To adapt to the new medical enviroment and aiming at the problems existing in clinical teaching,the author explores the value and feasibility of new cultivation mode and its concrete methods to make clininal educatinon play more inportont role in medical education and medical elite cultivation

Objective To study low frequency ultrasound combined with microbubbles to promote the transfection of P53 gene in human prostate cancer cells mediated by liposome.Methods Ultrasound equipment was used with a frequency of 21 kHz and intensity was 46 mW/cm2 and the working time was controlled at 20% (i.e.,2 s “on”time and 8 s “off”time)lasting 5 minutes.The human prostate cancer cell line PC-3 suspension was prepared,the cell concentration was adjusted to 1 × 10 5 cell/ ml,and cells were divided into 8 groups:control group,single microbubbles group,single ultrasound group,ultrasound combined with microbubbles group,single liposome group,liposome combined with microbubbles group, liposome combined with ultrasound group,liposome combined with ultrasound and microbubbles group. Each microbubbles group was added SonoVue 200 μl and the wild type P53 plasmid,plasmid∶liposome is 1 ∶2.At 24 hours after irradiation,Western-blot and RT-PCR were used to detect the gene transfection efficiency,CCK-8 was used to detect cell proliferation,then cell survival rate was calculated,cell apoptosis was detected by flow cytometry.Results After transfection,compared with single liposome group and control group,liposomes combined with ultrasound and microbubbles group can significantly improve expression of the human wild type p53 gene and protein (P <0.001).After transfection,the apoptosis rate of human prostate cancer PC-3 cells in liposomes combined with ultrasound and microbubbles group was significantly higher than that of the liposome group and control group (P <0.001).And after transfection, cell survival rate of liposome combined with ultrasound and microbubble group decreased significantly than those of single liposome group and control group (P <0.001).Conclusions Low frequency and low energy ultrasound combined with microbubbles can promote the transfection of human wild-type P53 gene mediated by liposome.

Objective This study is to identify the presence of superoxide anion-generating system in human lens epithelial cells using arachidonic acid (AA) as the stimulator.Design Experimental study.Participants Human lens epithelial cells B3 (HLE B3). Methods Confluent human lens epithelial cells (HLE B3) were subjected to stimulation by AA and its derivatives.The generation of su- peroxide anion was quantified with a luminometer (LumiStar BMG) immediately upon AA and its derivatives addition.Cells preloaded with superoxide dismutase (SOD) or mannitol were used as negative controls and cells mixed with 3% ethanol (solvent for AA) were used as baseline.Cells were preloaded with inhibitors 30 minutes before luminometer measurement.A time-and concentration-depen- dent study on the AA-stimulated activation of mitogen-activated protein kinase (MAPK) was carried out using western blot analysis. Main Outcome Measures Superoxide generation,phosphorylation of MAPK.Results AA at dosage of 30-150 mM proportionally in- duced luminescence in HLE B3 cells,but was ineffective in cells preloaded with SOD or mannitol.DPI,a non-specific NADPH oxidase inhibitor eliminated AA-induced superoxide anion generation partially.Leinoleic acid,stearic acid,eicosa-11Z,14Z,17Z-trienoic acid (20:3) and eicosa-11Z,14Z-dienoic acid (20:2) were ineffective.The generation of superoxide anion was not contributed by cyclooxyge- nase or the cytochrome p450 pathway since indomethacin (inhibitor for cyclooxygenase) or ketoconazole (inhibitor for cytochrome p450) could not eradicate the stimulatory effect of AA.While CDC,a specific inhibitor for lipoxygenase could eliminate superoxide generation partially.The specific inhibitor for 5-lipoxygenase AA861 completely blocked the generation of superoxide anion.Western blot analysis of the cell lysate showed that AA at the concentrations of 30-150 mM progressively activated ERK and JNK.They were transiently ac- tivated between 2.5-30 minutes.The activations of ERK and JNK were dose-dependent and time-dependent.Conclusions Inhibition of superioxide anion generation may be a new approach to block lens epithelial cell proliferation and post-capsule opacification.

