Objective To analyze the effects of acute depletion of liver-specific insulin signaling on secretion of apolipoprotein B (apoB) and triglyceride (TG).Methods Based on Cre-LoxP principle,a promoter of hepatic tissue specific albumin gene was inserted into upstream of the cre recombinase gene.Albumin-Cre adenovirus (Ad-CRE) and GFP adenovirus (Ad-GFP) were amplified in 293A cells and purified before intravenous administration to mice.After adenovirus infection for 2 days,4 days and 6 days,blood samples from mice were collected and hepatic tissues were frozen.The secretion rates of hepatic newly synthesized apoB and very low density lipoprotein (VLDL)-TG were determined by injection of Triton WR-1339.The levels of plasma cholesterol (TC) and TG were measured.The expressions of insulin receptor and other lipoprotein metabolism related proteins in hepatic tissues were analyzed by Western blot.Results After 2 d,4 d and 6 d of the Ad-CRE injection into mice,insulin receptor expression was reduced by 30.50％ (P＜0.05),60.12％ (P＜ 0.01) and 99.54％ (P＜0.001),and VLDL-TG secretion rate was decreased by 20.43％ (P＜0.05),33.63％ (P＜0.05) and 44.21％ (P＜0.01),respectively.Expressions of sterol regulatory binding proteins 1,fatty acid synthase,and the related proteins of VLDL-formation were decreased,but there were no changes in hepatic secretion of apoB100 and hepatic lipids.The hepatic secretion of apoB48 was increased by 35.07％ (P＜0.05) 6 d after Ad-CRE injection.Conclusions Acute depletion of hepatic insulin receptor might reduce VLDL-TG secretion in manner of time-dependent,and increase the assembly and secretion of smaller apoB-containing lipoproteins in mice liver,which is probably associated with decreased lipogenesis.

AIM: To explore the therapeutic effects of flunarizine combined with aspirin in the treatment of migrainous cerebral infaction. METHODS: 38 cases of patients diagnosed as migrainous cerebral infaction were respectively given flunarizine 10 mg combined with aspirin 100 mg every night for a month. The observed indices included the dysfunction scores of nervous system, the total classes of living ability, and the accumulating rate of platelet and viscosity of plasma before and after the treatment. RESULTS: Before the treatment the dysfunction scores of nervous system and the living ability of the patients were 13.51 ? 4.78 and 3.45 ? 1.13 , and after the treatment the values were 4.34 ? 1.85 and 1.79 ? 0.72 respectively (P

OBJECTIVETo investigate the application of facial liposuction and fat grafting in the remodeling of facial contour.

METHODSFrom Nov. 2008 to Mar. 2014, 49 cases received facial liposuction and fat grafting to improve facial contours. Subcutaneous facial liposuction with tumescent technique and chin fat grafting were performed in all the cases, buccal fat pad excision of fat in 7 cases, the masseter injection of botulinum toxin type A in 9 cases, temporal fat grafting in 25 cases, forehead fat grafting in 15 cases.

RESULTSMarked improvement was achieved in all the patients with stable results during the follow-up period of 6 - 24 months. Complications, such as asymmetric, unsmooth and sagging were retreated with acceptance results.

CONCLUSIONCombination application of liposuction and fat grafting can effectively and easily improve the facial contour with low risk.

Adipose Tissue ; transplantation ; Botulinum Toxins, Type A ; administration & dosage ; Chin ; Face ; surgery ; Forehead ; Humans ; Injections, Intramuscular ; Lipectomy ; adverse effects ; methods ; Masseter Muscle ; Neuromuscular Agents ; administration & dosage

