OBJECTIVE Comparing and analyzing the use of antibacterial drugs in our hospital for nearly 10 years,to guide more standardized and more rational use of antibiotics in clinical pediatrics.METHODS The use of antimicrobial drugs among 3711 pediatric cases in our hospital from 1999 to 2008 was comparatively analyzed.RESULTS The usage rate of antibacterial drugs among them in our hospital for nearly 10 years was 93.45%.The use of top six kinds of antimicrobial drugs constantly changed every year,so did the use rate.The rate of combined application was more than 43%,except for 17.67% in 2004,and the kinds of combined application were increasing year by year.The usage rate of second-line antibiotics was increasing year by year,increased from 7.25% in 2000 up to 71.89% in 2008.The preventive drug use was reduced year by year.CONCLUSIONS Using the antimicrobial agents in pediatric is more reasonable than the other departments in our hospital.However,there are some unreasonable phenomena.Antimicrobial agents use should be strengthened in the field of guidance,supervision and management.

BACKGROUND: As a method to effectively improve organic tolerance and velocity, altitude training has been gradually accepted by the coaches,and it is a problem to be solved to find the optimal altitude.OBJECTIVE: To establish rat models of stimulating altitude training, and observe the changes of myocardial cell structure in the altitude training, so as to provide scientific animal models for the physiological studies of altitude training, and provide more scientific theoretical evidence for the altitude training of athletes.DESIGN: Based on the division of altitude region in aerospace medicine,the experimental animals were divided into different altitude training groups and control groups. Morphological characters of cellular and subcellular structures were observed under electron microscope, the biochemical analysis of cardiac troponin T were performed, and then the myocardial injury was completely analyzed from two pathways of morphology and biochemistry.SETTING: Laboratory of Sports and Human Sciences, Tsinghua University.MATERIALS: The experiments were carried out in the Laboratory of Sports and Human Sciences, Tsinghua University from January 2000 to December 2002. Totally 372 male SD rats were used. The experimental materials included hypobaric oxygen chamber, Japanese H-500 transmission electron microscope and the kit for testing serum cardiac troponin T index.METHODS: After adaptive swimming training for 1 week, all the 372 rats were randomly divided into 9 groups and 54 subgroups, only 6 groups of them were analyzed in this experiment, including stimulating acute altitude hypoxia training at 4 000 m group (n=32), hypoxia without training group (n=24), stimulating acute altitude training at 4 000 m followed by reoxygen training for 1 week group (n=32), stimulating acute altitude training at 4000 m followed by reoxygen training for 2 weeks group (n=32), increasing altitude training group (n=32) and plain training control group (n=24). Rats in the plain training control group lived and trained in normal conditions,and those in the other groups all trained with swimming in the stimulating altitude environment for 4 weeks. They were trained for 7 days every week,once a day and 60 minutes for each time for 4 weeks. The serum cardic troponin T index of rats was detected and observed with enzyme-linked immunoabsorbent assay (ELISA). After stimulating acute altitude training at 4 000 m for 4 weeks followed by reoxygen training for 2 weeks, the rats were killed, and a small piece of myocardial tissue was taken from left and right ventricles, then double-stained with uranyl acetate and lead citrate,and the ultrastructure of the sections was observed with Japanese H-500 transmission electron microscope.MAIN OUTCOME MEASURES: The serum cardiac troponin T index of rats trained at different altitude, and the results of spectrum and observation of myocardial ultrastucture under electron microscope were mainly observed.RESULTS: ① Changes of body mass: The myocardial increasing rate in the hypoxia training group was significantly different from those in the hypoxia without training control group and plain training control group (P ＜ 0.01), and there was also significant difference between the latter two groups (P ＜ 0.05). The increasing rate of body mass was significantly different among the hypoxia training group, hypoxia without training control group and plain training control group (22.41％, 28.14％, 33.48％,P ＜ 0.01-0.05). ② Results of cardiac troponin T: Compared with the hypoxia without training group, the value of cardiac troponin T in the hypoxia training groups was significantly different, and significantly increased with weeks of training (P ＜ 0.01). During the hypoxia altitude training below 4 000 m for 1-4 weeks, the value of cardiac troponin T ranged in creased from 0.09 to 0.128 μg/L, and it gradually recovered to 0.09 μg/L after reoxygen training in plain for 2 weeks. ③ The results of observation under electron microscope showed that after acute hypoxia training at 4 000 m for 4 weeks, myocardial mitochondrial structure occurred mild swelling with the prolongation of training, and for the rats received reoxygen training in plain for 2 weeks, the thick and thin myofilaments of myocardial fibers formed light and dark bands were clear and distinct, and the myotomes arranged orderly, besides clustered or scattered glycogen granules, a great amount of mitochondria also existed among muscular bundles, no swelling, degeneration of vacuoles, lysis necrosis, interstitial edema and other pathological changes were observed.CONCLUSION: It is safe to train rats at any altitude below 4 000 m. The acute exercise training of great intensity directly at 4 000 m will cause the mild injury of cardiac subcellular structure, and the increase of cardiac troponin T concentration. The training by means of increasing altitude has slighter influence on the myocardial injury, and the recovery after training is also faster.

