Objective The construction of the reconstructed acellular extracellular matrix(REAECM) of bone and preliminary investigation of biocompatibility of REAECM was performed to supply the experimental basis for the extracellular scaffold of bone tissue engineering. Methods Applying the technique of culture in vitro, biocompatibility of the REAECM cultured with osteoblasts was observed using phase-contrast microscope and scanning eletron microscope at 1-4 weeks. Results The osteoblasts could attach and proliferate, even grow well into the foam of REAECM.Conclusion REAECM is an excellent scaffold material for bony tissue engineering construction, which pocesses a morphological structure of three-dimensional foam. The REAECM showed a good biocompatibility with osteoblasts co-cultured in vitro. There was prospective signs of REAECM to the attachment, proliferation of the osteoblasts

Objective:To study the inhibitory effect of clarithromycin on angiogenesis induced by b-FGF. Methods:TheMatrigel implant assay was used. Matrigel (500?l) containing b-FGF and heparin were injected subcutaneously into the ab-domen of mice and were harvested 5 d later. The amount of hemoglobin and micro-vascular area present in the implant weremeasured and cotnpared. The mice were given either CAM (study group) or the same volume of glucose (vehicle group) oncea day by gastric intubation. Treatmen started 3 d before Matrigel implant and continued until the end of study. Results:Clar-ithromycin reduced hemoglobin content and micro-vascular area in Matrigel implant at high dosage(≥40 mg/(kg?d). Con-clusion:These data demonstrate that clarithromycin is a potent inhibitor of angiogenesis and may has possible therapeutic value in controlling pathologic angiogenesis.

Objective To compare the differences between two statistical methods for evaluating non-sensitivity of pathogenic bacteria to antimicrobial agents,and explore effect of non-consideration of clinical background on evalua-ting extent of bacterial resistance.Methods Data of Staphylococcus aureus and Acinetobacter spp .in a hospital in the first half year of 2008,2010 and 2013 were collected and conducted statistical analysis with two methods (me-thod 1 :based on all clinically isolated bacteria;method 2 :based on infection-related non-repetitive bacteria),two methods for evaluating bacterial non-sensitive rates to antimicrobial agents were compared.Results The non-sensi-tive rates of Acinetobacter spp .to various antimicrobial agents :statistical results by using method 1 were generally higher than those using method 2,absolute difference between two statistical methods was 10.46%-33.77%;the non-sensitive rates of Staphylococcus aureus to various antimicrobial agents :except compound sulfamethoxazole in 2010 and 2013(difference were 6.17% and 10.21 % respectively),penicillin G (difference was 3.86%),erythromy-cin (difference was 2.71 %),and azithromycin in 2013 (difference was 2.43%),statistical results by using method 1 were generally higher than those using method 2,absolute difference between two statistical methods was 0-18.04%.Conclusion There are deviation in the non-sensitive rates of bacterial strains to antimicrobial agents by using two different statistical methods,deviation is larger in Acinetobacter spp ..The resistance level might be incorrectly higher when evaluating the resistance status without considering clinical background of bacteria.

