Objective To explore optimal initial the best screening time for newborns with different delivery methods using AABR.Methods A total of 550 newborns who were born from August 1, 2016 to October 31, 2016 at our hospital participated in the study.AABR was used to accomplish the initial hearing screening.The newborns were divided into 2 groups according to the delivery methods.There were each 100 neonates born in vaginal during <24 h, 24~48 h and 48~72 h after birth, respectively.The numbers of neonates delivered by cesarean section during the 3 separate periods were 50, 100 and 100, respectively.The newborns who failed the preliminary hearing screening proceeded to the re-screening and diagnostic procedures.Results There were 300 newborns were born in vaginal, and the pass rate in 24~48 h after birth group was significantly higher than that in 24 h group (93.00% vs 83.00%,x2=4.735,P=0.03<0.05), but it was not significantly different from that of 48~72 h group (95.00% vs 93.00%,x2=0.355,P=0.56>0.05).There were 250 newborns in cesarean section, the pass rate of 24~48 h after birth group was significantly higher than that in 24 h group (83.00% vs 68.00%,x2=4.437, P=0.04<0.05), and significantly lower than that of 48~72 h group (94.00% vs 83.00%,x2=5.944, P=0.02<0.05).Conclusion Taking into account of hospitalization time, the screening time for the vaginal delivery newborn hearing screening can be advanced to 24~48 h after birth with the application of AABR, but not for the cesarean section group.

Objective To construct the expression vector and express the recombinant human L-selectin in prokaryotic system, and purify the aimed protein for the preparation of rabbit anti-human L-selectin polyclonal antibody. Methods The partial cDNA of human L-selectin was amplified from the open reading frame (ORF) of N-terminus sequence of L-selectin containing 153 amino acids by PCR, then cloned into the prokaryotic expression vector pQE40 at restriction sites BamH Ⅰ and Hind Ⅲ. The recombinant expression plasmid pQE40-L-selectin was transformed into E. coli M15 for expression. The fusion protein including 6-His-tag was purified by Ni-NTA chromatographic column and analysed by SDS-PAGE. The purified protein was used to immune rabbit for preparing polyclonal antibody. Western blotting analysis and dot immunoblot assay (DIBA) were used to test the titer of the antiserum. Results The expression plasmid pQE40-L-selectin was constructed and confirmed with restriction enzyme digestion. The quantity of the purified protein from E. coli M15 was 600 mg ? L-1. By immuning the rabbit, the polyclonal antibody was successfully prepared. The results of dot immunoblot assay (DIBA) showed that the antiserum had the high titer (1 : 1 000). Conclusion The recombinant human L-seleclin prolein can express with high efficiency in E. coli M15. The prepared polyclonal antibody has a high titer.

Objective To study the effects of resveratrol on cell morphology and related factors of gastric cancer cell line in vitro.Methods Drug sensitivity was detected by MTT assay.Changes of its biological characteristics were determined using light microscopy,electron microscopy cell counting by MTT assay,flow cytometry(FCM).Results Resveratrol(0.1g/L、0.2g/L)could arrest the suspended gastric cancer cell(MGC803)to S phase respectively.Resveratrol significantly inhibited growth and proliferation of MGC803 cells in dose-dependent manner.Conclusion Resveratrol(0.1g/L 、0.2g/L)may inhibit MGC803 cell growth.

Objective To investigate the mechanism of xCT on tumor metastasis in breast cancer cell MDA-MB-231. Methods Wound scratch assay and Transwell assay were performed to evaluate the effect of disruption and knockdown of xCT on cell migration and cell invasion in breast cancer cell MDA-MB-231 .Western Blot and RT-PCR were used to detect the expression levels of autophagy and EMT related markers in breast cancer cell MDA-MB-231 after treatment with sulfasalazine (SASP), an inhibitor of xCT activity and SLC7A11-RNAi.Results Both the scratch assay and the transwell migration assay showed that inhibition of xCT reduced the motility of MDA-MB-231 .The expression level of autophagy related protein LC3-Ⅱ/LC3-Ⅰwas elevated, the protein level of transcription factor Snail was down-regulated, while the mRNA level of Snail did not change in xCT inhibited MDA-MB-231 cells compared with MDA-MB-231 cells.Epithelial marker E-cadherin was up-regulated but mesenchymal marker Vimentin was down-regulated when xCT was deficient.Con-clusion Our current studies show that xCT is an endogenous regulator of tumor growth and metastasis in MDA -MB-231 and the expression level of xCT determines the phenotypes of MDA-MB-231 cells in invasion and migration in vitro.Inhibition of xCT can activate autophagy , induce the degradation of Snail ,and attenuate the EMT process in highly metastatic MDA-MB-231 cells.

