Animals ; Animals, Newborn ; Disease Models, Animal ; Enterocolitis, Necrotizing ; chemically induced ; Female ; Male ; Platelet Activating Factor ; toxicity ; Rats ; Rats, Sprague-Dawley ; Sulfalene ; toxicity ; Tumor Necrosis Factor-alpha ; toxicity

Animals ; Carcinoma, Hepatocellular ; genetics ; virology ; Hepatitis B, Chronic ; complications ; Hepatitis C, Chronic ; complications ; Humans ; Liver Neoplasms ; genetics ; virology ; Neoplasm Recurrence, Local ; genetics ; Point Mutation ; Telomerase ; metabolism

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.

Objectives To assess the prevalence of and risk factors for some sebaceous glands diseases (seborrhoeic dermatitis, acne, seborrhoeic alopecia and rosacea) and their relation to gastrointestinal dysfunction in adolescents. Methods A questionnaire survey was carried out for the epidemiologic information of sebaceous glands diseases from September to October 2001. A total of 4 600 youngsters aged from 12 to 20 years were recruited from 7 middle schools in Macau by random cluster sampling. The statistical software of SPSS 10.0 was used to analyze the data. Results The prevalence of seborrhoeic dermatitis, acne, seborrhoeic alopecia and rosacea was 2.7%, 45.5%, 1.6% and 0.7% respectively. Based on multivariate logistic regression analysis, the risk factors of sebaceous glands diseases included age, gastrointestinal dysfunction, season (summer), spicy and fried food, a family history of acne and the presence of long hairs on areolae of breasts. Gastrointestinal dysfunction was correlated with the incidence rates of seborrhoeic dermatoses (P = 0.007, OR = 1.208, 95% C.I 1.053 ~ 1.385). Conclusion Gastrointestinal dysfunction is an important risk factor for diseases of the sebaceous glands and correlates with the occurrence and development of these diseases.

Objective:To analyze the polymorphism of urease B gene(ureB) from different Hp strains. Methods:The sequences of ureB gene from 14 different Hp strains were amplified by PCR to analyze PCR-RFLP with HaeⅢ digestion.At the same time,the sequence of ureB gene was analyzed with biochemical software to compare the homology of nucleotide and amino acid sequence and to profile the phylogenetic tree. Results: The results of 1.7 kb ureB gene digested with HaeⅢ showed there were 2-5 band types in 14 different Hp strains and formed five distinct RFLP types.The two reference strains had the same RFLP type as 3 clinical isolates while the other clinical isolates and 3 isolates from animal model belong to 4 different RFLP types respectively.The nucleotide sequences and putative amino acide sequences of Hp ureB were compared.The various ureB sequences had high homologies(more than 96.6% and 98% in nucleotide sequences and amino acide sequences respectively) among 14 Hp strains.Particularly,there was the highest homologies(100%) between CCS9801、CCS9806、M3 and M10.Phylogenetic tree analysis showed that two reference strains and other isolates from clinical patients and Hp-infected mice model were located in two different lineage respectively in phylogenetic tree of nucleotide sequences while there were some variance in phylogenetic tree of amino acide sequences. Conclusion: Hp ureB was high conserved and homologous in the gene level as well as in the protein level.

OBJECTIVETo investigate the impact of in utero exposure to di-n-butyl phthalate (DBP) on the apoptosis of testicular cells in the pubertal male rat offspring.

METHODSTen pregnant SD rats were randomly divided into a control and an experimental group to be treated intragastrically with olive oil (1 ml per day) and DBP (500 mg per kg of body weight per day) respectively between gestation days 12 and 19. At the pubertal age (postnatal day 45, PND 45), the testes of the male rat offspring were removed for observation of the cell structure under the transmission electron microscope and the development of different spermatogenetic cells by HE staining. The apoptosis of testicular cells was detected by the TUNEL method, the expressions of the apoptosis-regulating proteins Bcl-2, Bcl-XL, Bax and p53 were determined by immunohistochemistry and Western blot, and the data obtained were compared between the two groups by t-test.

RESULTSTransmission electron microscopy revealed increased apoptosis and vacuolization of testicular cells in the PND-45 rat offspring, HE staining showed markedly decreased numbers of different spermatogenetic cells, TUNEL manifested significantly increased apoptosis of testicular cells in the experimental group as compared with the control (12.00 ± 5. 22 vs 3.17 ± 1.47, P < 0.01), and immunohistochemistry and Western blot exhibited remarkably higher expressions of Bax and p53 in the former than in the latter group (P < 0.05).

CONCLUSIONIn utero exposure to DBP can increase the apoptosis of germ cells and Sertoli cells, induce the vacuolization of testicular cells, and significantly elevate the expressions of the apoptosis-promoting proteins Bax and p53 in the pubertal male rat offspring.