AIM: Ischemic post-conditioning ( PC) plays an important role in cardioprotection from ischemia /reperfusion ( I/R) injury in the young heart but not in the aging hearts .The physiological and pathological roles of hydrogen sulfide ( H2 S) in the regulation of cardio-vascular functions have been recognized .Whether H2 S is involved in the recovery of PC-induced cardioprotection in the aging hearts is unclear.METHOD:The male Wistar young rats (3-month-old), the aging rats (24-month-old), primary cultured cardiomyocytes and the aging cardiomyocytes induced by D-galactose suffered from I/R (or H/R) and PC.RESULTS:I/R (or H/R) decreased H2S production rate and cystathionine γ-lyase (CSE) expression, aggravated cardiomyocyte damage , apoptosis, myocardial infarct size and oxidative stress, reduced cardiac function, increased the levels of Bcl-2, cleaved caspase-3 and caspase-9 mRNA and proteins, promo-ted the release of cytochrome c and mPTP opening, down-regulated the phosphorylation of ERK 1/2, PI3K, Akt and GSK-3βand mito-chondrial membrane potential , up-regulated the phosphorylation of IκBα, NF-κB, JNK2 and STAT3, and inhibited PKC-ε transloca-tion and mitochondrial ATP-sensitive K channels (mitoKATP) in isolated young and aging hearts as well as normal and aging cardiomyo-cytes.PC suppressed myocardial I/R injury in the young heart but not in the aging hearts .Supply of NaHS not only increased PC-in-duced cardioprotection in the young hearts and cardiomyocytes , but also attenuated I/R injury and significantly recovered the cardiopro-tective role of PC in the aging hearts and cardiomyocytes .CONCLUSION:The exogenous H 2 S restores PC-induced cardioprotection through inhibiting oxidative stress via the down-regulation of NF-κB and JAK2-STAT3 pathways and mPTP opening by the up-regulation of ERK1/2-GSK-3β, PI3K-Akt-GSK-3βand PKC-ε-mitoKATP pathways in the aging hearts and cardiomyocytes .These findings provide a novel potential target for the treatment of aging ischemic cardiomyopathy .

AIM: To study the effect of calcium sensing receptor (CaSR) on icariin (ICA) induced mouse embryonic stem cells ( mESCs) to differentiate into cardiomyocytes in vitro.METHODS:mESCs were cultured to embry-oid bodies ( EBs) by direct suspension method and the differentiation of EBs into cardiomyocytes was induced by ICA.The expression of cardiac specific proteinsα-actinin and cardiac troponin-I ( cTnI) was analyzed by Western blot and immuno-fluorescence.The differentiation rate was determined by flow cytometry.The ultrastructure of the derived cardiomyocytes was further characterized by transmission electron microscopy.The expression of cardiac-specific transcription factors Nkx2.5 and GATA-4,as well as CaSR was detected by Western blot.RESULTS: After induction with ICA, the positive characteristics of myocardial cells appeared in the EBs cultured for 2 d.The expression of cardiac-specific sarcomeric pro-tein actinin (α-actinin) and cTnI showed an overall upward trend by Western blot in different phases of ICA induced differ-entiation.The expression of CaSR, Nkx2.5 and GATA-4 was the highest at an early stage of ICA-induced differentiation. Neomycin (an activator of CaSR) up-regulated CaSR, NKx2.5 and GATA-4 expression in the EBs at early stage of ICA-in-duced differentiation, all of which were reversed by NPS2390 ( an inhibitor of CaSR) .CONCLUSION:CaSR is function-ally expressed in mESC-derived cardiomyocytes, and activation of CaSR is involved in the differentiation of mESCs into car-diomyocytes by facilitating the expression of NKx2.5 and GATA-4.

