Objective To explore the functional expression and temporality of MDR1 gene in bone marrow of rabbits after autologous bone marrow transplantation with MDR1 transferred bone marrow mononuclear cells. Methods The supernatant of the amphotropic virus producer cell line PA317-HaMDR1/A was collected and concentrated to cocultivate with the bone marrow mononuclear cells of the rabbits. After large dose of chemotherapy with cyclophosphamide,the transferred cells were autotransplanted into the bone marrow. The integration,transfection rate and physiological function of MDR1 gene were tested by PCR,SP immunohistochemical method and daunorubicin (DNR) extrusion test respectively. Results After autologous bone marrow transplantation had been executed for 1-4 months,the integration of MDR1 gene in genome of bone marrow mononuclear cells was detected by PCR,and the expression rates of P-gp in cells tested by SP immunohistochemical method were 9.5%,8.5%,6.0% and 3.5% respectively. The physiological function of MDR1 gene in bone marrow cells was proved by DNR extrusion test. Conclusion After the autotransplantation with bone marrow mononuclear cells transferred by MDR1 gene,the MDR1 gene can implant into the bone marrow of rabbits and has expressed functionally for 4 months,which has provided a basis for further research on chemoprotection experiment of the MDR1 gene transferred into the bone marrow cells.

Objective To evaluate the feasibility, safty and methology of transradial coronary angiography Methods Transradial coronary angiography was carried out in 304 patients Results Transradial coronary angiography and ventriculography were successful in 291 patients (95 7%) Conclusion Radial artery seems to be both feasible and safe for coronary angiography, with a very low risk of local complication and more comfort for patients

BACKGROUND: Heme oxygenase-1 (HO-1) plays an important role on preventing tissues and organs from oxidant stress injury, which remains currently one of the most active areas of investigation. OBJECTIVE: To study HO-1 over-expression on the survival and fiver function of rats after reduced-size liver transplantation by constructing recombinant adenovims Ad5-HO-1.DESIGN, TIME AND SETTING: Randomized controlled animal study was performed in the Institute of Pediatrics, Children Hospital Affiliated to Chongqing Medical University from September 2003 to March 2005. MATERIALS: Seventy-four SD rats were used to establish in situ reduced-sized liver transplantation models. METHODS: Recombinant adenoviral vector encoding Ha-1 gene (AdS-HO-1) was generatred by molecular biology method, wluch was administered to donors via voln at 48 hours before transplantation. All rats were randomly divided into control group(n=12),saline group(n=12), cobalt protoporphyrin (CoPP)group(n=13),Ad5-HO-1 group(n=13),Ad5-green fluorescent protein(GFP)group(n=12),and Ad5-HO-1+zinc protoporphyrin (ZnPP)group (n=12).Livers of donor wefe harvested and stored for 24 hours at 4℃ in HTK solution. Before it is implantedinto recipient. MA IN OUTCOME MEASURES: Survival rate and liver function; portal vein blood flow monitored 2 hours after transplantation by Color Doppler How Imaging; pathological changes of transplanted liver tissue observed by HE staining;HO-1 activity, tumor necrosis factor-alpha(TNF-α),Bcl-2 and Bax expression detected by immunohistoehemical staining; changes of HO-1,TNF-α,bcl-2 and bax mRNA detected by molecular viewpoint. RESULTS: Survival rate in the Ad5-HO-1 group was significantly higher than that in the saline group at al,7,and 21 days after liver transplantation(P＜0.05).Glutamic pyruvic transaminase decreased in the Ad5-HO-1 group as compared to that in the saline,Ad5-GFP,and Ad5-HO-1+ZnPP groups(P＜0.05);portal venous blood flow significantly increased 2 hours after transplantation(P＜0.05);HO-1 activity also significantly increased(P＜0.05).RT-PCR and immunohistochemical staining showed that HO-1 and bcl-2 expressions increased(P＜0.05),but bax and TNF-αexpressions decreased(P＜0.05).CONCLUSION:Ad5-HO-1 significantly induces high expression of HO-1 increases portal venous blood flow within 2 hours alter liver transplantation, and promotes liver functional recovery so as to prolong survival time of rats after liver transplantion.