BACKGROUND: Smooth muscle cells (SMCs) proliferation and transmigration and platelet activation cause thrombogenesis and lead to grafted vessel restenosis. Nitric oxide (NO) can inhibit the above-mentioned biological responses, but whether endothelial nitric oxide synthase (eNOS) gene transfection can inhibit the neointimal hyperplasia in graft seeded with SMCs remains uncertain.OBJECTIVE: This study was designed to further investigate the effect of eNOS gene transfection on neointimal hyperplasia in the grafts seeded with SMCs.DESIGN, TIME AND SETTING: This study, a repeated observation and measurement experiment, was performed at the Central Laboratory and Laboratory of Molecular Biology of Xi'an Jiaotong University Medical College from April 2006 to May 2007.MATERIALS: One 1-month-old New Zealand rabbit was used to acquire SMCs. Another 18 adult New Zealand rabbits were randomly divided into 3 groups (n=6).In normal control group,the vessel graft with no SMCs were transplanted; In SMC/lacZ group, the vessel grafts with SMCs transfected with lacZ were transplanted;In SMC/eNOS group,the vessel grafts with SMCs seeded with eNOS were transplanted.METHODS: Rabbit SMCs were transduced with pseudotyped retroviral vectors, Murine leukemia virus/vesicular stomatitis virus G glycoprotein, carrying genes coding for eNOS or lacZ gene. The SMCs then were seeded on the vessel grafts and implanted into the rabbit abdominal aorta using vessel bypass transplantation.MAIN OUTCOME MEASURES: Nitric oxide (NO) content in the supernatant of cells transfected with eNOS and lacZ gene was detected by citrulline method. The grafts were stained with X-gal to visualize the seeded cells: the seeded SMCs were stained blue,while eNOS were stained red. The thickness of the neointima on a graft was measured with a microscope.RESULTS: Eighteen rabbits were all included in the final analysis. NO content in the SMC/eNOS group was significantly higher than that in the normal control group (P<0.05). The SMCs transfected with lacZ gene showed blue after X-gal staining under the inverted microscope. Thirty days after implantation, there was no difference in neointimal thickness between normal grafts and grafts seeded with eNOS or lacZ transduced SMCs (P>0.05).100 days after implantation,the neointimal thickness on grafts seeded with eNOS transduced SMCs was similar to that of unseeded grafts (P>0.05 ), but was significantly thinner than that on grafts seeded with SMCs transduced with only lacZ gene (P<0.05).CONCLUSION: eNOS gene transfection inhibits nenintimal hyperplasia in the vessel graft seeded with SMCs.

Objective To construct pseudotyped retroviral vector MuLV/VSV-G and transfer it into rabbit smooth muscle cells (SMC) in order to provide a high-efficiency vector for SMC gene transfer. Methods We constructed pseudotyped retroviral vector MuLV/VSV-G containing the previously reported gene lacZ, determined the titer, and determined the efficiency of gene transfer into SMC mediated by pseudotyped retroviral vector MuLV/VSV-G. Finally the transfer efficiency was compared with that by MuLV. Results MuLV/VSV-G vector was constructed. The titer of the vector was 6-7.8×10~6CFU, the transfer efficiency was (92±12)% by using MuLV/VSV-G vector and (24±5)% by MuLV vector. Conclusion Pseudotyped retroviral vector MuLV/VSV-G which was constructed successfully is a kind of high-efficiency gene transfer vector in smooth muscle cells.

Objective To obtain the change of bcl-2/bax/fas/fasL in splenic lymphoctyes with different lasting time of hypoxicischemic brain damage (HIBD). Methods The newborn rat were divided into 6 groups by the time of being HIBD model randomly, includes 1/6/12/24/48/72 hour(s) (8 for every group),and control groups were established at the same time point. The following four apoptosis related genes bcl-2/bax/fas/fasL were tested by real time PCR. Results ( 1 ) bcl-2: the mRNA expressions of HIBD groups were lower than control groups at the same time ( P<0.01 ). Eliminated the control effects, the mRNA expressions of HIBD groups were differernt by the modeling time(P ＜0.01 ). (2)bax: the mRNA expressions of HIBD groups were higher than control groups at the same time( P ＜0.01 ), and in control group the expression of 6 h was much higher than any other groups (P<0.01 ). Eliminated the control effects, the mRNA expressions of H IBD groups were different by the modeling time( P<0.01 ). (3)bcl-2/bax: the ratios of HIBD groups were lower than control groups at the same time( P ＜0.05 ), the ratios in control groups were higher than 1 ( except for 1 h); while in HIBI) groups the ratios were lower than 1; Eliminated the control effects, the ratios were different in all the groups. (4)fas: the mRNA expressions of HIBD groups were higher than control groups at the same time ( P ＜0.01 ), and both were maximum at 6 h. (5)fasL: the mRNA expressions of HIBD groups were higher than control groups in 1 h and 6 h ( P<0.01 ), while lower than control group at other time points( P<0.01 ),the expression of 24 h was the maximum of control groups and 12 h was the maximum of HIBD groups. (6)fas/fasL: the ratios of HIBD groups were higher than control groups( P ＜0.01 ) (except for 6 h), and the ratios in control groups were lower than 1 ( P<0.01 ) ( except for 6 h), and not concentrated, while in HIBD groups were higher than 1 ( except for 24 h), between 0.69 to 5.65. Conclusion Pro-apoptosis genes ( include bax/fas/fasL) were promoted by HIBD, while anti-apoptosis gene(bcl-2) was inhibited. The maximum of pro-apoptosis genes became early in HIBD. Both the pro- and anti-apoptosis genes got their maximum at 6 h and 12 h of HIBD. The apoptosis suppression was the main effects in control groups from the ratio of bcl-2/bax, which was lower than 1. The apoptosis promotion was the main effects in HIBD groups from the ratio of bcl-2/bax, which was higher than 1, especially at 12 h. Thefas/fasL effect which is the major way of lymphocytes apoptosis was strengthened in HIBD.