Purpose　To explore the possibility and the effect of therapeutic bronchial asthma by antisense oligonucleotid.　Methods　Based on the IL-5 cDNA sequence of mouse,a segment of antisense oligonucleotid was designed and synthetized.5′-labeling of antisense oligonucleotid was signed by T4 PNK in order that the efficiency of stearylamine liposome in transfe-ting antisense oligonucleotid can be evaluated. Astham model was duplicated with ovalbumin (OVA) absorbed to aluminum hydroxide. T lymphocytes of mice were separated by nylon fiber method,then T lymphocytes transfected a different content of antisense oligonucleotid with stearylamine phys. positive liposome were cultured respectively in order to observe the effect of antisense oligonucleotid on IL-5 produced by T lymphocytes. IL-5 levels in the supernatants of T lymphocytes culture were determined by ELISA.　Results 　Stearylamine liposome could markedly increase the efficiency of antisense oligonucleotid transfection. The efficiency of antisense oligonucleotid transfection was the best at 1∶15 m/m(antisense oligonucleotid and SA liposome) and it was increased approximately 12 times.In healthy and asthma Balb/c mice, IL-5 was not detected in the supernatants of T lymphocytes culture without challenge with OVA.However,IL-5 was increased markedly in the supernatants of T lymphocytes culture challenged with OVA. After transfecting a different concentration antisense oligonucleotid, IL-5 levels in the supernatants of T lymphocytes culture were significantly lower than those in control cells without antisense oligonucleotid transfection. IL-5 levels decreased from (44.60±6.23) to (30.70±7.362),(17.20±6.181) and(8.16±2.34)pg/ml respectively. And IL-5 synthesis was inhibited by 31.17% , 61.43% and 81.7% respectively. 　Conclutions　IL-5 synthesis could be obviously inhibited by antisense oligonucleotid and showed a markedly relation between quantitative and effect. It is supported that the production of IL-5 be inhibited through preventing the transcription of IL-5 from T lymphocytes. The study provides foundation for antisense gene therapeutic asthma.

Objective To investigate the incidence of extra intestinal organ damage in infants with acute rotavirus (RV) infection,the relative risk factors in patients with extra intestinal organ damage,the significance of procalcitonin(PCT)in those infants with multiple organ injury.Methods One hundred and three infants with acute diarrhea whose rotavirus antigens were positive and 65 negative ones were divided into two groups.The differences between these two groups in incidences of extra intestinal organ damage were analyzed.Meanwhile,variables from the clinical data that may lead to extra intestinal organ damage were analyzed.Then,the relationship of multiple organ damage and serum concentration of PCT was also analyzed.Results There were significant differences between positive group and negative group in the rates of respiratory system injury,myocardial damage and hepatic involvement (P ＜ 0.05).High fever was the only high risk factor in myocardial damage through multi factor Logistic regression analysis.There were also significant differences among the group with multiple organ damage and only one extra intestinal organ damage and no extra intestinal organ damage in serum concentration of PCT(P ＜ 0.05).Conclusion It is common to be attacked by extra intestinal organ damage in infants with acute rotavirus infection.High fever is the risk factor for RV enteritis complicated with myocardial damage.The elevation of PCT concentration suggest that multiple organ injury out of the intestinal tract may take place in infants with acute RV infection.

Aim To investigate and compare the phar-macokinetics of doxapram injection in healthy subjects of different Chinese nationalities including Han, Mon-golian, Korean, Hui and Uigur, and the influence of gender,in order to provide instruction and help for the usage of doxapram for both clinic and remedy of battle wound. Methods An HPLC-UV method was used to determine the plasma concentration of doxapram. Fifty healthy subjects ( five males and five females of each nationality) were recruited for the study. A single dose of 50 mg doxapram was administered intravenously to the healthy subjects, and blood samples were collected at various predetermined time points. The pharmacoki-netic parameters were calculated by DAS software and were compared by SPSS 13. 0 software, in order to as-sess the influence of nationality or gender on pharmaco-kinetics of doxapram. Results The results indicated that the pharmacokinetic profile of doxapram in vivo could be described as two-compartment model. The main pharmacokinetic parameters for Han, Mongolian, Korean, Hui and Uygur were as follows: Cl ( 0. 25 ± 0. 11 ) , ( 0. 33 ± 0. 11 ) , ( 0. 27 ± 0. 07 ) , ( 0. 26 ± 0. 06) and (0. 39 ± 0. 25) L·h-1 ·kg-1 , while Cmax (1. 55 ± 0. 52 ) , ( 1. 02 ± 0. 30 ) , ( 1. 31 ± 0. 47 ) , (1. 48 ± 0. 46 ) and ( 0. 99 ± 0. 35 ) mg · L-1 . The AUC0-12. 5 , AUC0-∞ and Cmax of Chinese Han were sig-nificantly higher than those of Uigur and Mongolian ( P<0. 05 ) , while there was no significant difference in other parameters ( P>0. 05 ) . There were statistically significant differences in Vc , Vd and CL between young males and females ( P < 0. 05 ) . Conclusion The large inter-individual variation in the main pharmacoki-netics suggests the dosage of doxapram should be ad-justed for different nationalities for both clinic and rem-edy of battle wound.

