The purpose of this paper is to investigate the reversal effect of small interfering RNA (siRNA) targeting MDR1 and MDR3 genes on the resistance of MCF-7/ADR cells to adriamycin. siRNA plasmid vector targeting MDR1 and MDR3 genes was transfected into MCF-7/ADR cells, and then was stained with Annexin-V FITC (fluorescein isothiocyanate conjugated) to detect the early stage cell apoptosis by flow cytometry (FCM). 50% inhibition concentration (IC50) of adriamycin for MCF-7/ADR cells was determined by MTT method. MDR1 and MDR3 mRNA was assessed by RT-PCR. Treatment of MCF-7/ADR cells with the two kinds of siRNAs resulted in a reversal of adriamycin resistance of MDR to different extents. 1) The apoptosis efficiency of MDR1 and MDR3 siRNA vector after transfection was (18.21+/-1.65) % and (9.07+/-2.16) % respectively (P<0.05), and there was significant differences in the apoptosis efficiency between pSuppressor Neo vector and the MDR1siRNA or MDR3 siRNA vector (P<0.01); 2) The reversal effect of MDR1 siRNA is higher than that of MDR3 siRNA (P<0.05); 3) The expression of MDRI and MDR3 mRNA can be restrained by pSuppressor Neo MDR1 and MDR3 siRNA respectively, and the reduction in the mRNA level was in a time-dependent manner (P<0.01). MDR1 and MDR3 gene silencing can enhance intracellular adriamycin accumulation in MCF-7/ADR cells, improve sensitivity of MCF-7/ADR cells to adriamycin, and induce cell apoptosis. The reversal effect of adriamycin resistance by siRNA of MDR1 was more effective than that of MDR3.

To explore the role and possible mechanism of apoptosis and caspase-3 activity in the development of multicellular drug resistance of ovary cancer. Ovarian cancer cell A2780 multicellular spheroids (MCS) were obtained from three-dimensional culture. Drug sensitivity of monolayer cells (MC) and MCS were respectively tested by MTT staining and cytometry. The apoptosis of MC and MCS were determined by the flow cytometry (FCM). The expression of bcl-2 and caspase-3 in A2780/MC and A2780/MCS were detected by using Western blot and caspase-3 assay kit. A2780/MC was compacted into mass after 2 days in three-dimensional cell culture model, and MCS had more than two layers of cells growing within 5 days. Compared with A2780/MC, A2780/MCS were more resistant to the anticancer drug, and the apoptosis rate was significantly lower than those of A2780/MC. The activity of caspase-3 in A2780/MCS was significantly lower than the A2780/MC. But the expression of bcl-2 in A2780/MCS was significantly higher than that in A2780/MC. It was suggested that the drug resistance of MCS might be associated with the overexpression of anti-apoptosis protein bcl-2 and the down-regulation of caspase-3 activity.

Diagnosis, Differential ; Gastric Mucosa ; pathology ; Gastritis, Hypertrophic ; pathology ; surgery ; Humans ; Male ; Middle Aged

Objective:To observe the artifacts produced by dental metal crown made from different materials on magnetic resonance imaging (MRI).Methods:Mental crows made from Co-Gr,Ni-Gr,low-Ti alloy and Pure-Ti were respectively fabricated for upper right second premolar of a ripe crossbreed dog.The 4 different casting crowns were respectively put on experimental tooth and head MRI scan was carried out with 4 sequences of 1.5 T and 6 sequences of 3.0 T MRI respectively.The existence and extent of the arti-facts were assessed and compared.Results:On both 1.5 T and 3.0 T MRI,the Go-Gr alloy crown exhibited significantly wider arti-facts than the others (ANAVO analysis,P<0.01).There were no significant differences among Ni-Gr,low-Ti alloy and Pure-Ti on 1.5 T MRI(P>0.05).On 3.0 T MRI the artifacts produced by Ni-Gr was larger than that by low-Ti alloy or Pure-Ti(P<0.05,be-tween low-Ti alloy and pure-Ti,P>0.05).By any of the test sequence either on 1.5 T or on 3.0 T,the Co-Gr alloy produced arti-facts in the most slices of MRI imaging and Pure-Ti in the least.Conclusion:Go-Gr alloy may produce larger artifacts than Ni-Gr, low-Ti alloy and Pure-Ti on MRI.