Objective To culture preadipocytes in vitro and to study the cell compatibility of PLGA scaffolds,collagen scarfolds and hyaluronic acid-based scaffolds and tO choose the optimal seeding method.Methods The preadipocytes from human abdominal adipose tissue were isolated and cultured in enzyme-digesting method.The generation of human preadipocytes was planted on PLGA scaffolds,collagen scaffolds and hyaluronic acid-based scaffolds.and the cell compatibility was observed by MTT method.The seeding efficiency of human preadipocytes on scaffolds.human preadipocytes were seeded to hyaluronic acid-based scaffolds by static culture and stirred culture.Results Compared compatibility of preadipocyte with three different scaffolds,there was great difference between hyaluronic acid-based scaffolds and PLGA scaffolds.Difference also existed between hyaluronic acid-based scaffolds and collagen scaffolds that were different from PLGA scaffolds.Among them,hyaluronic acid-based scaffolds was the best.Conclusion Hyaluronic acid iS a better scaffolds material for adipose tissue engineering compared with PLGA and collagen.The seeding efficiency of stirred culture is higher than static culture,which is an optimal method for cell seeding tO 3-D scaffolds.

Objective To establish a rat model of liver metabolism profile in chronic heart failure (CHF), to explore the dynamics of liver metabolism in CHF from the point of view of metabolism, and to find the characteristic metabolites valuable for the molecular mechanism and management of CHF.Methods Twenty male Wistar rats were assigned to the CHF group to receive aortic coarctation or to the control group to receive sham surgery, and were bred for 24 weeks following surgery.The metabolic profiling of the rat liver tissues was analyzed on a metabonomics research platform. Orthogonal partial least squares-discriminant analysis ( OPLS-DA) model and principal component analysis ( PCA) model were established for liver tissues of the CHF rats, and the characteristic metabolites were finally derived by data processing with SPSS 19.0 software.Results The PAC and OPLS-DA models were established successfully.Ten characteristic metabolites with significant differences between the CHF and control groups, including lysophosphatidyl choline, lysophosphatidyl ethanolamine, oleic acid, glycocholic acid, and dehydroepiandrosterone sulfate, were screened and identified from the models.Conclusions The metabolic disorders in CHF rats are well fitted to the established metabolic profile models, and these identified characteristic metabolites may provide reference for the pathophysiological molecular mechanism and management, etc., of chronic heart failure.

Objective To investigate the genetic stability, immunogenicity and protective efficacy of AdC68-rab. gp, a novel rabies vaccine based on the replication-defective chimpanzee adenoviral vector AdC68-ept. Methods The recombinant adenovirus AdC68-rab. gp expressing the glycoprotein of rabies vi-rus ERA strain was constructed. Genomes of the AdC68-rab. gp of different generations were extracted and analyzed. HEK293 and Huh7 cells were infected with the AdC68-rab. gp of different generations. ICR mice were immunized with the AdC68-rab. gp and blood samples were collected 4 weeks or 6 months after immuni-zation. Rapid fluorescent focus inhibition test ( RFFIT) was performed to detect the neutralizing antibody against rabies virus in mice serum samples. ICR mice were challenged with lethal dose of rabies virus 4 weeks after the immunization with AdC68-rab. gp to evaluate the protective efficacy of AdC68-rab. gp. Re-sults The genome of AdC68-rab. gp was stable after 15 passages, which was identical to that of the 5th and 1st generations. High levels of neutralizing antibody against rabies virus in serum samples were detected in mice immunized with AdC68-rab. gp and maintained for a long period of time. Immunization mice with one dose of AdC68-rab. gp could protect all mice from the lethal dose challenge of rabies virus. Conclusion The novel AdC68-rab. gp was characterized by good genetic stability and ideal protective effi-cacy. The adenoviral vector based vaccine could be further developed as a potential candidate for the substi-tute of current rabies vaccine.