Objective To evaluate the efficacy and safety of recombinant adenovirus-p53(rAdp53) injection combined with radiotherapy and hyperthermia in the treatment of unresectable advanced soft tissue sarcoma.Methods In this retrospective study, we evaluated 76 patients with unresectable advanced primary or recurrent soft tissue sarcoma treated in our hospital from November 2005 to November 2012.These patients received radiotherapy and hyperthermia with rAdp53(p53 group, n=41) or without rAdp53(control group, n=35).rAdp53((1-2)×1012viral particles each time, once a week, 8 times on average) was injected into the tumor or infused into the pelvic cavity.Radiotherapy (2 Gy each time, 5 times a week) was performed for the planning target volume at 56.3±5.3 Gy in the p53 group and 58.1±4.2 Gy in the control group, with no significant difference between the two groups (P>0.05).Superficial or deep thermotherapy was employed 8 times on average (twice a week).Clinical features, response rate, time to progression (TTP), overall survival (OS), and adverse events were compared between the two groups (P>0.05).The Kaplan-Meier method was used to calculate OS;the log-rank test was used for survival difference analysis and univariate prognostic analysis;the chi-square test was used for comparison of categorical data.Results At 2 months after treatment, the p53 group had significantly increased response rate (partial response+ complete response+ stable disease)(85% vs.54%, P=0.003) and local control rate (49% vs.23%, P=0.020) as well as prolonged TTP (12 months vs.5 months, P=0.010) and OS (48 months vs.31 months, P=0.049), as compared with the control group.No adverse events caused by radiotherapy and hyperthermia except transient fever were seen in the two groups.Conclusions Concurrent radiotherapy and hyperthermia combined with rAdp53 injection is effective and safe for patients with advanced soft tissue sarcoma.

Objective: To understand the current situation and associated factors of unintentional injury among preschool children in Shunyi District, and to provide reference for the development of unintentional injury intervention measures. Methods: Three kindergartens in Shunyi district were selected through stratified cluster sampling method, and all the parents were surveyed online by self-designed questionnaire. Results: The proportion of low, medium and high risk assessment of unintentional injury in preschool children were 59.5%, 37.5% and 3.0%, respectively. Risk of unintentional injury increased significantly with age and grade(χ 2=12.35, 12.70, P<0.05). The risk of unintentional injury in inter-generational care (3.7%) was higher than that in parental care(2.4%). The higher the education level of the primary caretaker and family income, the higher level of unintentional injury risk(χ 2=11.23, 14.10, P<0.05).There were significant differences in the risk for burning, poisoning, other accidental injury, prevention of accidental injury and total score of unintentional injury among children of different ages and classes(F=8.26，5.61，4.95，6.15，7.86；9.88，8.39，4.25，6.27，7.55，P<0.05). There was statistical significance in burning risk between boys and girls(t=-4.27, P<0.05). There was statistical significance in unintentional injury prevention between children of different residence(t=9.11, P<0.05). There were significant differences in behavior supervision among risk among children of different ages and grades(P<0.05). Multiple linear regression analysis showed that education level of primary caregivers (college:B=-2.66, 95%CI=-4.69--0.63; bachelor degree or higher:B=-3.80, 95%CI=-5.90--1.70), annual family income (B=-2.82, 95%CI=-4.80--0.84) were associated with unintentional injury risk of preschool children(P<0.05). Conclusion Health education of unintentional injury prevention among preschool children should focus on the primary caretaker with low education and low family income, which is crucial for prevention of children s injury.