High-speed counter-current chromatographic ( HSCCC ) method was successfully used to separate and purify prim-O-glucosylcimifugin and 5-0-methylvisammioside from Saposhnikovia divaricata(Thicz. ) Schis-chk. ,a traditional Chinese herb,with a solvent system composed of ethyl acetate-n-butanol-water(2:7:9,V/V). The lower phase of the system was used as the mobile phase at the flow rate of 2.0 mL/min,and the upper phase was used as the stationary phase. The separation produced a total 13.9 mg prim-O-glucosylcimifugin and 25.0 mg 5-0-methylvisammioside with the purity of 98. 1% and 99.2% determined by high performance liquid chromatography (HPLC) from 316 mg of crude sample from S. divaricata. The structures of isolated compounds were identified by ESI-MS,~1H NMR and ~(13)C NMR.

AIM:To investigate the effect of 18 alpha-glycyrrhetinic acid (18α-GA) on delaying the senescent progress and promoting the proliferation in late-passage bone marrow mesenchymal stem cells ( BMSCs ) .METHODS:Late-passage BMSCs were incubated with 2.0 mg /L 18α-GA or the same volume of DMSO for 30 d, and the cells were harvested to determine the proteasome activity .The expression of senescence-related proteins p53, p21 and p16 was detec-ted by senescence-associated β-galactosidase ( SA-β-Gal) staining and Western blot .The cell proliferation , the expression level of cell cycle-related proteins and cell cycle distribution of the cells were measured by CCK -8 assay, BrdU incorpora-tion, Western blot and flow cytometry.RESULTS:Compared with DMSO group, the proteasome activity in 18α-GA group increased significantly by about 0.2 times (P<0.01).SA-β-Gal-positive cells in 18α-GA group decreased, and cell stai-ning was lighter.The contents of p53 and p21 in 18α-GA group were decreased (P<0.05).The results of CCK-8 assay showed that the A value in 18α-GA group was 0.3 times higher than that in DMSO group (P<0.01).BrdU incorporation showed the increased proliferation in 18α-GA group compared with DMSO group ( P<0.05).The cells in G1 phase in 18α-GA group decreased significantly compared with DMSO group , while the cells in S phase increased significantly ( P<0.05).The expression level of cyclin D1 in 18α-GA group was 2.8 times higher than that in DMSO group (P<0.01), and the CDK4 level was 1.4 times higher than that in DMSO group (P<0.05).CONCLUSION:Activation of the pro-teasome activity by 18α-GA delays the aging process in the BMSCs and promotes the cell proliferation via up -regulation of the cell cycle-related proteins .

