0.05);the epididymal indexes were significantly increased 6,24 and 72 hours after formalin injection(P

This paper analyzed the moral construction in the prenatal diagnosis of surgical nursing and nursing features, including:strictness and complexity , professional and comprehensive , collaborative, and independence , carry out from the patients with preoperative evaluation , operation and postoperative effect evaluation every link and detail, with the theory of nursing ethics to guide and standardize nursing , dealing with the relation between various ethic, improve patient satisfaction , based on the premise of mutual modern relationship between nurses and pa-tients.

Objective To investigate the effect of pyrroloquinoline quinone(PQQ) on the aging of rat hippocampal neurons induced by D-galactose(D-gal).Methods Hippocampal neurons were cultured in vitro.The aging of the hippocampal neurons was induced by high dose D-gal,PQQ protection were used 30 min before D-gal.The metamorphosis of hippocampal neurons was observed under the microscope.The contents of free radical was measured.The incidence of apoptosis of hippocampus cells was tested with the flow cytometry.The expression of Bax was detected with immunohistochemical staining.Results After the cells cultured in vitro exposed to D-gal,the content of free radical and the expression of Bax of the hippocampal neurons increased.After pretreatment of the cultured neurons with PQQ,the contents of free radical and the expression of Bax decreased,the survival of hippocampal neurons increased.Conclusion PQQ may slow the aging progress of hippocamal neurons induced by D-gal.

Objective To investigate whether ROS/JNK/Egr-1 signaling pathway was activated in cardiomyocytes after hypoxia/reoxygenation ( H/R).Methods H9c2 cardiomyocytes were grouped randomly as follows: control group, H/R group, control +the ROS donor xanthine oxidase /hypoxanthine (XO/HX) group, H/R +the ROS scavenger edaravones (EDA) group, H/R +the ROS scavenger N-acetyl-L-cysteine (NAC) group, H/R +JNK inhibitor SP60012 group.To establish H9c2 H/R models and the myocardial cells were treated with different concentrations of EDA (2 × 10 -6,2 ×10 -5,2 ×10 -4 M), NAC (5 ×10 -4,2 ×10 -3,8 ×10 -3 M), XO/HX (1mU/mL/1.2 ×10 -4 M , 3mU/mL/3.6 ×10 -4 M, 5mU/mL/6.0 × 10 -4 M) and SP600125 (2 ×10 -5 M).ROS level was measured by flow cytometry, and Egr-1, p-JNK and total JNK protein levels were detected by Western blot.ResuIts ROS levels and Egr-1 protein levels in H/R group were significantly higher than control group (P<0.05).The moderate and high concentrations EDA and NAC of ROS scavenger significantly decreased the high levels of ROS and Egr-1 protein ( P<0.05 ) , but there were no significant differences of low concentration.There was a significant positive correlation between ROS levels and Egr-1 protein (r=0.91,P<0.01).JNK activation levels in each concentrations of XO/HX were significantly higher than control group, and JNK activation increased with the increasing of XO/HX concentrations (P<0.05).JNK activation level in H/R group was higher than control group, after treated by EDA and NAC of ROS scavenger and JNK inhibitor, JNA activation reduced (P<0.05).Egr-1 protein levels in H/R group was higher than that in control group, and JNK inhibitor reduced the expression of Egr-1 protein induced by H/R.ConcIusion H/R activates ROS/Egr-1 signaling pathway in H9c2 cardiomyocytes, and JNK activation plays an important role in this pathway.

Clinical data of 23 elderly patients with rectal cancer undergoing trans-sphincteric local resection(TSLR, Mason's operation)were retrospectively analyzed. All the 23 patients were followed-up for three to seven years after operation, 18 with normal or good fecal continence(78%), five just in fair condition(22%)and none in fecal incontinence. Three-year survival was 83 percent(19/23)and 5-year survival was 78 percent(18/23)for them. It is suggested that TSLR is a safe, feasible and effective treatment for middle and low rectal cancer in the elderly with a long term survival and satisfactory quality of life.

