Objective To explore the role of hypoxia-inducible factor-1? (HIF-1?) in the expression of telomerase reverse transcriptase (TERT) in the pulmonary arterial smooth muscle cells (PASMCs) under anoxia.Methods HIF-1? and TERT mRNA in the PASMCs were examined by RT-PCR with the presence of HIF-1? inducers (CoCl_2,DFX),inhibitor (FAS,CBZ-LLL) and oligodeoxynucleotides,or under anoxia.HIF-1? protein in the PASMCs was determined by Western blotting.Results Expressions of HIF-1? and TERT mRNA significantly up-regulated under anoxia (0.59?0.07,0.60?0.06) compared with that in control group ( 0.11? 0.02,0.1?0.01,P0.05).Antisense oligodeoxynucleotides invalidated CoCl_2-induced increase in TERT mRNA levels (0.45?0.04) compared with CoCl_2 group (0.95?0.08,P

To study the in vitro anti-angiogenesis effect of three curcumin pigments (curcumin, demethoxycurcumin, bisdemethoxycurcumin). In the study, the inhibitory effect of the three curcumin pigments on proliferation of HUVEC cells induced by OX-LDL and the effect on migration of HUVEC cells were detected. The effect on neovascularization was observed by chorioallantoic membrane (CAM) test. The effect on cell adhesion factors ICAM-1 and VCAM-1 of HUVECs were tested by Real-time RT-PCR. It was found that the three curcumins could inhibit the proliferation of HUVEC cells induced by OX-LDL within the dosage range 4, 8, 16 mg x L(-1), with a dose-dependence. The proliferative effect of curcumins on HUVECs was greater than the other two derivatives (P < 0.01). All of the three curcumin pigments inhibited the migration of HUVEC cells and the angiogenesis of chick chorioallantoic membrane (CAM). The migration inhibition rate of curcumins at middle and high concentrations was greater than the other two (P < 0.01). All of the three curcumin could down-regulate the expression of VEGF and ICAM-1, and curcumins showed more obvious effect in down-regulating VEGF than demethoxycurcumin and bisdemethoxycurcumin(P < 0.01); Bisdemethoxycurcumin showed the most significant effect in down-regulating ICAM-1 (P < 0.01). All of the three showed no remarkable effect on expression of VCAM-1, and only bisdemethoxycurcumin showed the down-regulating effect (P < 0.05). According to the findings, all of the three curcumin pigments could resist angiogenesis by inhibiting proliferation and migration of endothelial cells and down-regulating the expression of VEGF and adhesion molecules ICAM-1.
Angiogenesis Inhibitors ; pharmacology ; Animals ; Cell Movement ; drug effects ; Cells, Cultured ; Chorioallantoic Membrane ; drug effects ; Curcumin ; analogs & derivatives ; pharmacology ; Humans ; Intercellular Adhesion Molecule-1 ; genetics ; RNA, Messenger ; analysis ; Vascular Cell Adhesion Molecule-1 ; genetics ; Vascular Endothelial Growth Factor A ; genetics

To investigate the difference in absorptive of tetrahydropalmatine (THP) and l-tetrahydropalmatine (l-THP) in rat intestine as well as the mechanism of the absorption of THP, in situ single pass perfusion model was used and the concentration of THP in perfusate was determined by HPLC. The absorption rate constant (k(a)) and effective permeability values (P(eff)) of THP had no significant difference (P > 0.05) at concentration of 8, 16 and 32 microg x mL(-1) in perfusion or in four different regions of intestine of rat (duodenum, jejunum, ileum, colon). The absorption of l-THP and THP in jejunum had significant difference (P < 0.05). The k(a) and P(eff) of THP increased obviously when verapamil was co-perfused with THP, while those of l-THP were not influenced by verapamil. The absorption of THP in intestine showed the passive diffusion process, and without a special absorption region. The stereoselective absorption difference may result from stereoselective combination of P-glycoprotein with d-THP.
ATP-Binding Cassette, Sub-Family B, Member 1 ; antagonists & inhibitors ; Animals ; Berberine Alkaloids ; chemistry ; pharmacokinetics ; Colon ; metabolism ; Duodenum ; metabolism ; Female ; Ileum ; metabolism ; Intestinal Absorption ; drug effects ; Jejunum ; metabolism ; Male ; Perfusion ; Permeability ; Rats ; Rats, Sprague-Dawley ; Sex Factors ; Stereoisomerism ; Verapamil ; pharmacology

AIMTo investigate the change of latency and interpeak latency of each component of BAEP (brainstem auditory evoked potential, BAEP) and its correlation with PV/PFV (pontine volume/posterior fossa volume, PV/PFV) ratio in OPCA (olivopontocerebellar atrophy, OPCA).

