; Guanidines ; chemistry ; pharmacology ; Guinea Pigs ; Heptanoic Acids ; chemical synthesis ; pharmacology ; therapeutic use ; Histamine ; Nitric Oxide ; metabolism ; Nitric Oxide Donors ; chemistry ; pharmacology ; Oxadiazoles ; chemistry ; pharmacology ; Structure-Activity Relationship

OBJECTIVETo study the dynamic effects of allogenic transplantations with the bladder acellular matrix grafts (BAMG) of rabbits.

METHODSHemi-cystectomies were performed in 25 rabbits, and the defects were repaired with BAMG about half bladder size. The rabbits underwent postoperative assessment of bladder function at 8 weeks, including cystometry, vesical volume, vesical compliance and cystography. The allografts were observed by light microscope and electron microscope at 1, 2, 4, 8, 12, 16 weeks after surgery.

RESULTSMacroscopic observation revealed that BAMG regenerated gradually. All urodynamic results of 8 weeks after surgery were not different statistically as compared with these of preoperation (P > 0.05). Cystography revealed that the morphous of bladder was recovered. Epithelialization and neovascularity occurred accompanied by infiltration of inflammatory cell at 1 week. Smooth muscle cell and stratified epithelium regenerated 2 weeks after grafting. Neural elements formed around smooth muscle bundles as early as 4 weeks. Each component regenerated on the frame of BAMG sequentially. After 16 weeks, it was difficult to delineate the junction between the host bladder and BAMG by histology.

CONCLUSIONAfter allogenic transplantation with rabbits' BAMG, the constitution and function of the allografts regenerate completely and gradually on the frame of BAMG.

Animals ; Extracellular Matrix ; transplantation ; Rabbits ; Regeneration ; Tissue Engineering ; Transplantation, Homologous ; Urinary Bladder ; cytology ; physiology ; transplantation

OBJECTIVETo investigate the effects of alcohol intake on penile structure and function in rats.

METHODSThirty adult male Wistar rats were randomly divided into two groups: control group and alcohol intake group. They were administered with 2 mL of normal saline and 40% alcohol solution respectively through gastric tubes every day. Three months later, the animal model of alcohol intake was evaluated by modified Nayagida's method, and the effects of alcohol on the rats were studied by sexual behavior, the number of apomorphine-induced penile erection, level of testosterone in the sera, and the content of penile smooth muscle.

RESULTSThe scores of animal model of alcohol intake evaluated by Nayagida's method were 0.66 +/- 2.05 in the control group and 9.26 +/- 5.50 in the alcohol intake group (P < 0.05), which indicated that an animal model of alcohol intake was successfully established. Sexual behavior, the number of apomorphine-induced penile erection, testosterone level in the sera, and the content of penile smooth muscle of the alcohol intake group were all statistically different as compared with the control group (P < 0.05).

CONCLUSIONAlcohol intake induces sexual dysfunction in rats, which may be due to the decline of testosterone level in the sera and decline of penile smooth muscle.

Animals ; Ethanol ; adverse effects ; Female ; Male ; Penis ; anatomy & histology ; drug effects ; physiology ; Rats ; Rats, Wistar ; Sexual Behavior, Animal ; Testosterone ; blood

Revolution energy spectral CT had its structural innovation introduced from the aspects of detector,driving system and CT bulb as well as high voltage generator,technical principle analyzed from the aspects of SSF,multi-model iterative reconstruction, cardiac imaging unlimited as well as material separation and quantitative analysis, and clinical application described in diagnoses of liver cancer, cholecystolithiasis and kidney stone, coronary arteriongraphy and metal artifact elimination.It's pointed out Revolution energy spectral CT was a new method for identifying the focal nature,tumor homology and components of inorganic substance as well as analyzing multi material quantitatively and qualitatively.