Objective To determine whether ultrasound mediated microbubble destruction could increase naked human wild type p53 plasmid delivery to ovarian cancer in rats by intratumoral injection. Methods Forty rats with ovarian cancer successfully induced by chemical intoxicant were divided into four groups.The wild type p53 gene was cloned into a high expressing efficiency eukaryotic plasmid pcDNA 3.1+ and the mixture of naked human wild type p53 plasmid was carefully injected directly into the center of ovarian neoplasms with or without echo contrast agent, and it was exposed to ultrasound for twenty minutes once a week.Two months after p53 gene transfection, semiquantitative RT-PCR was used to detect wild type p53 mRNA expression and western blot was to evaluate p53 protein expression level. Results Recombinated eukaryotic expression plasmid DNA encoding pcDNA 3.1+ /p53 was successfully constructed and confirmed by sequence analyzing and endonuclease cutting. The ovarian cancer model of rat was obtained by exposure to intoxicant. Expression of human wild type p53 mRNA was significantly higher in rats transfected with wild type p53 plasmid by means of ultrasound and contrast agent than others(P

Acute Disease ; Child ; Gene Expression Profiling ; Humans ; Leukemia ; diagnosis ; genetics ; therapy

Objective To study the effect of different palatal vault shape on the stress distribution of edentulous maxilla surface.Methods Using three-dimensional finite element method(3-D FED),we simulated five bite states,putting five loading conditions on 3-D FED model of maxillary edentulous with three different palatal vault shapes.Results We obtained the stress value and multicolor picture of 3-D FE model.Labial frenal notch zone and alveolar ridge zone were the stress concentration zone of the maxillary edentulous with three different palatal vault shapes under five kinds of loading conditions.The deep palatal vault maxillary edentulous almost had the maximum stress values in labial frenal notch zone,palate zone and alveolar ridge zone,and upper edentulous jaw with palatal torus was the minimum.Conclusion Palatal vault shape can change the stress value of edentulous maxilla surface,but cannot change the stress distribution.Loading condition can change the stress distribution and stress value.

Mean flow rate (MFR) ,vocal intensity (I) ,pitch frequency(P) ,pitch perturbation (PP) and vocal efficiency (I/F) were measured simutaneously during sustained phonation of the vowel [i] at wide range levels of intensities in 158 nor mal subjects and the same parameters were measured at moderate vocal intensity level in 136 patients with various kinds of laryngeal diseases. PP indicates the amount of irregular vibration of vocal folds and I/F represents efficient transition from MFR to acoustic energy. When phonation at different intensities ,MFR,P and I/F increase but PP decreases when I increases. At moderate intensity phonation, there are positive correlations between MFR and I,and between I and P but negative correlation between MFR and I/F,between I(male) and PP,and between P(fe-male) and PP.Most of the patients with laryngeal diseases showed higher MFR and PP but lower I/F than normal subjects.MFR, PP and I/F could be used as objective and quantitative parameters with I and P as reference parameters in assessing laryngeal function of phonation.

Background Retinal Müller cells can offer nutrient and maintain the normal structure of retina.Researches showed that the abnormality of Müiller cells leads to retinal vascular disease.To explore the effect of high glaucoma on retinal Müller cells is of a very important significance for the study on diabetic retinopathy (DR).Objective This study was to investigate the effects of different concentrations of glucose on retinal Müller cells in vitro.Methods Retinal tissue was isolated from 1 10-day-oM clean SD rat.Mtiller cells were cultured by explant culture method and passaged in DMEM containing 20％ fetal bovine serum.The third generation of cells were obtained and identified using glial fibrillary acidic protein (GFAP) staning.Then,5.5,30.0 and 40.0 mmol/L glucose were added into the culture medium for 4 days respectively.The proliferation (A570) of Müller cells was detected by MTT,and apoptosis rate of Müller cells was calculated by flow cytometer to evaluate the effects of 5.5,30.0 and 40.0 mmol/L glucose to cell vitality.Results Cultured and passaged cells grew well with the spindle shape.The positive reactive cells were ＞95％ for GFAP.The A570 value of Müller cells was 0.24±0.01,0.21±0.03 and 0.20±0.02 in 5.5,30.0 and 40.0 mmol/L glucose group respectively,showing a significant difference among the three groups(F=6.755,P＜0.05).Compared with 5.5 mmol/L glucose group,As70 values were significantly lower in 30.0and 40.0 mmol/L glucose group (q =0.645,0.486,P ＜ 0.05).Apoptosis rates of Miiller cells were (26.40 ±0.25)％,(30.19±0.16)％ and (36.23±0.19)％ in 5.5,30.0 and 40.0mmol/L glucose groups,with a significant difference among them (F =294.530,P＜0.05),and those in 30.0 and 40.0 mmol/L glucose groups were significantly reduced in comparison with 40.0 mmol/L glucose group (q =0.754,0.484,P ＜ 0.05).Conclusions High concentration of glucose inhibits the viability and promote the apoptosis of retinal Müller cells at a concentrationdependent manner.

