Objective To explore the perioperative experiences of congenital heart disease in infants.Methods From Jan.2000 to Aug.2006,109 patients with congenital heart disease were operated in our department,their clinical data were retrospectively collected and analyzed.The patients′ age ranged from 31 days to 3 years old (13.6 months).The body weight ranged from 2.1 to 16 kg (8.6 kg).Ninety-three patients were operated under hypothermic anaesthesia with cardiopulmonary bypass(CPB).Sixteen patients underwent deep thermal and low flow CPB.Ultrafiltration was used in 62 patients.Results There were 8 deaths and the operative mortality was 7.3%,4 cases caused by low output syndromeclos(LOS),3 cases caused by pulmonary hypertension and 1 case caused by lung intection.The morbidity was in 25 cases(22.9%),the main complications were LOS in 6 patients and respiratory complications in 18 patients,hydropericardium in 1 case,respectively.Conclusion To improve the operative and CPB technique,and to improve the skills of the postoperative managements of LOS and respiratory complications are the main points in the success of the cardiac operation in infants.

Objective To analyze the epidemiological features and influencing factors of human brucellosis in Qinghai province,and to provide scientific basis for prevention and control of brucellosis.Methods From 2006 to 2010,select the high incidence areas of brucellosis in Qinghai province and five counties(Henan,Dari,Tianjun,Ping'an and Haiyan counties) included in the “Central Subsidies to Local Public Health Special Fund Human Brucellosis Prevention and Control Projects” for the survey point,as well as high-risk employees from Qinghai Biological Pharmaceutical Factory were investigated.Combined with epidemiological questionnaire investigation [done according to the “National Human Brucellosis Surveillance Program(Trial)”],clinical symptoms and signs,confirmed human brucellosis patient were tested by intradermal allergy test,rose bengal plate agglutination test(RBPT) and standard tube agglutination test (SAT),in accordance with “Diagnostic Criteria and Principles of Management of Brucellosis” (GB 15988-1995) and“Diagnostic Criteria for Brucellosis” (WS 269-2007).Results Of 8368 serum samples detected,347 were RBPT positive,and the positive rate was 4.15％;5346 serum samples were tested by SAT,180 were positive,and the positive rate was 3.37％.In June 2009,112 employees in Qinghai Biological Pharmaceutical Factory were investigated on a follow-up survey,83 were RBPT positive,the positive rate was 74.11％; 58 were SAT positive,the positive rate was 51.79％.Eight of them were new cases and 4 were chronic brucellosis.Twenty five new cases were reported between 2006 and 2010.The peak incidence was from March to July.Most of the cases were herdsmen.ConclusionStrengthening animal quarantine,strengthening public education,and improving protection awareness,can effectively control the disease brucellosis.

Objective To explore the relationship between X - linked spondyloepiphyseal dysplasia tarda (SEDL) gene escaping X chromosome inactivation( XCI) and SEDL phenotype. Methods RT - PCR was performed on total RNA which was isolated from blood samples of patients, female carriers and controls. Patients and female carriers were selected from the pedigree with SEDL caused by the mutation (IVS2 - 2A→C) of the gene. cDNA was analyzed by polyacrylamide gelelectrophoresis(PAGE). Results PAGE data indicateed that female carriers expressed both normal and mutant SEDL mRNA,meaning the SEDL gene escaping XCI. Family investigation showed carrier females in the SEDL pedigree presented no symptoms. Conclusions The SEDL gene escaping X chromosome in-activation is firstly identified from human body. This may explain that carrier females present no symptoms.