Objective To investigate the effect of heme oxygenase-1 ( HO-1) on brain edema in a rat model of asphyxial cardiac arrest and resuscitation. Methods Forty male SD rats weighing 250-300 g were randomly divided into 4 groups ( n = 10 each) : group I sham operation; group II cardiac arrest (CA); group Ⅰ Hemin (HO-1 inducer) and group IV SnPP (HO-1 inhibitor). Asphyxial cardiac arrest and resuscitation were performed in CA, Hemin and SnPP groups (group Ⅰ ,Ⅲ, Ⅳ) . Hemin and SnPP groups received hemin 15 mg/kg intraperitoneally (IP) at 12 h before CA and SnPP K 30 μmol/kg IP at 1 h before CA respectively. The animals were sacrificed at 1 and 6 h after recovery of spontaneous circulation (ROSC) . The water content of the cortex, hippocampus and brain stem and the expression of HO-1 and aquaporin-4 (AQP4) mRNA in cortex and hippocampus ( by RT-PCR) were determined. Results Water content of cortex and hippocampus was significantly higher at 1 h after ROSC in CA and SnPP groups than in sham operation group and was significantly lower in Hemin group than in CA group. There was no significant difference in water content of brain stem at 1 and 6 h after ROSC among all 4 groups. The expression of AQP4 mRNA was significantly higher in cortex and hippocampus at 1 h after ROSC in CA and SnPP groups than in sham operation group and was significantly lower in Hemin group than in CA group.Conclusion HO-1 can reduce brain water content at early stage after cardiac arrest and resuscitation by regulating the expression of AQP4.

Congresses as Topic ; Rhinitis ; classification ; immunology ; microbiology

ObjectiveTo observe the effect of muscle stimulating instrument on patients with spasticity of lower limbs after surgical treatment.Methods49 adults with spasticity of lower limbs after surgical treatment were divided into the treatment group (21 cases) and the control group (28 cases). Patients of two groups were treated with routien rehabilitation training, but muscle stimulating instruments treatment was added to patients of the treatment group. The muscle strength and motor ability of patients of two groups were followed up and compared.ResultsThe muscle strength and motor ability of the treatment group were better than that of the control group during follow-up period (P<0.05,P<0.01).ConclusionMuscle stimulating instrument can accelerate the recovery of muscle strength and motor ability in adults with spasticity of lower limbs after surgical treatment.

10.3969/j.issn.2095-4344.2013.24.026

Objective To explore the effect of different temperature of the forced- air warming system on the prevention of hypothermia during laparotomy of infants. Methods A total of 60 infants undergoing laparotomy under general anesthesia were recruited and divided into three groups by random digits table method with 20 cases each according to admitting time; when used the force- air warming system intraoperatively, the three groups were respectively setting on 45℃(automatic adjustment for 43 ℃ after 45 minutes), 43 ℃ and 38 ℃.The core temperature were respectively recorded before anesthesia and 15, 30, 45, 60 minutes after anesthesia (every 30 minutes after 1 hour).The hypothermia incidence and anesthesia recovery conditions were recorded simultaneously. Results There was no significant difference on the core temperature among three groups before anesthesia (P > 0.05). 30 minutes after the anesthetic, the core temperature of 45 ℃ group was (36.31±0.20) ℃,43 ℃ group was (36.32±0.24) ℃ and 38 ℃ group was (36.08±0.21) ℃.The differences among three groups was statistically significant (F=8.12, P < 0.01), but there was no significant difference between 45 ℃ group and 43 ℃ group (P > 0.05). 60 minutes after the anesthetic, the core temperature of 45 ℃ group was (36.39±0.26) ℃,43 ℃ group was (36.19±0.22) ℃ and 38 ℃ group was (35.92±0.15) ℃. The differences among three groups was statistically significant(F=25.19, P<0.01).The hypothermia incidence of 45℃group, 43℃ group,38℃ group was 10.0%(2/20), 25.0%(5/20), 50.0%(10/20)respectively and the differences among three groups was statistically significant( χ2=8.04, P<0.05). The time to complete consciousness of 45 ℃ group was (15.40±5.09) minutes,43 ℃ group was (19.80±4.10) minutes and 38 ℃ group was (22.00±4.36) minutes. The differences among three groups was statistically significant (F=10.96, P<0.01). The time to tracheal extubation of 45 ℃ group was (18.10±5.97) minutes, 43 ℃ group was (21.85±4.02) minutes and 38 ℃ group was (24.90±5.54) minutes.The differences among three groups was statistically significant (F=9.83, P<0.01). Conclusions The forced-air warming system can increase the infants′peripheral tissue heat content and reduce the heat losing.So that it will help decrease the intraoperative hypothermia incidence and shorten the anesthesia recovery period.Meanwhile the higher temperature of the forced-air warming system is setted ,the better effect it is.