METHODSWe used Keypoint EMG/EP to determine waves I PL (peak latency, PL), III PL, V PL and I - III IPL (interpeak latency, IPL), III - V IPL, I - V IPL and used 1.5TMR 3D volume rendering software to determine PV (pontine volume, PV), CV(cerebellar volume, CV) and PFV (posterior fossa volume,PFV). Then calculated PV/PFV ratio, CV/PFV ratio and PV/ CV ratio in OPCA group and control group.

RESULTSCompared with control group, in OPCA group wave IIII PL, I - III IPL were significantly elongated (P < 0.05), III - V IPL was significantly shorten (P < 0.05), PV/PFV ratio was significantly decreased (P < 0.01); there was a positive correlation between III-V IPL and PV/PFV ratio (r = 0.83, P < 0.01).

CONCLUSIONIn patients with OPCA, III PL, I - III IPL of BAEP were elongated and III - V IPL of BAEP was shorten. III - V IPL became shorter when the volume of pontine decreased.

Adult ; Case-Control Studies ; Evoked Potentials, Auditory, Brain Stem ; physiology ; Female ; Humans ; Male ; Middle Aged ; Olivopontocerebellar Atrophies ; pathology ; physiopathology ; Pons ; pathology

OBJECTIVETo study the influence of cervical settling method on marginal accuracy of wax patters.

METHODS24 wax patterns were made on the standard cast specimens and round capsules of which twelve patters were made with the method of cervical settling (experiment group), twelve patters were made with the method of wax dripping (control group). Marginal accuracy of patterns were measured before sublation, sublation half an hour and twenty-four hours.

RESULTSThe results revealed that marginal accuracy had significant difference between the cervical settling methods and wax dripping methods (P < 0.05). There was significant difference between the cervical settling methods measured after half an hour and twenty-four hours (P < 0.01). There was also significant difference between the wax dripping methods measured after half an hour and twenty-four hours (P < 0.01).

CONCLUSIONWax patterns with the method of cervical settling can improve marginal accuracy of patterns. The marginal accuracy of wax patters are influenced by the existing time.

Dental Casting Technique ; Dental Marginal Adaptation

Based on the deletion information of high virulent PRRSV genome, 3 oligonucleotide primer were designed and synthesized. Specific and sensitive reverse transcription-PCR (RT-PCR) assays were de-veloped for the detection of high virulent PRRSV. The sensitivity and specificity of RT-PCR assays were evaluated, the results showing that the detection limit of the assay was found to be 0.265 pg of tissue total RNA, and the protocol have no cross-reaction with classical swine fever virus, porcine circovirus type 2,pseudorabies virus, streptococcus, haemophilus parasuis and Escherichia coli. Then 36 cell cultures, two PRRSV live vaccine strains and 184 clinical specimens from 52 farms were tested. Five PRRSV field iso-lates were the high virulent PRRSV; two PRRSV live vaccine strains from normal PRRSV, and 123 speci-mens from 42 farmer were positive (only 1 specimen was normal PRRSV). This RT-PCR method proved to be accurate differential diagnosis of the high virulent PRRSV and normal PRRSV with the characteristics of rapidity, sensitivity and specificity, and has a strong clinical relevance.

Objective To investigate the application prospective of carboxyfullerene C_3 as a radioprotectant or assistant for tumor radiotherapy.Methods Different concentrations of C_3 were incubated with K562 and AHH-1 cell,CCK-8 assay and trypan blue rejection test were performed to examine the influence of C_3 on the cell viability.Annexin V/PI staining and flow cytometry assay were applied to assess the cell cycle and apoptosis after 7-ray irradiation.Results C_3 showed little toxicity to AHH-1 cell with the survival rate over 95% ,but 600 mg/L of C_3 markedly inhibited the growth of K562 cell (82%) .Pretreatment of 100 mg/L C_3 significantly increased the survival rate of AHH-1 cell after 4 Gy irradiation compared with the single radiation group(71.3% vs 90.3%) ,but decreased the apoptosis rate (26.3% vs 12.6%) ,while the survival rate of K562 cell was decreased and the apoptosis rate was elevated with the increase of C_3 concentration.Moreover,the cell cycle analysis revealed the G_2 phase block in AHH-1 cell after radiation exposure was mitigated by C_3 pretreatment,but that in K562 cell was aggravated.Conclusions C_3 has good radioprotective effects on AHH-1 cells.For K562 cell,C_3 could inhibit the cell proliferation,promote the radiation induced apoptosis and aggravate the G_2 phase block.