Objective To evaluate the long term effect of video-assisted thoracoscopic thymectomy for myasth,enia gravis and the influence of concomitant thymoma.Methods 47 cases of myasthenia gravis were retrospectively reviewed who had received video-assisted thoracoscopic thymectomy from Apr.2001 through Oct.2009.The patients were separated to two groups with or without thymoma.Influence of oncologic factors was carefully studied.Results There were 20 males and 27 females with a mean age of 36.6 yrs.According to the typing system of the Myasthenia Gravis Foundation of America ( MGFA),the patients belonged to type Ⅰ 18 cases,type Ⅱ a 14 cases,type Ⅱ b 14 cases,and Ⅲa 1 case.22 patients were in the group with thymoma,and the other 25 in the group without thymoma.Until the deadline of follow-up time of Jun.2011,only two cases in non-thymomatous group were lost.Follow-up time was 20 to 122 months,mean 57 months.The complete stable remission rate(CSR),pharmacologic remission(PR),minimal manifestations(MM),worse(W),exacerbation(E) and died of myasthenia gravis(D) in non-thymomatous group were 78.3％,13.0％,4.3％,0,0 and 4.3％.In thymomatous group the values were 50.0％,22.7％,13.6％,4.5％,9.1％ and 0.Conclusion Video-assisted thoarcoscopic thymectomy has a satisfactory long term effect for myasthenia gravis.Thymomatous group has no different in overall effectiveness with that of non-thymomatous group although a probably lower complete stable remission rate is prompted.

AIMTo study the relationship between changes of learning, memory and orphanin FQ (OFQ) in hippocampus of rats in acute hypobaric hypoxia at two different altitude levels of 4 500 m(moderate) and 7 500 m(serious).

METHODSHypobaric chamber, Morris water maze training method and RT-PCR technique were used in the experiment.

RESULTS(1) Compared with the control, the OFQ mRNA expression of hippocampus increased significantly after exposure to acute serious hypobaric hypoxia (8 h/d for 6 d in succession), but obviously decreased after Morris water maze training (6 times/d for 6 d in succession, the latency of place navigation was shortened). (2) After exposure to acute hypobaric hypoxia, the latency of place navigation was markedly elongated, but the OFQ mRNA of hippocampus was significantly higher than that of learning and memory group.

CONCLUSIONThe hippocampal OFQ was involved in the mechanism of decrease in spatial learning and memory induced by acute hypobaric hypoxia.

Altitude ; Animals ; Hippocampus ; metabolism ; Hypoxia ; Maze Learning ; Memory ; Opioid Peptides ; physiology ; RNA, Messenger ; genetics ; Rats ; Rats, Wistar

Objective To explore the feasibility and advantages of establishment of retrograde amnesia (RA) model in mice with such methods as electric shock,anoxia,and anesthesia.Methods Kunming mice were divided into control group and 5 treatment groups,including treatments with electric shock,anoxia,propofol,electric shock+anoxia,electric shock+propofol.At first,every group received the same dark avoidance training to establish the behavior of dark avoidance,then the 5 treatment groups were treated with the methods of 120-180 V electric shock,anoxia within a closed container,intraperitoneal injection of 0.3 mL of propofol,electric shock+anoxia,electric shock+propofol,respectively.Next day,step-in latency (Lat) of mousse were measured with the dark chamber in all groups and changes of dark avoidance behavior were analyzed.Results The Lat in the control group 24 h after dark avoidance training was (111.7+17.2) s.In the treatment groups of electric shock,anoxia,electric shock+anoxia,electric shock+propofol,significantly shortened Lat,which limited to 30 s in some mice 24 hatter corresponding treatment,was observed as compared with that in the control group (P<0.05).Except for the propofol treatment group did not changed obviously,the incidence rate of shortened Lat was 43.8%,45.4%,66.7% and 60% in the electric shock treatment group,anoxia treatment group,electric shock+anoxia treatment group,and electric shock+propofol treatment group,respectively.On the 5th and 8th d,some mice recovered from the shortened Lat.Conclusion RA model can be established successfully in some mice treated with electric shock,anoxia,electric shock+anoxia,electric shock+propofol and the highest modeling rate was found in the electric shock+anoxia treatment group.RA can recover in the later stage in some modeling mice and use of pmplfol alone call not induce RA.

AIMTo search for new artemisinin derivatives with higher immunosuppressive activity.

METHODSTwo kinds of new artemisinin derivatives containing polyethylene glycol group were synthesized from dihydroartemisinin via condensation and esterification. These compounds were assayed for their inhibitory activity on ConA-induced T cell proliferation and LPS-induced B cell proliferation.