Objective To study the feature of neonatal infections and characteristics of antibiotic treatment in a tertiary children ' s hospital. Methods Clinical data including incidence of infection, primary disease,species of bacteria, complication and antibiotic utilization in hospitalized patients from Jan. 2010 to Dec. 2012 were retrospectively reviewed using their medical records. Results Among 1826 patients admitted to neonetal surgery ward, 542 infants ( 29. 7％) were with infection. The incidence of antibiotic resistance was 23. 51％. The top five infectious diseases were:perianal abscess, necrotizing enterocolitis, colicitis, omphalitis and subcutaneous gangrene. 12 cases of multi-resistant infection were cured by non-restricted antibiotics. 109 were cured by restricted antibiotics. And other 7 were cured by special antibiotics. No death nor multi-resistant nosocomial infection were found. Risk factors including multi-site infection, premature or low birth weight infants, liver, kidney or heart dysfunction,fever lasting more than 3 days after antibiotic therapy, septic shock, sepsis, digestive tract perforation and peritonitis,were vital in choosing specific antibiotics. Conclusions Infection is one of the most common diseases in neonatal surgery ward, with major pathogens sensitive to antibiotics. The clinical characteristics and drug sensitive test are conductive to the reasonable use of antibiotics. Special antibiotics can be used directly in patients with risk factors Clinical doses of antibiotics in neonates depend on the monitoring of drug concentration.

Objective To investigate the effect of stanozolol on nitrogen balance ,grip strength and clinical outcomes of criti‐cal patients with high nutrition risk .Methods We enrolled patients who were admitted to the ICU of Guizhou provincial Hospital during the time period from January 2014 to June 2014 and ,as patients with high nutrition risk .Patients ,who received same base nutritional support program ,were divided into two groups .Treatment group who were treated with stanozolol administrated with gastric or jejunal tube for 7 days by 4 mg Tid .The control group whose members underwent placebo simultaneously with the treat‐ment group .The nitrogen balance ,grip strength of both groups was measured when at admitted and 4th as well as 7th day .Prealbu‐min ,total bilirubin ,alanine aminotransferase ,and aspartate aminotransferase were measured when at the same time and before leave hospital .The duration of the mechanical ventilation ,ICU stays ,hospital stays and mortality within 28 days were recorded .Results There was no statistical significance in the differences between all the indicators of the two groups at admission(P>0 .05) .The du‐ration of mechanical ventilation ,ICU stays ,hospital stays were decreased significantly in the treatment group (P<0 .05) compared to the control group .But the mortality within 28 days had no significant difference between two groups (P>0 .05) .Nitrogen bal‐ance ,prealbumin ,grip strength and liver function parameters in the treatment group were significantly higher than they were been at admitted and control group at 4th and 7th day (all P<0 .05) .Liver function parameters of treatment group gradually decreased to the normal range before discharge .Conclusion In critically ill patients treated with anabolic steroid stanozolol ,can promote protein synthesis ,reduce muscle and other lean tissue decomposition ,improve clinical symptoms ,short the length of hospital stay and ICU stay .But we should pay more attention on liver function in critically ill patients who treated with stanozolol .

Objective To explore the mechanism and safety of miltidrug-resistance gene 1(MDR1)transfection into the bone marrow mononuclear cells of rabbit in vitro with the adenovirus vector promoted by ultrasonic microbubble.MethodsBone marrow mononuclear cells of rabbits were collected and divided into 5 groups after cultured in the 6-well plate according to the different experimental conditions(MDR1 gene was transferred into the cells with or without ultrasound irradiation and microbubbles)：conventional culture group (A)，Ad5-MDR1 group(B)，Ad5-MDR1+ultrasound irradiation group(C)，Ad5-MDR1+microbubbles group(D)，Ad5-MDR1+ultrasound irradiation+microbubbles group(E).The positive transfection rate of MDR1 gene in mononuclear cells of different groups were tested by flow cytometry,and the survival rate of cells in different periods were tested by trypan blue exclusion method.Moreover,the appearance and ultramicrostructure of cells were observed by electronmicroscope.Results①The transfection rate of MDR1 gene in different groups were 0.39%±0.11%，5.03%±0.35%，4.93%±0.38%，5.25%±0.80%and 19.93%±1.51%respectively.The transfection rate of MDR1 gene in group E was higher than those in other groups(P＜0.05).②Compared with those in control groups(group A，B，C and D)，the transfection rate in group E was significant raised by ultrasound irradiation and microbubbles.However,there were no significant difference in survival rate of cells between the five groups(P＞0.05).③After ultrasonic irradiation，there were transient holes on the cell membrane，which could disappear after irradiation by ultrasound for 24 hours.And the temporary swelling of organelles was reversible.Conclusions Microbubbles irradiated by ultrasound can cause small transient holes on eell membrane and increase permeability of it，and enhance the transfection of MDR1 gene in bone marrow mononuclear cells with the adenovirus vector，which is safe and available.

