Objective To observe the effects of fluoride of different dose on the proliferation of the cultured human umbilical vein endothelial cells(HUVEC). Methods The cells were treated with NaF at different doses. Cell counting, MTT colorimetry, flow cytometry (FCM) and immunohistochemistry were used to detect the endothelial cells activities. Results Compared with the fluoride free control, the positive rate of PCNA and Ki-67 were higher as NaF concentration was at 120-240 ?mol/L and were highest at 240 ?mol/L, the same was seen in the values of A and PI, as NaF concentration was at 600-960 ?mol/L,these indexes decreased. Conclusion In vitro, the low concentration of fluoride can promote the proliferative activity of HUVEC, whereas the high concentration can inhibit it.

Objective To evaluate the diagnostic value of high frequency echocardiography on mice with doxorubicin-induced cardiotoxicity and find out the most valuable monitoring indexes for early doxorubicin-induced myocardial damage.Methods Forty mice were divided into doxorubicin-injection group (ADM group) and control group randomly.Twenty mice in each group were injected with doxorubicin and saline at the same dose for 5 weeks continuously.The parameters of cardic function of ADM group and contol group were observed by high-frequency echocardiography after infection.Brain natriuretic peptide (BNP) tested by euzymelinked immunosorbent assay(ELISA) and pathological findings were used as the golden standard.Results ①The left ventricular long axis view and the apical four chamber view could be displayed clearly by transthoracic high-frequency echocardiography.②After 5 weeks intervention,weight,heart rate,left ventricular ejection fraction (LVEF),left ventricular fractional shortening (LVFS),E/A,E wave deceleration time(EDT) and other parameters showed marked difference between ADM group and control group(P ＜0.05).③The decrease of diastolic function was earlier than systolic function in ADM group.④LVEF and E/A presented good negative linear correlation to BNP(r =-0.622,P ＜0.05; r =-0.593,P ＜0.01).EDT showed positive linear correlation to BNP(r =0.501,P ＜0.05).Conclusions Cardic function of mice with doxorubicin-induced myocardial damage could be early evaluated accurately by high-frequency echocardiography,which contained good relativity with BNP.

Objective To evaluate the value of high-frequency echocardiography in assessing Bax gene transfer influence on survival of heterotopic cardiac allograft in rats.Methods Thirty rat models of heterotopic heart transplantation were established.Group A received heart transplantation only; Group B received cyclosporin (CsA) after operatiom Group C received ultrasound targeted microbubble destruction (UTMD) combined with Bax-shRNA.All rats were tested by high-frequency echocardiography at day 1,3,6 after transplantation.The ultrasound parameters included left ventricular internal dimension diastole (LVIDd),left ventricular internal dimension systole(LVIDs),left ventricular ejection fraction(LVEF),left ventricular thickness (LVT),left ventricular thickening (LVTR) and so on.The pathologic examinations were carried out on five rats at day 6 after echocardiography exam.The other rats in each group were observed for the survival time of cardiac allograft.Results ①The left ventricular long axis view and the apical four chamber view could be displayed clearly by high-frequency echocardiography.②The survival time of allografts in group C was (16.21 ± 5.01)d,which was longer than that in group B [(11.14 ± 1.72)d,P ＜ 0.05] and group A [(7.26 ± 1.50)d,P ＜0.01].③LVT index got higher after transfection.At day 6,LVEF of group A was lower than group B and C markedly(P ＜0.05) while no significant difference was found between group B and C(P ＞0.05).At day 3 and 6,LVT of group A and B were higher than group C (P ＜0.05) while LVTR was lower(P ＜0.05).Conclusions The Bax gene transfer influence on survival of heterotopic cardiac allograft in rats could be evaluated accurately by high-frequency echocardiography which could assess cardiac structure and function.LVT and LVTR can be considered as early evaluation indexes for its high sensitivity over LVEF.

OBJECTIVETo observe the effect of nitric oxide (NO) on the survival of a random pattern skin flap.

METHODSCaudal based random skin flaps (9 cm x 3 cm) were raised on the back of Wistar rats. Six methods were used in the experiment to observe the effect of NO synthase inhibitor L-NAME and NO synthase substrate L-arginine on flaps: image analysis technology; light and electron microscopic studies; enzyme histochemistry of NOS in flaps; concentration of NO2-/NO3- in plasma and wet/dry ratio of the flap tissue.