Animals ; Apoptosis ; Body Weight ; Dibutyl Phthalate ; adverse effects ; Female ; Immunohistochemistry ; In Situ Nick-End Labeling ; Male ; Pregnancy ; Prenatal Exposure Delayed Effects ; Rats ; Rats, Sprague-Dawley ; Sertoli Cells ; cytology ; pathology ; Spermatogenesis ; Testis ; cytology ; pathology ; Tumor Suppressor Protein p53 ; metabolism ; bcl-2-Associated X Protein ; metabolism

Objective To observe the influence of different doses of intravenous immunoglobulin (IVIG) on function of T cells in cord blood of premature infants and to explore the immunomodulating mechanism of IVIG.Methods Cord blood mononuclear cells (CBMC) were isolated by density gradient centrifugation method from 15 preterm infants born between June 1,2011 to December 31,2011 at 32 34 gestational weeks.Three groups were formed according to the concentrations of IVIG used for CMBC culturing in vitro (Group A,6 mg/ml; Group B,9 mg/ml; Group C,12 mg/ml).After 72 hours in sterile conditions,the levels of IFN 7,IL 4,IL-10 and TGF β1 in the supernatant were determined by enzyme-linked immunosorbent assay.The expression of CD4+ CD25+ Foxp3+ Treg cells was examined by fluorescent activated cell sorter.Differences among groups were compared by one way analysis of variance.For comparison between two groups,LSD test was used.Results (1) The level of IFN γ secreted by CBMC in Group C was 0.03 ± 0.02,which was much lower than that in Group A(0.18±0.08) and Group B(0.13±0.05) (F=5.72,both P＜0.01).The level of IFN-γ in Group B and Group A did not show statistical difference (P＞0.05).Compared with Group A(0.03±0.01),the level of IL4 was much lower in Group B (0.02±0.01) and Group C(0.01±0.01) (F=20.38,both P＜0.01),while no significant difference was shown between Group B and Group C (P＞0.05).(2) No significant difference was found in the expression of CD4+CD25+Foxp3+Treg cells among different groups (F 0.67,P＞0.05).(3) The level of IL 10 in Group C was 3.02 ± 3.79,which was significantly lower than that in Group A (10.78±5.44) and Group B (6.90±4.64)(F=4.68,P＜0.01 and 0.05).The level of IL-10 inGroup B was still lower than that in Group A (P＜0.05).The levels of TGF-β1 in Group C(8.44± 13.71) and Group B(16.15 ±13.94) were significantly lower than that in Group A(30.23 ± 16.32) (F=5.22,P＜0.01,P＜0.05),but there was no significant difference between Group B and Group C (P＞0.05).Conclusions IVIG might inhibit the function of Th cells in CBMC of preterm infants in a dose dependent manner.IVIG could inhibit the function of natural Treg cells by regulating the secretion of cytokines IL-10 and TGF-β1 in a dose dependent effect.

Objective To induce mouse embryonic stem(ES)cells to differentiate into insulin-secreting cells by means of a 5-step model system.Methods E14.1 mouse ES cells were cultured in the presence of leukemia inhibitory factor(LIF)for 2 days(step 1),then the cells were cultured in hanging drops to form embryonic bodies(EBs)and the resulting EBs were cultured in suspension for 6 days in the presence of basic fibroblast growth factor bFGF(step 2).Subsequently the EBs were cultured in the medium containing glucagon- like peptide 1(GLP-1),hepatocyte growth factor(HGF),nerve growth factor(NGF)and nicotinamide for 10 days(step 3).After that,the EBs were dissociated into single cells,and the cells were cultured in monolayer in the presence of GLP-1,betacellulin,activin A,bFGF and nicotinamide for 10 days(step 4).Finally,the cells were cultured in low-glucose medium containing nicotinamide for 4 days(step 5).Insulin and some other islet- related genes expressions were investigated using RT-PCR and insulin expression was also investigated by DTZ- staining and immunohistochemistry.The percentage of insulin-secreting cells was evaluated by flowcytometry and insulin concentrations were measured by RIA.Results mRNA expression of insulin became visible at step 3 and more evident at step 5.Additionally,at step 5,mRNAs of glucagon,somatostatin,pancreatic polypeptide(PP), pancreatic duodenal homeobox 1(PDX-1),beta-cell E box transactivator 2(Beta2)and neurogenin 3(Ngn3) were detected.DTZ-staining positive cells and insulin immunohistochemical staining positive cells were observed. The percentage of insulin-positive cells was(24.0?2.5)%(n=6).In the presence of 5.6 mmol/L and 25 mmol/L glucose,insulin concentrations were(0.05?0.01)?g/L and(0.13?0.02)?g/L respectively(n= 6).Conclusion E14.1 mouse ES cells can be induced to differentiate into insulin-secreting cells by the 5-step model system.Insulin-secreting cells can release insulin into culture medium when treated with glucose,and insulin concentrations increase with rising concentration of glucose.

Objective To investigate the effects of metabolic syndrome(MS)on the structure and function of artery in patients with essential hypertension(EH).Methods One hundred forty-eight patients with EH were classified as EH with MS(EH+MS)or EH alone and healthy subjects(n=30)as control.Carotid intima-media thickness(IMT),brachial artery flow-mediated vasodilation(FMD)and non-flow-mediated vasodilation(NMD)were measured by Dopple ultrasonography.Results Carotid IMT(EH+MS:1.0?0.3 vs EH:0.8?0.2 vs control: 0.5?0.1 mm)and hsCRP(EH+MS:0.9?0.4 vs EH:0.7?0.3 vs control:0.4?0.2 mg/L)were significantly higher in EH+MS patients than that in EH+nonMS patients and control subjects(P all

Objective　To recombine and express a component of urease B subunit transmembrane protein of helicobacter pylori. Methods　A 732 bp gene fragment of urease B subunit of helicobacter pylori was cloned into pET11C and transformed into BL21（DE3）E.coli. The positive clone was induced with IPTG. The expression of target protein was analysed by SDS-PAGE and Western blot. Results　It is successful to construct the recombinant plasmid pET-UreB0.7 containing urease B subunit 0.7 kb gene fragment. A protein （MW≈28 000 u） with immunoreactivity， was expressed by 19.8％ in BL21（DE3）E.coli induced with IPTG. Conclusions　The recombinant component of urease B subunit transmembrane protein may play a role in the research of its biological function and might be used as the vaccine against helicobacter pylori.