RESULTSTwenty three new compounds (2a - 2f, 3a - 3d, 4a - 4f, 6a, 6b and 7a - 7g) were synthesized and identified by 1H NMR and elemental analysis.

CONCLUSIONThese compounds had immunosuppressive activity in vitro. Among them, the symmetrical substituted compound 2 and 6 had higher activity than mono-substituted compound 3, 4 and 7. Especially, compounds 2a - 2f remarkably exhibited higher inhibition in comparison with artemisinin and artesunate.

Animals ; Artemisinins ; chemical synthesis ; pharmacology ; B-Lymphocytes ; drug effects ; Cell Proliferation ; drug effects ; Humans ; Immunosuppressive Agents ; chemical synthesis ; pharmacology ; Molecular Structure ; Polyethylene Glycols ; Sesquiterpenes ; chemical synthesis ; pharmacology ; T-Lymphocytes ; drug effects

BACKGROUNDCockayne syndrome (CS) is a rare human genetic disorder characterized by increased UV sensitivity, developmental abnormalities and premature aging. Cells isolated from individuals with CS have a defect in transcription-coupled DNA repair. Despite the repair defect, there is no any increased risk of spontaneous or UV-induced cancer for CS individuals. The strategy of RNA interfering was used here to explore the potential radiosensitizing and anticancer activity of targeting CS group B (CSB) gene.

METHODSThe vectors encoding CSB-specific siRNAs were constructed by inserting duplex siRNA encoding oligonucleotides into the plasmid P(silencer TM 3.1). The cell lines expressing the CSB-siRNA were generated from HeLa cells transfected with the above vectors. Colony-forming ability was used to assay cell survival. Cell cycle was analyzed by FACScan flow cytometry. The apoptosis was measured by detecting the accumulation of sub-G(1) population as well as by fluorescence staining assay. Reverse transcriptase polymerase chain reaction (RT-PCR) was used to semi-quantify mRNA expression. Protein level was detected by Western blotting analysis.

RESULTSTwo constructs encoding CSB-specific siRNA were generated, both of them resulted in remarkable suppression on CSB expression in HeLa cells, and led to an increased sensitivity to (gamma-ray and UV light. siRNA-mediated silencing of CSB decreased cell proliferation rate, increased spontaneous apoptosis as well as the occurrence of UV- or cisplatin-induced apoptosis by 2 to 3.5 fold. A significant S phase blockage and a remarkable reduction of G(1) population were induced in control HeLa cells at 18 hours after being exposed to 10 J/m(2) of UV light. The S phase blockage was also observed in UV-irradiated CSB-siRNA transfected HeLa cells, but the extent of increased S phase population was lower than that in the UV-irradiated control cells. No or a relative weak reduction on G(1) phase population was observed in UV-irradiated CSB-siRNA transfected HeLa cells. In addition, siRNA-mediated silencing of CSB promoted the elimination of G(2)/M phase cells after UV light radiation.

CONCLUSIONSsiRNA-mediated silencing of CSB causes cells to proliferate more slowly, sensitize cells to genotoxicants, and modify UV radiation-induced cell cycle changes. siRNA-mediated inactivation of CSB could be an attractive strategy for ameliorating cancer therapy, which can be fulfilled via the combination of gene therapy and sensitization of radiotherapy or chemotherapy.

Apoptosis ; radiation effects ; Cell Cycle ; radiation effects ; Cell Proliferation ; radiation effects ; Cisplatin ; pharmacology ; Cockayne Syndrome ; genetics ; Gene Silencing ; Genetic Therapy ; HeLa Cells ; radiation effects ; Humans ; RNA, Small Interfering ; genetics ; Radiation Tolerance ; Ultraviolet Rays

OBJECTIVETo analyze the drug-sensitivity-related genes to anti-tumor drugs navalbine (NVB) and docetaxel (Doc) in four SCLC and six NSCLC cell lines.