Objective To investigate the approach and effect of PDT and TTT therapeutic alliance for isolated choroidal angioma according to the lication. Methods After mydriasis,through 90D retinal mirror examination ,fluorescein fundus angiography (FFA) ,Indocyanine green angiography (ICGA) and ultrasound,the diagnosis of angiography choroidsal hemangioma was made. Position:peripapillary , six cases( tumors locating only in macula fovea were not included in this statistical area). OCT suggested that epithetlial tumors from the surface of nerve macular were affected. Period of the disease was between one week and two months . Preoperative best corrected vision acuity range was 1. 05 ～ 0.4. By ultrasonography, tumors thickenss was from 2.2mm to 4mm, and diameter was from 6.6mm to 11.2mm. OCT indicated that the height of neural epithetlium breaking away from macular area was between 156mm and 486mm. inferior retinas separated two months after morbidity. TTT was partly applied to cure tumor body ortside macdular area with IRIS Medical OculightSLX gallium arsenicde laser(810nm). Energy:per light spot shining diameter was 1 mm, and the power was between 150mw to 250mW, and time was 60s. Therapeutic standard was retinal surface changing to pale. Choroids shrunk back and retinal ptgment epithetlium prolixferated in TTT therapy area;recheck 2 weeks later. If FFA showed that active macula fovea pathological changes were not cured, PDT was performed as following :689nm laser was used , and veteporfin ( comm. ercial name : Visudyne) was injected by intravenous at a speed of 6mg/mm~2 , and 15 minutes later, the laser was irradiated for a period of 83 ～ 126s. For tumors with thicdness under 3mm,one light spot was irradiated for 83s;for rumors with thicdness above 3mm, acouple light apot was irradiated for 125s. A small quantity of hyperpigmintation occurred in the limbus,luteus retinae at the second week of follow up. All the cases were followed up for 3 months to 12 months. Results The final vision reached 0.1 ～ 1.0. OCT showed that the earliest complete recovery of limbus luteus retinae neural epithetlium was at the third week, and for another 3 cases,the recovery was at the 4th week. One case with detachment of retina partial recovered 3 months after treatment. OCT suggested that most of the macular detachment and epithetlial cystic degenerateion of neurons recovered to normal. type - B ultrasound was performed in the first three months of treatment. The tumors were difficult to be detected in two cases, and four cases had a thickness of 0. 8 ～2.2mm. At the 6th month,the detection of tumors for the six cases showed normal results. PDT was used only once in the included six cases. Conclusion PDT and TTT therapeutic alliance for isolated choroidsal angioma is safe and effective at special position of eye ground. Each treatment showed its unlque adventages in this combination with a reduction of frequentcy and cost of expensive cost of PDT, and satisfying results of the shrink of the rumors and improvement or preservation of vision.

Objective To evaluate the precision of BREEZE~(TM) glucose monitor and correlation of glucose measurements between fingertip capillary whole blood glucose(CBG)using BREEZE~(TM) glucose monitor and venous plasma glucose(VPG)using autoanalyzer.Methods All samples of venous plasma and fingertip blood from 188 diabetes or non-diabetes patients were detected for glucose level at fasting,30,60 and 120 min postprandial.BREEZE~(TM) glucose monitor and autoanalyzer measured CBG and VPG respectively.Intra-and inter- coefficients of variations were determined using 10 BREEZE~(TM) glucose monitors with normal,slight high and high glucose levels for the three lots of strips.Results The correlation coefficients between CBG and VPG were all higher than 0.950 at fasting and different postprandial time.98.94% of all measurements were in the A zone when using error-grid analysis.The relevant differences between CBG and VPG were less than 5% at different blood glucose concentrations.The intra-and inter-coefficients of variations of blood glucose values at different blood glucose concentrations using different lots of strips were within 5%.Conclusion BREEZE~(TM) glucose monitor provides high accurate and precise glucose readings on fasting and different postprandial time points over a variety of blood glucose concentrations.

Objective To measure the setup errors of patients with esophageal carcinoma during the treatment of three dimensional conformal radiotherapy (3DCRT), and to analyze the impact of setup errors on dose distribution of GTV,CTV and normal tissues around. Methods Forty-two patients with esophageal cancer treated by 3DCRT were included. The setup errors of each patient were measured once a week for 6 times by electronic portal imaging device (EPID). The setup errors were integrated into the treatment plan-ning system by moving the isocenter. Then the dose distribution of GTV, CTV and normal tissues were recal-culated. Results The systematic setup errors of the 42 patients were - 2.31 mm, - 0.55 mm and - 0.16 mm, and the random errors were 4.42 mm, 4.35 mm and 4.48 mm in the directions of lef-fight, anterior-posterior,and superior-inferior, respectively. The dose covered 95% GTV( D95 ) was reduced by 32 cGy and by 88 cGy for CTV D95. The lung V20 in the original plan and the integrated plan was 22.49% and 22.02%, respectively. The average dose of the heart in the two plans was 2077.62 cGy and 2036.23 cGy, respectively. In the original plan, no patient had maximum dose of spinal cord over 4500 cGy; While in the intergrated plan there were 18 patients had the spinal cord dose more than 4500 cGy, with a maximum dose of 5503.90 cGy. Conclusions The setup errors cause significant dose reduction of GTV and CTV, but not of the lung and heart . The maximum dose of the spinal cord may exceed 4500 cGy due to the setup errors.