Objective To analyze the pattern of antibiotic use and antibiotic resistance tendency of gastrointestinal surgery in a tertiary children′s hospital .Methods 2 625 patients(which account for 27 .52% of all the hospilitalized patients ,the resistant rate was 15 .70% ) detailed morbidity ,entity ,bacteria ,complication ,antibiotic utilization was retrospectively reviewed using the hospital medical records from 2010 to 2012 .Results 2 625 patients the percentages of the top five disease category were :appendicitis ac-counting for 40 .72% ,perianal abscess accounting for 21 .53% ,periappendiceal abscess accounting for 9 .30% ,necrotizing enterocol-itis accounting for 3 .73% ,omphalitis accounting for 2 .93% .The top three pathogen were :escherichia coli ,klebsiella pneumoniae subsp ,staphylococcus aureus respectively .255 multi-resistant bacteria of the superficial infection patients and 157 of the invasive in-fection patients .49 multi-resistant infections were cured by first or second generation of cephalosporins and penicillinase-fast peni-cillin ,and 346 were cured by third or forth generation of cephalosporins and penicillinase-fast penicillin ,and 17 were cured by car-bapenem or vancomycin .No dead or multi-resistant hospital infectious case was reviewed .Conclusion The sensitive rates of surgi-cal infected patient were 84 .3% ,and opportunistic pathogen infection was the main characteristics .To aware the clinical character-istics and drug sensitive test is conductive to the reasonable use of antibiotics of severe infections .The cases of superficial resistant infection or invasive non-resistant infection tend to use restricted antibiotics .The cases of invasive resistant infection tend to use special antibiotics .

Objective To explore the effect of ranolazine on the fast sodium channel current (INa) in rabbit atrial myocytes and the existence of use-dependent blockade. Methods Standard whole cell patch clamp technique was used to study the effect of ranolazine on the fast sodium channel current and the use-dependent blockade caused with different frequencies (1Hz, 3.3Hz and 5Hz) to stimulate the cells. Results The 30μmol/L ranolazine significantly reduced INa with an IC_(50) value of (25.6±1.8)μmmol/L and produced a frequency-dependent inhibitory effect on INa with obvious use-dependence. Conclusion Ranolazine can inhibit the fast sodium channel current in rabbit atrial myocytes and indeed has a use-dependent effect.

Objective:To observe the killing effect of irradiated peripheral blood mononuclear cells (PBMCs) at low dose combined with apogossypolone (ApoG2) on cultured human prostate cancer PC-3 cells.Methods:Human PBMCs were irradated by gamma ray at 1 gray,the irradiation dose rate was 17 Gy/min.The experiment were divided into PC-3 tumor cell control group,PC-3 cells with irradiated and non-irradiated PBMCs co-culture groups,ApoG2 treatment group,irradiated PBMCs and ApoG2 co-treatment group.Acridine orange/ethidium bromide (AO/EB) staining and MTT method were used to observe the killing effect of PBMCs and/or ApoG2.Results:The killing activity of irradiated PBMCs group and ApoG2 treatment group were obviously increased and were higlaer than that of non-irradiated group (P＜0.05).The killing activity of combined group were much higher than that of irradiated group and ApoG2 treatment group (P ＜0.01 ).Conclusion:Irradiated PBMCs at low dose combined with ApoG2 can enhances the anti-tumor effects markedly.

Objective To establish a method for the isolation and identification of platelets.Methods 10 healthy volunteers were selected to collect the EDTA anticoagulant venous blood of 3 tubes,each tube was 2 ml,which was divided into the whole blood cell tube,platelet rich plasma (control group),and stepped centrifugal platelet extract (experiment group).Platelet was isolated by simple centrifugation method(PRP) and stepped centrifugal method.The two groups were full blood count and analyzed by microscopic morphology and platelet activity test.Leukocyte specific HGB gene and platelet mitochondrial ND1 gene content was analyzed by real time PCR.Results Platelets were extracted and detected in control group and experimental group.Platelets were found and white blood cells and red blood cells were not remained in experimental group.Platelets and sporadic white blood cells were found in control group.The platelet pick up rate of experiment group was significantly higher than control group,the difference was statistically significant.Experimental gene content HGB of experiment group was significantly lower than control group,the difference was statistically significant (t=-3.281,-2.865,P＜0.05).ND1 gene content of experiment group higher than the control group,the difference was not statistically significant.There was no significant difference for platelet activity test between experimental group and control group (t=-0.046,-0.799,P＞ 0.05).Conclusion A isolation and identification method of stepped centrifugal platelet was established.The method can be used for the study of platelet gene and the functional analysis of platelets.