Objective:To construct eukaryotic expression plasmids of pneumocystis carinii p55-v3 and p55-v0 antigenic genes and to identify their expression in COS-7 cells at mRNA level.Methods:Pneumocystis carinii total RNA was used as the template to amplify p55-v3 and p55-v0 antigenic gene by RT-PCR.The products were connected to pTA2 vector and then cloned in pVAX1 eukaryotic expression vector to construct recombinant plasmids as pVAX-p55-v3 and pVAX-p55-v0.After propagated in E.coli DH5α,the recombinant plasmids were transfected into COS-7 cells.After 24 h incubation,the RT-PCR was performed to identify the mRNA expression of p55-v3 and p55-v0 antigenic gene.Results:The recombinant plasmids were qualified by restrictive endonuclease digestion and sequencing.And when compared with that in GenBank,the homology of p55-v3 antigenic gene was 99.9% in nucleotides and 100% in amino acid.The homology between p55-v0 antigenic gene and the one reported previously in nucleotide and amino acid seguence were 99.8% and 100%.The results of RT-PCR confirmed that p55-v3 and p55-v0 antigenic genes were transfected into COS-7 cells successfully and the genes were expressed in the cells.Conclusion:In this study,the recombinant plasmids of pVAX-p55-v3 and pVAX-p55-v0 are conducted successfully and expressed in the COS-7 cells,which provide a basis for clarification of immunologic function of p55-v3 and study of DNA vaccine.

Objective To investigate the anti-hepatocarcinoma(anti-HCC) function of HLA-A2-restricted point-mutated Survivin peptide induced CTLs.Methods The HLA-A2-restricted Survivin nonapeptides were evaluated using bioinformatics software.The binding affinity of Survivin peptide to HLA-A2 molecular was determined with flow cytometry analysis.After peptide-induced CTLs were generated in vitro,flow cytometry and ELISA were performed to detect the levels of IFN-γ,which were secreted by reactive CTLs.Peptide-induced CTLs were co-cultured with hepatoma cell lines HepG2 and BEL-7402.The rates of tumor cells lysis were assayed using CytoTox 96(R) and the morphological changes of tumor cells were observed with inverted phase contrast microscope.Results Point-mutated Survivin nonapeptide Sur79M2 (KMSSGCAFL) was filtered out,which was shown higher scores compared with the wild-type peptide Sur79.Consistent with the results of software analysis,Sur79M2 showed higher binding ability in T2 binding assays.At the same time,Sur79M2-induced CTLs could release a large number of IFN-γ after incubated with target cells rather than Sur79.When co-cultured with HCC cell lines HepG2 and BEL-7402,Sur79M2-induced CTLs effectively lysis HepG2 on HLA-A2-restricted manner without killing effect on BEL-7402 that do not express HLA-A2 molecules.Conclusion Sur79M2 could elicited specific cytotoxic T lymphocytes in vitro,which were able to specifically kill HCC cell lines on HLA-A2-restricted manner.

BACKGROUND:Embryonic stem cells (ESCs) serve as a major cell source for smooth muscle cells,but the heterogeneity of cells derived from ESCs result in difficulty to obtain high purity smooth muscle cells.OBJECTIVE:To construct a double expression vector of puromycin resistance (pac) gene and enhanced green fluorescence protein (EGFP) gene driven by smooth muscle specific SM22α promoter (pSM22α-PAC-IRES2-EGFP),in addition,to detect its availability and specificity in ESCs.DESIGN,TIME AND SETTING:The observational experiment of gene level was performed at the Cardiovascular Institute,General Hospital of Shenyang Military Region from April 2007 to September 2008.MATERIALS:ESCs line R1 with number SCRC-1011TM was purchased from American ATCC Company.The pSM22α-EGFP vector was constructed by our laboratory.And the pIRES2-EGFP,pSM2C and pSuper.basic vectors were purchased from Invitrogen Company.METHODS:SM22α promoter was cloned from pSM22α-EGFP by polymerase chain reaction.CMV promoter of pIRES2-EGFP vector was replaced by SM22 promoter to establish pSM22α-IRES2-EGFP.Pac gene,excised from pSM2C by HindⅢ/Clal digestion,was sub-cloned into pSuper.basic to establish pSuper-PAC.After BgⅢ/Accl enzyme digestion of pSuper-PAC,pac gene fragment was obtained,which was further sub-cloned into pSM22α-IRES2-EGFP to produce pSM22α-PAC-IRES2-EGFP.ESCs were transfected with pSM22α-PAC-IRES2-EGFP using lipofectamine.Positive clones were selected by G418 and induced to differentiate and further identified by amplification of pac gene by RT-PCR.Differentiated cells were immunostained by SM α-actin,and expression of SM α-actin and EGFP was observed simultaneously under fluorescence microscope.MAIN OUTCOME MEASURES:Sequencing result of pSM22α-PAC-IRES2-EGFP;Amplification of pac gene;EGFP expression;as well as SM α-actin immunostaining.RESULTS:Three segments of 261 bp,664 bp,and 5000 bp were obtained by HindⅢ/Clal digestion,which was coincident with expectation,and the sequencing results showed that pSM22α-PAC-IRES2-EGFP vector was successfully constructed.Amplification of pac gene identified 4 ESCs clones successfully transfected.After induction of differentiation,partial portion of differentiated cells expressed EGFP,accompanied by positively stained by SM α-actin antibody.CONCLUSION:pSM22α-PAC-IRES2-EGFP vector was successfully constructed.ESCs clones transfected with this vector expressed pac gene and EGFP gene,and the expression of EGFP is smooth muscle specific.