The effect of amygdalin joint hydroxysafflor yellow A (HSYA) on the endplate chondrocytes derived from intervertebral discs of rats induced by IL-1beta and the possible mechanism were studied and explored. Chondrocytes were obtained from endplate of one-month SD rat intervertebral discs and cultured primary endplate chondrocytes. After identification, they were divided into normal group, induced group, amygdalin group, HSYA group and combined group. CCK-8 kit was adopted to detect the proliferation of the endplate chondrocytes. FCM was measured to detect the apoptosis. Real-time PCR method was adopted to observe the mRNA expression of Aggrecan, Col 2 alpha1, Col 10 alpha1, MMP-13 and the inflammatory cytokines IL-1beta. The protein expression of Col II, Col X was tested through immunofluorescence. Compared with the normal group, the proliferation of the endplate chondrocytes decreased while the apoptosis increased (P < 0.05). With down regulation of the mRNA expressions of Aggrecan, Col 2 alpha1 and up regulation of the mRNA expressions of Col 10 alpha1, MMP-13, IL-1beta (P < 0.05), the protein expression of Col II decreased while the protein expression of Col X increased. Compared with the induced group, amygdalin group, HSYA group, the combined group could inhibit the apoptosis and promote the proliferation (P < 0.05). They could increase the mRNA expressions of Aggrecan and Col 2 alpha1 while decrease the mRNA expressions of Col 10 alpha1, MMP-13 and IL-1beta (P < 0.05). They could also enhance the protein expression of Col II while reduce the protein expression of Col X. The effect of the combined group was significantly better than that of amygdalin and HSYA. Amygdalin joint HSYA could inhibit the degeneration of the endplate chondrocytes derived from intervertebral discs of rats induced by IL-1beta and better than the single use of amygdalin or HSYA.
Amygdalin ; pharmacology ; Animals ; Apoptosis ; Cells, Cultured ; Chalcone ; analogs & derivatives ; pharmacology ; Chondrocytes ; drug effects ; Collagen ; metabolism ; Drug Synergism ; Interleukin-1beta ; Intervertebral Disc ; cytology ; Quinones ; pharmacology ; Rats

Objective To investigate the effects of cryopreservation on the growth and osteogenesis capa- bility of human bone marrow stromal cells(BMSCs)on demineralized bene matrix(DBM).Methods Bone marrow aspirates were obtained from the lilac crests of three donors.The BMSCs were isolated from the bone marrow by density gradient centrifugation.Cells of passage 3 were cryopreserved in liquid nitrogen for 24 hours,and then re- covered.The non-cryopreserved BMSCs were used as the control,The cryopreserved and control BMSCs were cul- tured in osteogenic media,collected and labeled with Dil to be seeded onto the DBM when cells were confluent.The percentage of BMSCs adhered to the DMB was detected.The cell morphology and matrices secreted by BMSCs on the DBM were observed by the inverted phase-contrasted microscope,fluorescence microscope and scanning electron microscope(SEM).The growth and viability of BMSCs on the DBM were determined using the modified MTT ashy. The osteogenesis ability of BMSCs on the DBM was determined by assessment of the alkaline phosphatase(ALP) activity and osteocalcin(OCN)content.Results The percentages of the cryopreserved and control cells adhered to DBM were(97.25?1.17)% and(97.00?1.09)% respectively.The cells adhered well to the DBM and grew rapidly.Large amounts of matrices on the DBM were observed by the light microscope and SEM.The cells embedded in the matrices could be observed by fluorescence microscope.There were no significant differences in the assay values of MTT,ALP and OCN between the cryopreserved and control BMSCs on the DBM.Conclusion Since cryopreservation does not affect the growth and osteogenesis capability of BMSCs on DBM,the cryopreserved BMSCs can be used as a cell source in bone tissue engineering.

OBJECTIVEThis paper aims to develop a monoclonal antibodies (MAbs)- based ELISA for detecting Chlamydophila pneumoniae (C. pneumoniae) antigens in humans with the variable domains (VD) 2 and 3 of the major outer membrane protein (MOMPVD2-VD3) and to assess its sensitivity and specificity by comparing with a widely used MAb that is able to recognize the elementary bodies of C. pneumoniae.

METHODSMOMPVD2-VD3 were overexpressed in Escherichia coli and purified by affinity chromatography. Mice were immunized with the recombinant antigen, and hybridomas secreting MAbs were screened. Three stable hybridomas clones were selected and named 5D6, 7G3, and 8C9. The MAbs-based ELISA was scrutinized for species-specific recognition with a number of human throat swab samples from Group I (156 patients with typical respiratory illness clinically confirmed before) and Group II (57 healthy donors).