METHODSThe sensitivity of 4 SCLC lines and 6 NSCLC lines to NVB and to Doc was determined with MTT test. The expression of 1291 anti-tumor drug sensitivity-related genes in the 10 cell lines was assayed by cDNA macroarray technique, and cluster analysis was performed to find the relationship between the results obtained by the above mentioned two measurements.

RESULTS(1) The anti-tumor effect of NVB on the 10 cell lines was apparently better than that of Doc. (2) The drug sensitivity-related genes in these 10 cell lines showed a more close positive correlation with Doc than that with NVB, whereas more genes showed negative correlation with NVB than that with Doc. But in 6 NSCLC cell lines, more genes showed the same positive or negative correlation with the two drugs. (3) 51 genes in the 10 cell lines showed correlation with Doc or NVB. 13 of them had negative correlation with Doc, 11 of them showed positive correlation. 24 of them showed negative correlation with NVB, 3 of them showed positive correlation. 67 genes in 6 NSCLC cell lines showed a correlation with sensitivity to Doc or NVB, among them 34 had negative correlation with Doc, 4 had positive correlation. 25 genes had negative correlation with NVB, 4 had positive correlation. (4) Rab 1, Rab 3, Rho B, Rho C, Rac 1, Rac 2, Gho GDI beta, CD44, integrin alpha5, integrin alpha6, integrin beta5, vinculin showed to be cytoskeleton-related genes differently expressing in SCLC or NSCLC cell lines.

CONCLUSIONThere is obvious difference in the drug sensitivity-related genes to NVB or Doc between SCLC and NSCLC cell lines.

Antineoplastic Agents ; pharmacology ; Cell Line, Tumor ; Cluster Analysis ; Cytoskeletal Proteins ; metabolism ; Cytoskeleton ; genetics ; Gene Expression Profiling ; Gene Expression Regulation, Neoplastic ; Humans ; Integrin alpha5 ; metabolism ; Lung Neoplasms ; genetics ; metabolism ; pathology ; Oligonucleotide Array Sequence Analysis ; Taxoids ; pharmacology ; Vinblastine ; analogs & derivatives ; pharmacology ; rab1 GTP-Binding Proteins ; metabolism ; rac1 GTP-Binding Protein ; metabolism

OBJECTIVETo investigate the role of extracellular regulated kinase (ERK1/2) pathway in cisplatin-induced apoptosis in human ovarian carcinoma cells.

METHODSCisplatin-induced apoptosis were stained with DAPI and was assessed microscopically in human epithelial adenocarcinoma ovarian cell line SKOV3 cells. ERK activation was determined by Western blotting using an anti-phospho-ERK antibody to detect ERK activity. The effect of PD98059 on ERK activity induced by cisplatin was detected by MTT assay.

RESULTSMarked apoptosis of SKOV3 cells resulted from 48 hours treatment with 20 microg/mL cisplatin. Strong activation of ERK was led to by 15 microg/mL cisplatin. Dose response and time course of cisplatin induced apoptosis in SKOV3 cells. Cisplatin-induced ERK activation occurred at 12 hours and increased to highest induction at 24 hours by Western blotting. The effect of PD 98059 on ERK activity induced by cisplatin at the concentration of 100 micromol/L PD 98059. Statistically significant decreased in cell survival were observed with 100 micromol/L PD 98059 at 15 and 20 microg/mL cisplatin (P < 0.05).

CONCLUSIONSCisplatin activates the ERK signaling pathway in ovarian cancer cell line SKOV3. Inhibition of ERK activity enhances sensitivity to cisplatin cytotoxity in ovarian cancer cell line SKOV3. Evaluation of ERK activity could be useful in predicting which ovarian cancer will response most favorably to cisplatin therapy.

Adenocarcinoma ; enzymology ; pathology ; Antineoplastic Agents ; pharmacology ; Apoptosis ; drug effects ; Cell Line, Tumor ; Cell Survival ; drug effects ; Cisplatin ; pharmacology ; Enzyme Activation ; drug effects ; Female ; Flavonoids ; pharmacology ; Humans ; Mitogen-Activated Protein Kinase 3 ; Mitogen-Activated Protein Kinases ; metabolism ; Ovarian Neoplasms ; enzymology ; pathology ; Signal Transduction