Objective To compare the clinical data of patients with chronic hepatitis B virus (HBV) carriers and chronic hepatitis B so that to provide pathological evidence for management of chronic HBV carriers with different alanine aminotransferase (ALT) levels.MethodsLiver biopsies were performed in totally of 292 cases of chronic HBV infection.The subjects were divided into HBV carrier group (G0-G1 and S0-S1) and hepatitis group (G＞ 1 and/or S＞ 1) according to the pathological diagnosis. The relationships between different age subgroups, different ALT level subgroups and pathological diagnosis were analyzed. Meanwhile,other clinical,biochemical,and iconographic indexes which were possibly related to the pathology diagnosis were compared. The multivariate analysis was done by Logistic regression equation (withdrawal method, maximum likelihood method) to definite the independent influencing factors of pathologically diagnosed with chronic HBV carrier.ResultsAmong the 292 patients,140 (47.9％) were pathologically diagnosed with chronic HBV carries and 152 (52.1％)were chronic hepatitis B. There were statistical differences between ≤35 years group and 36-40 years,＞40 years group (x2 =3.936 and 8.534,respectively; P =0.047 and 0.003,respectively). There were statistical difference among patients with ALT＜0.5×upper limit normal (ULN),(0.5-1.0) ×ULN,(1.1-1.5) ×ULN,(1.6-2.0) ×ULN and ＞2.0 × ULN (x2 =55.314,P＜0.01),while there was no significant difference between (1.1-1.5) × ULN and ＞2.0 × ULN (x2 =3.810,P=0.051). Multivariate analysis indicated that course of disease,alcohol consumption,ALT,HBV DNA level and the surface of liver (smooth or not smooth)detected by ultrasonography were independent influencing factors of pathological diagnosis of chronic HBV carriers (OR =0.995,0.224,0.516,1.308 and 0.270,respectively; P=0.005,0.007,0.000,0.025 and 0.001,respectively).ConclusionLiver biopsy is much meaningful in patients with age ＞35 years old and ALT (1-2)× ULN.

N-carbamoylase is a part of hydantoinase operon which can transform N-carbamoylamino acid to corresponding ammo acids. The L-N-carbamoylase of Arthrobacter BT801, codied by the HyuC gene, is the rate-limiting and the only stereoselective enzyme. HyuC DNA fragment was amplified by PCR from the plasmid of pUC18-169. The target fragment was introduced into pPIC3. 5K plasmid to construct the pPIC3. 5K-hyuC expressing vector which was then transduced into Pichia pastoris GS115 cells after being linearized by BglⅡ digestion. Multi-copies insertion transformants were screened on G418 plates. The recombinant protein was proved to have biological activity of hydrolyzing N-carbamoylphenylalanine into phenylalanine through enzyme activity determination.