RESULTSIn Group I, 55 positive cases were detected by anti-EB MAb-based ELISA, 51 cases were positive by MAbs 5D6-based ELISA, and 33 and 38 cases were positive by MAb 8C9 and 7G3-based ELISA respectively. Of the 57 samples from Group II "healthy donors", 5 were positive and 52 were negative with both anti-EB and 5D6-based tests, while 2 and 3 positive cases were identified by the other two MAb-based ELISAs respectively.

CONCLUSIONThe novel MOMPVD2-VD3 MAb-based assay may have higher specificity than the anti-EB MAb, which may possibly be used as an alternative tool for the diagnosis of C. pneumoniae infection.

Animals ; Antibodies, Monoclonal ; Bacterial Outer Membrane Proteins ; immunology ; Chlamydophila Infections ; diagnosis ; microbiology ; Chlamydophila pneumoniae ; isolation & purification ; Enzyme-Linked Immunosorbent Assay ; methods ; Humans ; Mice ; Protein Structure, Tertiary

Objective To explore the relationship between dynamic changes in ultra-micro-structural of pulmonary arteries and endogenous hydrogen sulfide in rats with left-right shunt.Methods Rats in shunt group were subjected to an abdominal aorta-inferior vena cava shunt to create an animal model of pulmonary artery structural remodeling. After 1 day, 3 days, 1 week, 4 weeks and 8 weeks of experiment, the ultra-micro-morphologic changes of pulmonary arteries of rats were observed under electronic microscope and H_2S concentration in serum was evaluated by modified sulfide electrode method.Results The changes of ultra-micro-structure of pulmonary arteries were progressively exacerbated, endothelial cells became swollen and large in size on 3 days, smooth muscular cells increased in size as well as the change of endothelial cells in 1 week, and they changed from contractile phenotype to synthetic phenotype in 4 weeks.Conclusions Shunt exhibited changes of ultra-micro-structure of pulmonary arteries are accompanied by the changes of endogenous H_2S. It is suggested that endogenous H_2S might play a protective role in changes of ultra-micro-structure of pulmonary artery.

Objective To study the effect of dynamic stress stimulation on the expression of Sox9 mRNA and protein in metaphyseal chondrocytes in vitro, and to explore the specific mechanism of mechanical signal transduction. Methods The rat metaphyseal chondrocytes separated and cultured for the 3rd generation in vitro were randomly divided into four groups:control group (all interventions were not applied), simple dynamic pressure group (a dynamic pressure stimulus with a size of 90 mmHg and a frequency of 0.1 Hz was applied using an open pressure control culture system), simple calcium antagonist group (the concentration of 10μmol/L nifedipine was given) and dynamic pressure+calcium antagonist group (a dynamic pressure stimulus with a size of 90 mmHg, frequency of 0.1 Hz and concentration of 10 μmol/L nifedipine were given at the same time). The expression of Sox9 mRNA was detected after 24 h intervention by real-time quantitative polymerase chain reaction (RT-PCR) in four groups. The expression of Sox9 protein was detected by Western blot assay. The intracellular free Ca2+ in metaphyseal chondrocytes was labeled with Fluo-3/AM, and the average fluorescence intensity detected by laser scanning confocal scanning microscopy was compared between four groups. Results The expression of Sox9 mRNA was 3.81 times higher in dynamic stress group than that in the control group, and the protein expression level was 2.33 times higher than that of the control group (P<0.05). There were no significant differences in the expression of Sox9 mRNA and protein between the calcium antagonist group and the control group. The expressions of Sox9 mRNA and protein were lower in dynamic pressure+calcium antagonist group than those in the dynamic stress group, but which were higher than those of control group(P<0.05). The results of average fluorescence intensity showed that there was no significant difference in the intracellular free Ca2+ concentration between four groups (P > 0.05). Conclusion Dynamic stress stimulation can increase the expression of Sox9 mRNA and protein in rat metaphyseal chondrocytes. There is calcium channel involvement in the mechanical signal transduction.