Objective To study the expression of gonadotropin releasing hormone receptor (GnRH receptor) in the stomach of Sprague-Dawley rat exposed to light stress. Methods We established illumination stressed models that the rats were exposed to continuous light(totally 64 rats, 32 rats for experiment and control group, respectively). Then,stomachs were taken from the rats when the rats exposed to continuous light for 1 day, 2 days, 3 days, 4 days, 1 week, 2 weeks, 3 weeks, 4 weeks and their control groups, respectively.The localization and the expression of GnRHR and GnRHR mRNA in stomachs mucous membrane were detected using immunohistochemistry, Western blotting and real-time PCR. Results Immunoreactivity was displayed mostly in the parietal cells in gastric gland. There were not differences of this distribution in all groups of constant illumination and their control. The immunoreactive materials were distributed on membrane and in cytoplasm of all positive cells, but not in nuclei. The level of GnRHR in rats exposed to continuous illumination were higher from one week to four weeks compared with that in control(P<0.05), and the level of GnRHR reached the peak when rats were exposed to constant illumination for two weeks (P<0.01). The quantity of GnRH mRNA in rats exposed to continuous illumination were higher from three weeks to four weeks compared with that in control(P<0.05). Conclusion The expression of GnRHR in digestive tract was effected by illumination stress.GnRH might regulate digestive function by interaction with GnRHR, suggesting that GnRH may be acted as a hormone, not only responded to normal physiological function of digestive tract but also responded to stress activity.

Objective To investigate the epidemiological and clinical characteristics, risk factors, outcome and prevention strategy of very low birth weight infant (VLBWI) with nosocomial infection in neonatal intensive care unit (NICU). Methods The VLBWIs whose birth weight were less than 1500 g and hospital stays were more than 48 hours in NICU of Peking University Third Hospital from January 1, 1998 to December 31, 2008 were selected in this study. They were divided into nosocomial infection group and non-infection group. The clinical features and outcomes of nosocomial infection were summarized and the risk factors of which were analyzed with Logistic regression. Results There were 158 VLBWIs who fit for the criteria of our study during the eleven years, the mean birth weight was (1263.8± 155.5) g and the mean gestational age was (30.4±2.1) weeks. There were 70 times and 56 cases suffered from nosocomial infections. The incidence of nosocomial infection was 35.4% and hospital stay-related incidence was 14.4‰. Among 70 times of infections, there were 40(57.1%) pneumonia, 22(31.4%) septicemia, 4(5.8%) thrush, 1(1.4%)conjunctivitis, 1 ( 1.4%) upper respiratory tract infection and 2 (2.9%) unknown site infections.Forty-one strains of bacteria were isolated from 121 specimens, among which gram-negative bacillus accounted for 56.1% and gram-positive cocci for 46.3%. The duration of hospital stay of VLBWIs with nosocomial infection was significantly longer than that without [(43.7±15.5) d vs (26.3±14.4) d] (t = -7.058, P＜0.01). The fatality rate of VLBWIs with and without nosocomial infection was 3.6% (2/56) and 3.9% (4/102), and there was no significant difference (x2 = 0.012,P＞0.05). Logistic regression showed that mechanical ventilation (OR = 3.388, 95% CI: 1.656-6.932, P=0.001) and parenteral nutrition (OR= 7.054, 95%CI: 2.005-24.813, P=0.002) were risk factors of nosocomial infection. Conclusions The incidence of nosocomial infection in VLBWIs in NICU is high. Mechanical ventilation and parenteral nutrition should be avoided and the duration of invasive operation and treatment should be shortened as much as possible to minimize the chances of nosocomial infection in VLBWIs.