Objective To value the effect and safety of butyl phthalide sequential treatment in cerebral infarction of branch sclerosis of arterial congee appearance. Methods Sixty patients with acute cerebral infarction were randomly divided into treated group( n = 31)and control group( n = 29). According to the condition of illness,all patients were given aspirin,atorvastatin calcium,and the injection of ozagrel sodium intravenous;controlled the blood pressure,blood sugar,blood lipid,and treated complications posstively;take the early rehabilitation of nerve treatment afte the illness was in stable condition. Butyl phthalide was used in the patients of treated group(100 ml,twice per day,intravenous drip,during 14 days period therapy,and then 0. 2 g oral,third per day),besides the routine therapy. The degree of neural function defect score( NIHSS)and activities of daily living score(BI)between two groups were observed before and after treatment. Corresponding adverse consequences were recorded. Results Compared with pretreatment,the NIHSS of postreatment at the 14th day in treat and control groups were decreased(treated group:(4. 36 ± 3. 11)vs.(11. 42 ± 3. 20);control group:(6. 12 ± 2. 67)vs.(11. 64 ± 3. 43),P < 0. 05,and the treated group was significantly lower than control group(F inner groups = 2. 125,P < 0. 01;F between groups = 18. 63,P < 0. 01;F cross groups = 25. 34,P< 0. 01;P < 0. 05). The BI of postreatment in two group were increased(treated group:(86. 72 ± 8. 44)vs. (26. 54 ± 13. 36);control group:(75. 96 ± 9. 86)vs.(26. 38 ± 13. 02)),and the treated group was significantly lower than control group(F inner groups = 29. 27,P < 0. 01;F between groups = 32. 48,P < 0. 01;F cross groups= 42. 41,P < 0. 01;P < 0. 05). There was no the adverse reactions. Conclusion Butyl phthalide sequential treatment can improve the NIHSS and BI of cerebral infarction of branch sclerosis of arterial congee appearance and have a better therapy effect.

BACKGROUND:Cationic liposome-mediated celltransfection is reliable and repeatable. However the transfection efficiency is often low.
OBJECTIVE:To study the optimized methods for gene transfection mediated by liposome into N2a cells (mouse neuroblastma cells).
METHODS:Using traditional adherent method and improved suspension method, 500 ng recombinant plasmid pcDNA3-GFP carrying green fluorescence protein was transfected into N2a cells in 24-wel culture plate, which was mediated by 1.5μL Lipofectamine?LTX Reagent. The expression of green fluorescent protein was observed by inverted fluorescence microscope, and the transfection efficiencies at different transfection ways were calculated. By using improved suspension transfection method, 500 ng plasmid DNA was transfected with different doses of Lipofectamine?LTX Reagent (1.0, 1.5, 2.0, 2.5μL). The optimal ratio of liposome and DNA was explored.
RESULTS AND CONCLUSION:The transfection efficiency of suspension transfection method was significantly higher than that of the tranditional adherent method (P<0.01) when using 1.5μL liposome/500 ng DNA. The transfection efficiency of the 1.0, 1.5, 2.0, 2.5μL Lipofectamine?LTX on 500 ng plasmid DNA was respectively (76.60±3.85)%, (80.00±4.17)%, (88.00±5.89)%, (54.96±4.23)%. It showed the 500 ng DNA and 2.0μL liposome achieve the highest transfection efficiency.

Objective To evaluate the effect of external direct current shock on left atrial(LA) diamension and volumes after cardioversion for atrial fibrillation(AF) and the relation between LA size and function.Methods The initial study population included 68 patients with AF.According to cardioversion patterns, the patients were divided randomly into cardioversion with direct-current shock(36 patients) and cardioversion with drugs(32 patients). Echocardiographic evaluations included LA size and volume,LA passive and active emptying volumes were calculated,and LA function was measured as LA ejection force (LAEF).The correlation between LA diameters and volumes and LA systolic function was analyzed. Results LA was dilated in all the patients during AF and the superior-infra dimensions of LA decreased after restoration of sinus rhythm in the patients reverted with external direct current shock and the patients reverted with durgs (P