Objective To study the immunogenicity of human embryonic stem cells(hESCs)and the derived neural stem cells(NSCs)in vitro.Methods The constitutive expression of human leucocyte antigen(HLA)Ⅰ and Ⅱ in hESCs and the NSCs derived from these hESCs were detected by flow cytometry (FCM), as well as the expression of HLA-Ⅰ,Ⅱin NSCs induced by 30 ng/ml recombination human interferon-?(IFN-?).Meanwhile, the NSCs before and after induction of IFN-? were co-cultured with peripheral blood lymphocyte obtained from healthy person.Lymphocyte proliferation standing for the immunoreactivity of NSCs was then investigated.Results The hESCs slightly expressed HLA-Ⅰ(6.18%) and hardly any HLA-Ⅱ before differentiation.However, the NSCs expressed more HLA-Ⅰ(23.56%)as well as HLA-Ⅱ(1.28%, 1.73%)than the hESCs did.Both HLA-Ⅰ(46.43%)and HLA-Ⅱ(8.73%, 10.57%)expressed by the NSCs after they were induced by IFN-? were up-regulated.Conclusions hESCs express certain level of HLA-Ⅰ molecules but do not constitutively express HLA-Ⅱ molecules.The derived NSCs express heavy HLA-Ⅰ and a little HLA-Ⅱ, when treated by IFN-? they can inducibly up- regulated both molecules.The NSCs derived from HESCs are of immunogenicity, which induce rejection aiming at HLA-Ⅰ molecules or even at HLA-Ⅱ molecules when the host is inflammative or under stress, which can result in a failure of cellular transplantation.

BACKGROUNDHypertension is a common disease of the cardiovascular system. So far, the pathogenesis of primary hypertension remains unclear. The elaboration of its pathogenesis is an important topic in the field which calls for urgent resolution. The aim of this study was to probe into the metabolic imbalance of homocysteine (Hcy) and hydrogen sulfide (H(2)S) in children with essential hypertension, and its significance in the pathogenesis of essential hypertension.

METHODSTwenty-five children with essential hypertension and 30 healthy children with normal blood pressure were enrolled in the study. The medical history was investigated and a physical examination was conducted on the subjects. Plasma Hcy content was examined by fluorescence polarization immunoassay (FPIA). The plasma H(2)S level was detected by a modified method with a sulfide electrode. Data were presented as mean +/- standard deviation. The t test was applied to the mean values of both groups. Pearson linear correlation analysis was applied to the plasma Hcy and H(2)S as well as to the systolic pressure against the plasma H(2)S/Hcy ratio.

RESULTSPlasma Hcy, an intermittent metabolite of the endogenous methionine pathway, was markedly increased but plasma H(2)S, a final product of this pathway was significantly decreased in hypertensive cases when compared with normal subjects ((Hcy: (12.68 +/- 9.69) micromol/L vs (6.62 +/- 4.79) micromol/L (t = 2.996, P < 0.01); H(2)S: (51.93 +/- 6.01) micromol/L vs (65.70 +/- 5.50) micromol/L) (t = -8.670, P < 0.01)). The ratio of plasma H(2)S/Hcy in children with hypertension was 5.83 +/- 2.91, while that of the control group was 11.60 +/- 3.30, and the difference is significant with a t = -6.610 and P < 0.01. A negative correlation existed between plasma Hcy and H(2)S concentrations, r = -0.379, P < 0.05. And a negative correlation was found between systolic blood pressure and the plasma H(2)S/Hcy ratio, r = -0.687, P < 0.05.

CONCLUSIONThere was a metabolic imbalance of homocysteine and hydrogen sulfide in essential hypertensive children.

Adolescent ; Child ; Female ; Homocysteine ; metabolism ; Humans ; Hydrogen Sulfide ; metabolism ; Hypertension ; etiology ; metabolism ; physiopathology ; Male ; Systole