RESULTSSurvival area of flap in the L-arginine-treated group significantly increased (67.06 +/- 5.65)% (p < 0.01) whereas the area in the L-NAME-treated group significantly decreased (35.17 +/- 1.87)% (p < 0.01) compared with the control group (53.25 +/- 3.24)% at seven days after the operation. General and microscopic observations showed that pathological changes in the L-arginine-treated group were fewer. Abundant capillaries and fewer inflammatory cells were noticed in the L-arginine-treated group. Transmission Electron Microscopy (TEM) studies find endothelial swelling, thrombosis-formation and endothelial loss of contact with the basement membrane in the L-NAME treated group. Before operation, the serum NO concentrations were not significantly different in three groups (p > 0.05). After operation, NO concentration of the control group began to increase and reached to the top at the third day. L-Arg kept serum NO concentration in a higher level than the control. Enzyme histochemistry of NOS in flaps: microvessel intima in dermis, hair follicles, sweat glands and inflammatory cells showed oxford blue, more positive in flaps of the L-Arg treated group than the control group at the third day after operation. The flaps of L-NAME-treated group demonstrated negative or weak positive. Wet/dry ratio: twenty-four hours after flap elevation wet/dry weight ratios increased significantly in all regions of the flap of the L-arginine-treated rats compared with saline-treated rats. The ratios of the flaps of L-NAME-treated rats were reduced compared with saline-treated rats.

CONCLUSIONNO could improve microcirculation of the flap and increase its survival rates. The mechanism might be that NO could accelerate flap vascularization and protect flaps from ischemia-reperfusion injury.

Animals ; Arginine ; pharmacology ; Dermatologic Surgical Procedures ; Enzyme Inhibitors ; pharmacology ; Female ; Graft Survival ; drug effects ; Immunohistochemistry ; Male ; Microscopy, Electron ; NG-Nitroarginine Methyl Ester ; pharmacology ; Nitrates ; blood ; Nitric Oxide ; metabolism ; Nitric Oxide Synthase ; antagonists & inhibitors ; metabolism ; Nitrites ; blood ; Rats ; Rats, Wistar ; Skin ; enzymology ; ultrastructure ; Skin Transplantation ; Surgical Flaps ; physiology

Objective To silence Bax gene in H9c2 cells by ultrasound-targeted microbubble destruction(UTMD) combined with polyethylenimine.Methods The shRNA expression plasmid targeting Bax gene was co-treated with microbubbles and polyethylenimine.Then the mixture added to H9c2 cells were irradiated by ultrasound.Grouping:① plasmid group;② Lipofectamine2000 + plasmid group; ③ UTMD + plasmid group;④ PEI + UTMD + plasmid group;⑤ PEI + SonoVue + plasmid group (without ultrasound irradiation).The transfection efficiency was assessed by fluorescence microscope and FCM.The expression of Bax mRNA and protein were detected by RT-PCR and Western Blot.Results Bax shRNA wasn't destroyed by ultrasound exposure.The transfection efficiency of PEI + UTMD + plasmid group was higher than any other groups obviously (P ＜0.05).The result of RT-PCR and Western Blot showed inhibition efficiency of Bax mRNA and protein expression in PEI + UTMD + plasmid group were (69.41 ± 4.91) ％ and (64.09 ± 2.38)％ separately,which were higher than any other groups significantly (P ＜0.05).Conclusions UTMD combined with polyethylenimine can inhibit the expression of Bax gene in H9c2 cells effectively.

Objective To establish the reference ranges of the spatial angles among cardiac chambers and great vessels in second and third trimester fetuses measured by spatiotemporal image correlation (STIC).Methods Volume images of 352 normal fetuses from 20 to 38 weeks of gestation were recruited in the study.An off-line analysis of acquired volume datasets was carried out with multiplanar mode.Parameters measured included angles between:(1) the 4-chamber view and the left ventricular long axis view; (2) the left ventricular long axis view and main pulmonary artery; and (3) the ductal arch and aortic arch.The relationships between above-mentioned angles and gestational age were assessed by correlation and regression analysis.Results The angle between the 4-chamber view and the left ventricular long axis view (range:55.7° - 35.7°,mean:45.7° ± 5.12°) was uncorrelated with gestational age (r = 0.03,P = 0.51).In contrast,the angle between the left ventricular long axis view and main pulmonary artery,and the angle between the ductal arch and aortic arch were correlated with gestational age (P ＜ 0.001),and the correlation coefficient was - 0.53 and 0.57 respectively.The best-fit exponential curve regression equations of the angle between the left ventricular long axis view and main pulmonary artery was:Y = 154- 4.24X +0.05X2 ,and the angle between the ductal arch and aortic arch was:Y = - 20.8 + 2.65X - 0.37X2.Conclusions The angles among cardiac chambers and great arteries of fetuses from 20 to 38 weeks of gestation can be quantitatively measured by STIC.The reference ranges provide a reliable quantitative standard to estimate the spatial relationships of the cardiac large arteries of fetuses,which may be clinically useful in prenatal screening congenital heart disease.

OBJECTIVETo explore the genetic cause for a child with mental retardation, developmental delay and multi-systemic developmental disorders by analyzing the copy number variations (CNVs) and correlating the genotype with the phenotype.