BACKGROUND:It is not fuly understood that whetherp53 inhibitor can directly intervene in the viability of bone marrow mesenchymal stem cels and the possible mechanism. OBJECTIVE:To investigate the effect of p53 inhibitor, PFT-α, on the aging process of bone marrow mesenchymal stem cels in late-phase amplification and to discover the key target to delay the replicative senescence of human bone marrow mesenchymal stem cels. METHODS:The expression levels ofp53,p21, andp15 mRNA in human bone marrow mesenchymal stem cels in both early and late-phase amplification were detected by quantitative PCR assay. Then, human bone marrow mesenchymal stem cels in late-phase amplification were respectively treated with 20 μmol/L PFT-α or an equivalent amount of dimethyl sulfoxide for 2 weeks. The positive rate of aging cels was determined by SA-β-Gal staining. The apoptosis was detected by TUNEL staining. Human bone marrow mesenchymal stem cels were treated with 300 μmol/L H2O2 for 30 minutes, and then celular anti-oxidative stress capacity was detected by cel counting kit-8 assay. RESULTS AND CONCLUSION:The quantitative PCR assay showed that the mRNA expression level ofp15, p21 andp53 in human bone marrow mesenchymal stem cels in late-phase amplification was significantly increased (1.45±0.23), (1.51±0.14) and (1.78±0.14) times as much as that in early phase amplification (P < 0.05). The positive rate of aging cels in PFT-α group was significantly lower than that in the dimethyl sulfoxide group[(41±5)%vs. (63±7)%,P < 0.05)]. However, there was no significant difference in apoptosis rate between PFT-α group and dimethyl sulfoxide group. After treatment with H2O2, the absorbance value in the PFT-α group was(1.27±0.13) times as much as that in the dimethyl sulfoxide group (P < 0.001). The above results demonstrate that the activation ofp53 signaling pathway may be an important factor of causing aging of human bone marrow mesenchymal stem cels. Application ofp53 inhibitor PFT-αcan enhance the anti-oxidative stress capacity of human bone marrow mesenchymal stem cels in late phrase amplification.

BACKGROUND: BALB/c and C57BL/6 mice are two inbred strains, but after voluntary movement, there is no report on how to scientifical y reasonably select behavioral experiment methods and indicators and to evaluate the learning and memory abilities of mice.
OBJECTIVE: To analyze and compare the behavioral indicators between BALB/c and C57BL/6 mice fol owing voluntary wheel running, to explore the effect of exercise on learning and memory, and to provide a reference for selecting reasonable behavioral indicators.
METHODS: 2.5-month-old BALB/c and C57BL/6 mice were randomly divided into control and voluntary wheel running groups. Independent running wheel movement of mice was recorded with VitalView system. 4 weeks later, newborn neurons were labeled via DCX immunofluorescence. Spatial learning, memory and exploration abilities were compared through new arm test, new object recognition test and Morris water maze test.
RESULTS AND CONCLUSION: (1) The mean spontaneous activity of BALB/c mice daily was 2.56 fold of that of C57BL/6 mice during wheel running (P < 0.001). (2) Hippocampal DCX-positive cel s in exercised BALB/c and C57BL/6 mice were more than those in control group. (3) Meantime, exhibited by higher frequencies to explore new arm or object, and longer time and distance of moving around them, the learning and exploring capability was improved after exercising (P < 0.001), especial y in BALB/c mice. (4) Wheel running C57BL/6 mice exhibited progressed spatial learning and memory abilities compared with control mice in Morris water maze test, characterized by decreased latency to target, elevated target crossings and longer time or distance in quadrant zone (P < 0.05). However, there was no significant difference between wheel running and control BALB/c mice. Taken these data together, voluntary wheel running contributed to hippocampal neurogenesis of BALB/c and C57BL/6 mice, accompanied by the change of learning and memory capability, which could be detected properly via both new arm test and new object recognition test, but for Morris water maze test, C57BL/6 mice might be superior to BALB/c mice.

Objective To investigate application of diffusion tensor tractography (DTT) on evaluating motor dysfunction degree of patients with cerebral infarction and explore the relationship between infracted focus and cortical spinal tract(CST).Methods Forty patients with cerebral infarctions in the different periods and 30 healthy volunteers were enrolled our study.They all underwent DTT check and then established CST.Meantime,all the patients were measured ADL score according to activities of daily living (ADL) on admission and two months after the onset.Results DTT image was made for both in patients and health control people.The CST was built in people in control group and its shape was like the adults'.Its shape was from the precentral gyrus down to internal capsule and continued to pontine and medulla oblongata.Each successive was form good consistency.The DTT image of patients' health side was like normal and injury corticospinal tract was affected by influence of infarction area at different level,manifested as loss of discontinuity and anatomical configuration consistency.Based on the degree of CST,there were 9 cases in grade Ⅰ,20 cases in grade Ⅱ and 11 cases at grade Ⅲ.DTT image showed that the damage level of CST had an negative relation with ADL score at 2 month after therapy (r =-0.914,P ＜ 0.05).Conclusion DTT can show the spatial positions of infarctions and CST at non-invasive lesion and analyze the influence of infarctions and corticospinal tract.Therefore,it can serve as the objective proof for judging the injury of the motor function and evaluating prognosis.