The clinical analgesic effect of electro-acupuncture (EA) stimulation (EAS) on breakthrough pain induced by remifentanil in patients undergoing radical thoracic esophagectomy, and the mechanisms were assessed. Sixty patients (ASAIII) scheduled for elective radical esophagectomy were randomized into three groups: group A (control) receiving a general anesthesia only; group B (sham) given EA needles at PC4 (Ximen) and PC6 (Neiguan) but no stimulation; and group C (EAS) electrically given EAS of the ipsilateral PC4 and PC6 throughout the surgery. The EAS consisting of a disperse-dense wave with a low frequency of 2 Hz and a high frequency of 20 Hz, was performed 30 min prior to induction of general anesthesia and continued through the surgery. At the emergence, sufentanil infusion was given for postoperative analgesia with loading dose of 7.5 μg, followed by a continuous infusion of 2.25 μg/h. The patient self-administration of sufentanil was 0.75 μg with a lockout of 15 min as needed. Additional breakthrough pain was treated with dezocine (5 mg) intravenously at the patient's request. Blood samples were collected before (T1), 2 h (T2), 24 h (T3), and 48 h (T4) after operation to measure the plasma β-EP, PGE2, and 5-HT. The operative time, the total dose of sufentanil and the dose of self-administration, and the rescue doses of dezocine were recorded. Visual Analogue Scale (VAS) scores at 2, 12, 24 and 48 h postoperatively and the incidence of apnea and severe hypotension were recorded. The results showed that the gender, age, weight, operative time and remifentanil consumption were comparable among 3 groups. Patients in EAS group had the lowest VAS scores postoperatively among the three groups (P<0.05). The total dose of sufentanil was 115±6.0 μg in EAS group, significantly lower than that in control (134.3±5.9 μg) and sham (133.5±7.0 μg) groups. Similarly, the rescue dose of dezocine was the least in EAS group (P<0.05) among the three groups. Plasma β-EP levels in EAS group at T3 (176.90±45.73) and T4 (162.96±35.00 pg/mL) were significantly higher than those in control (132.33±36.75 and 128.79±41.24 pg/mL) and sham (136.56±45.80 and 129.85±36.14 pg/mL) groups, P<0.05 for all. EAS could decrease the release of PGE2. Plasma PGE2 levels in EAS group at T2 and T3 (41±5 and 40±5 pg/mL respectively) were significantly lower than those in control (64±5 and 62±7 pg/mL) and sham (66±6 and 62±6 pg/mL) groups. Plasma 5-HT levels in EAS group at T2 (133.66±40.85) and T3 (154.66±52.49 ng/mL) were significantly lower than those in control (168.33±56.94 and 225.28±82.03) and sham (164.54±47.53 and 217.74±76.45 ng/mL) groups. For intra-group comparison, plasma 5-HT and PGE2 levels in control and sham groups at T2 and T3, and β-EP in EAS group at T3 and T4 were significantly higher than those at T1 (P<0.05); PGE2 and 5-HT levels in EAS group showed no significant difference among the different time points (P>0.05). No apnea or severe hypotension was observed in any group. It was concluded that intraoperative ipsilateral EAS at PC4 and PC6 provides effective postoperative analgesia for patients undergoing radical esophagectomy with remifentanil anesthesia and significantly decrease requirement for parental narcotics. The underlying mechanism may be related to stimulation of the release of endogenous β-EP and inhibition of inflammatory mediators (5-HT and PGE2).

Heterokaryon incompatibility is a widespread phenomenon among fungi,controlled by specific loci termed het (for heterokaryon incompatibility).This review focuses on recent developments in our understanding of the molecular mechanisms of nonself recognition and the relationship between the death progresses of heterokaryon incompatibility and associated proteins in fungi.The deep research of heterokaryon incompatibility mechanism will hopefully reveal underlying principles of the evolution of nonself recognition systems and will find some effective method for settling the instability of protoplast fusant of fungi.

The study compared the traditional medical microbiology experimental teaching with a new experimental teaching pattern,with the students majoring in anaesthesia as the research object. The new pattern mainly deals with the cultivation of the students' creativity by reforming and exploring the plan,the content,the method of experimental teaching and the ways of checking the students' work, adding general and designing experiment,and working out the PPT of the experiments.The result shows the new experimental teaching pattern contributes to the cultivation of the students'abilities of performing experiment, the ways of thinking, creativity and comprehensive analysis. It's better than the traditional experimental teaching pattern.A new medical microbiology experimental teaching systerm which is suit to the students majoring in anaesthesia has been established.