METHODSRoutine G-banding was performed to analyze the karyotype of the patient and her parents. In addition, single nucleotide polymorphisms array (SNP-array) was used to determine the CNVs, which was confirmed by fluorescence in situ hybridization (FISH).

RESULTSNo karyotypic abnormality was detected upon chromosome analysis. However, SNP-array has identified a de novo hemizygous deletion of 1673 kb on chromosome region 7q11.23, which has been associated with Williams-Beuren syndrome. The microdeletion was confirmed by FISH testing.

CONCLUSIONA child with Williams-Beuren syndrome has been diagnosed by SNP-array and FISH. The de novo 7q11.23 microdeletion probably underlies the clinical manifestation of the patient. Compared with routine karyotype analysis, SNP-array is more useful for diagnosing children with multiple congenital anomalies with unclear etiology.

Adult ; Asian Continental Ancestry Group ; genetics ; Child, Preschool ; China ; Chromosome Banding ; Chromosomes, Human, Pair 7 ; genetics ; DNA Copy Number Variations ; Female ; Humans ; Karyotyping ; Male ; Pedigree ; Polymorphism, Single Nucleotide ; Williams Syndrome ; diagnosis ; genetics

【Objective】 To compare the bioactive ingredients in activated platelet-rich plasma (PRP) from cord blood and adult blood, explore its reasonable indicators reflecting the inflammatory regulation ability, in order to guide the preparation. 【Methods】 PRP was prepared and activated from 63 healthy adults (31 males, 32 females) and 61 neonates (30 males, 31 females), and 20 cytokines were measured using Luminex technology for assessing the age- and sex-based bioactive differences of PRP. High-sensitivity C-reactive protein(hs-CRP), procalcitonin and MMPs/TIMPs from each sample were measured for their correlations with the 10 inflammation-related cytokines. 【Results】 The activated cord blood PRP released 10 growth factors and chemokines more than the adult blood PRP, whereas IGF-1, HGF and 8 pro-inflammatory cytokines lower than the latter. Most cytokines of adult PRP were more in females than in males (P<0.05), except for IGF-1 and HGF, which showed no difference by gender. Compared with hs-CRP and PCT, MMPs/TIMPs ratio was more closely related with the inflammation-related cytokines, which can reflect the inflammatory regulation of PRP. 【Conclusion】 Due to the lower immunocompetence and no age or gender disturbance, cord blood PRP has the rational MMPs/TIMPs ratio and more cytokines which promote the inflammation and wound healing.

Objective:To explore the microbiological and disease distribution characteristics of multidrug-resistant bacteria in patients hospitalized in a critical care rehabilitation ward, and to analyze the risk factors leading to multidrug-resistant bacterial infections.Methods:Microbiology screening data describing 679 patients admitted to a critical care rehabilitation ward were retrospectively analyzed to divide the subjects into a multidrug-resistant group (positive for multidrug-resistant bacterial infections, n=166) and a non-multidrug-resistant group (negative for multidrug-resistant bacterial infections, n=513). The risk factors were then analyzed using logistic regression. Results:Among 369 strains of multidrug-resistant bacteria observed, 329 were gram-negative bacteria (89.2%), mainly Pseudomonas aeruginosa, Klebsiella pneumoniae and Escherichia coli. They were distributed in sputum (56.9%) and mid-epidemic urine (28.2%) specimens. Patients whose primary disease was hemorrhagic or ischemic cerebrovascular disease accounted for 40.96% and 23.49% of the multidrug-resistant bacterial infections, respectively. Logistic regression analysis showed that albumin level, dependence on mechanical ventilation, central venous cannulation, or an indwelling urinary catheter or cystostomy tube were significant independent predictors of such infections.Conclusion:The multidrug-resistant bacterial infections of patients admitted to the critically ill rehabilitation unit are mainly caused by gram-negative bacteria. Their occurrence is closely related to low albumin levels and mechanical ventilation, as well as to bearing an indwelling central venous catheter, a urinary catheter or a cystostomy catheter.

Abstract Allergic diseases can occur in all systems of the body, covering the whole life cycle, from children to adults and to old age, can be lifelong onset and even fatal in severe cases. Children account for the largest proportion of the victims of allergic disease, Children s allergies start from scratch, ranging from mild to severe, from less to more, from single to multiple systems and systemic performance, so the prevention and treatment of allergic diseases in children is of great importance, which can not only prevent high risk allergic conditions from developing into allergic diseases, but also further block the process of allergy. At present, there is no consensus on the management system of allergic children in kindergartens and primary schools. The "Consensus on Allergy Management and Prevention in Kindergartens and Primary Schools", which includes the organizational structure, system construction and management of allergic children, provides evidence informed recommendations for the long term comprehensive management of allergic children in kindergartens and primary schools, and provides a basis for the establishment of the prevention system for allergic children.