Adult ; Aged ; Airway Management ; Cyanides ; poisoning ; Female ; Humans ; Male ; Middle Aged ; Poisoning ; nursing ; Young Adult

Acupuncture Therapy ; Child ; Child, Preschool ; Female ; Humans ; Male ; Narcolepsy ; therapy ; Qi

Objective To observe the changes of zymophagy during experimental acute pancreatitis (AP) induced by caerulein.Methods Pancreatic acinar cell line AR42J cells were cultured in 6-well plates till 90％ confluent and then divided into AP group and control group.Caerulein (1 × 10-8 mol/L) was added into AP group to establish AP cell model,and 1640 cell culture medium was added into control group.After caerulein treated for one,four,six,eight,12 and 24 hours,cells and cell culture supernatant were collected.The levels of cytokine interleukin (IL)-1,tumor necrosis factor (TNF)α,trypsinogen activation (TAP) and amylase were measured with enzyme-linked immunosorbent assay (ELISA) method.The expression of LC3 and Beclin1 at mRNA of each group were detected by reverse transcription-polymerase chain reaction (RT-PCR).The LC3B protein level of each group were detected by Western blotting.The changes of autophagosome and zymophagosome were observed by transmission electron microscopy.The difference between AP group and control group was analyzed by analysis of variance.Results The level of IL-1,TNFα,amylase and TAP in cell culture supernatant of control group was (18.83±7.10) pg/mL,(14.20±3.79) pg/mL,(10.03±2.85) U/L and (39.48±8.62) pg/mL,respectively.Those of AP group significantly increased at first hour ((62.13±11.25) pg/mL,F=3.32,P＜0.01 ; (30.98±7.11) pg/mL,F=3.05,P＜0.05; (25.06±6.82) U/L,F=2.90,P＜0.05 and (128.51± 18.30) pg/mL),F=2.62,P＜0.01,at fourth or sixth hour reached peak (IL-1 at fourth hour:(71.96± 15.82) pg/mL,F=7.25,P＜0.01;TNFα at sixth hour:(39.92±8.94) pg/mL,F=4.93,P＜0.05; amylase at fourth hour:(28.83 ± 8.31) U/L,F=2.06,P＜0.05; TAP at fourth hour:(146.29± 29.36) pg/mL,F=0.14,P＜0.01) and then gradually decreased.At fourth and sixth hour,the expression of LC3 at mRNA level in AP group was 3.18±0.82,1.71±0.14,respectively,while the expression of Beclin-1 rnRNA at first,fourth hour was 2.44±0.34 and 4.13±0.30,all of them were significantly increased compared with those of control group (0.21±0.04 and 0.30±0.08,LC3 mRNA F=0.79、0.06; Beclin mRNA F=2.31、0.36,all P＜ 0.05).There were no significant differences at other time points.The numbers of autophagosome and zymophagosome of AP group were significantly higher than those of control group under transmission electron microscopy.Conclusion Zymophagy occurred during AP cell model induced by caerulein,which suggested that zymophagy might involve in the mechanism of AP.

Background:Damage of intestinal mucosal barrier is a key factor in the development and progress of acute pancreatitis(AP),and is closely related with the prognosis of the disease. Aims:To investigate the protective effect and possible mechanism of mucoprotective agent teprenone on intestinal mucosal barrier in rats with experimental AP. Methods:Forty-five adult male Sprague-Dawley rats were randomly divided into normal control group(n = 5),AP model group(n = 20)and teprenone treated group(n = 20). AP model was established by subcutaneous injection of cerulein at abdominal wall. Rats in treated group were intervened with teprenone intragastrically before and after model establishment. ELISA was used for measurement of serum interleukin-1(IL-1),IL-6,tumor necrosis factor-α(TNF-α)and amylase;histopathological and ultrastructural changes of small intestinal mucosa were observed by light microscope and transmission electron microscope;Western blotting was used to detect the expressions of tight junction protein occludin and ZO-1. Results:Serum levels of IL-1,IL-6,TNF-α and amylase in AP model group were significantly higher than those in normal control group(P < 0. 05),accompanied by necrosis and exfoliation of small intestinal villus,widening of intercellular tight junctions and downregulation of occludin and ZO-1 expression. While in teprenone treated group,serum levels of proinflammatory cytokines and amylase were significantly decreased as compared with AP model group(P < 0. 05),the villus of small intestine remained intact,and dense tight junctions were observed. Expressions of occludin and ZO-1 in teprenone treated group were upregulated. Conclusions:Teprenone may protect against intestinal mucosal barrier injury in AP model rats by upregulating tight junction protein expression.

Objective To provide clinical evidence for the optimal acupuncture time in acupuncture treatment of hearing disorder in kids with cerebral palsy.Method Ninety eligible patients were randomized into 3 groups, 30 in each group. Group A was treated for 30 min, group B for 45 min, and group C for 1 h. The hearing improvement was evaluated by using brain-stem auditory evoked potential (BAEP).Result The detection of BAEP showed that the latent and peak periods of wave I,Ⅲ, and V were significantly shortened in the 3 groups after intervention (P<0.01), suggesting that acupuncture can produce a marked efficacy in treating hearing disorder in cerebral palsy. The inter-group comparisons showed that the efficacy of group A was significantly higher than that of group B and C (P<0.05); there was no significant difference in comparing the efficacy between group B and C (P>0.05). It’s indicated that the optimal acupuncture time span should be 30 min.Conclusion With the same acupuncture skills, acupuncture for 30 min can produce a comparatively higher efficacy in treating infantile hearing disorder.