Adolescent ; Adult ; Aged ; Ankle Injuries ; surgery ; Female ; Foot Injuries ; surgery ; Humans ; Male ; Middle Aged ; Soft Tissue Injuries ; surgery ; Surgical Flaps

OBJECTIVETo compare clinical efficacy between discectomy and discectomy plus Coflex fixation for lumbar disc herniation.

METHODSFrom December 2007 to August 2008, 50 patients (31 males and 19 females) were treated by surgery of discectomy and discectomy plus Coflex fixation. The average age was 52.5 years (range, 30 - 72 years). There were 24 cases in the group of discectomy plus Coflex fixation and 26 cases in the group of discectomy. Preoperative and postoperative visual analogue scales (VAS), Japanese Orthopadic Association (JOA) and Oswestry disability index (ODI) were recorded, as well as radiological index. And use a paired t-test and one-way analysis of variance (one-way ANOVA) statistical method to evaluate the Coflex dynamic stabilization system in value in the treatment of lumbar disc herniation.

RESULTSBoth groups received significant improvement of JOA, ODI and VAS (t = -33.2 - 64.5, P < 0.01), but the group of discectomy was found with deterioration of ODI at last follow-up, 12 months after surgery 6.7 ± 1.5 to 10.2 ± 2.3 (t = -19.3, P < 0.05). The group of discectomy plus Coflex fixation was found with significant increase of height of dorsal intervertebral discs (HD), distance across the two adjacent spinous processes (DS), distance of intervertebral foramina (DIF) and spinal canal area(SA) (t = -34.4 - 4.5, P < 0.05). In contrast, the group of discectomy was found with significant decrease of HD, DS, DIF and SA (t = 3.4 - 52.8, P < 0.05). Coflex fixed group in HD, DIF, DS significant difference with simple discectomy group, with a statistically significant (F = 14.1 - 25.6, P < 0.05).

CONCLUSIONSBoth discectomy and discectomy plus Coflex fixation are apparently effective when treating lumbar disc herniation. Coflex can significantly increase the HD and DIF when used for lumbar disc herniation, and it has positive influence for keeping height of lumbar vertebral space and treating the nerve root symptom of lumbar disc herniation. Discectomy plus Coflex is better than pure discectomy in preventing lumbar degeneration.

Adult ; Aged ; Female ; Humans ; Internal Fixators ; Intervertebral Disc Displacement ; surgery ; Lumbar Vertebrae ; surgery ; Male ; Middle Aged ; Treatment Outcome

BACKGROUNDUncontrolled cell division is one of the hallmarks of tumor growth. Researches have been focused on numerous molecules involved in this process. Cyclins are critical regulatory proteins of cell cycle progression and/or transcription. The present study aimed to investigate the anti-proliferative effect of cyclin L2, and to define its growth regulatory mechanisms using human lung adenocarcinoma cell line A549.

METHODSHuman cyclin L2 was transfected into human lung adenocarcinoma cells (A549 cell), and was expressed in a mammalian expression vector pcDNA3.1. The effects and mechanisms of the cyclin L2 in cell growth, cell cycle analysis and apoptosis were studied by 3-(4,5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT), flow cytometry or Western blot, respectively.

RESULTSOverexpression of cyclin L2 inhibited the growth of A549 cells. Cell cycle analysis in cells transfected with pCCNL2 revealed an increment in proportion in G0/G1 phase ((68.07 +/- 4.2)%) in contrast to (60.39 +/- 2.82)% of the cells transfected with mock vector. Apoptosis occurred in (7.25 +/- 0.98)% cells transfected with pCCNL2, as compared with (1.25 +/- 0.21)% of the mock vector control group. Cyclin L2-induced-G0/G1 arrest and apoptosis involved upregulation of caspase-3 and downregulation of Bcl-2 and survivin.

CONCLUSIONThe results indicate that overexpression of cyclin L2 protein may promote efficient growth inhibition of human lung adenocarcinoma cells by inducing G0/G1 cell cycle arrest and apoptosis.

Apoptosis ; Caspase 3 ; biosynthesis ; Cell Cycle ; Cell Line, Tumor ; Cyclins ; genetics ; physiology ; Humans ; Lung Neoplasms ; pathology ; Proto-Oncogene Proteins c-bcl-2 ; antagonists & inhibitors ; Transcription Factors ; genetics ; physiology ; Transfection