Objective To analyze the gene expression profiles in response to ΔNp63α overexpression, and screen the potential target genes or signal pathways regulated by ΔNp63α. Methods To generate ΔNp63α overexpressed SiHa cells ( SiHa-ΔNp63α) and the control cells ( SiHa-NC) , recombinant lentivirus transfection was performed. Microarray was applied to detect the change of gene expression profiles, and the results were analyzed with bioinfor-matic software. Quantitative real-time PCR was used to validate the expression levels of selected genes. Results Among the 1405 differentially expressed genes which were statistically significant, >1. 5 fold increase or reduce of gene expression, 843 were up-regulated and 562 were down-regulated in SiHa cells with ΔNp63α overexpression. The genes were mostly involved in cell development,cycle regulation, signal transduction, communication, adhe-sion, metastasis and invasion, etc. The involved signal pathways consisted of antigen processing and presentation, cytokine-cytokine receptor interaction, cell adhesion, complement and coagulation cascades, and so on. Conclu-sion The research on the potential target genes or mediated signal pathways regulated by ΔNp63α could be helpful to explain the development of cervical cancer.

Objective To explore the application of three-dimensional (3D) ultrasonography in screening the defects of fetal brain and skull.Methods Thirty-one cases of the fetal were detected in 3D ultrasonography.The ultrasonic data were compared with magnetic resonance imaging(MRI) and pathology.Results Among 31 cases according to 3D ultrasonography,there were 2 of anencephaly, 1 of exencephaly,2 of encephalocele,5 of holoprosencephaly,2 of arachnoidcyst, 1 of vein of Galen aneurysm,2 of agenesis of the corpus callosum, 1 of schizencephaly, 3 of Dandy-Walker syndrome, 7 of ventriculomegaly, and 5 of enlargement of cisterna magna.Compared with the results of MRI,the diagnostic accordance rate was 100% by 3D,it was 93.5% by 2D.Conclusions The structure of fetal brain and skull were viewed more clearly in 3D ultrasonography than 2D ultrasonography and the defects of fetal brain and skull can screened more exactly.

Objective To explore the effects of knocking down ULF gene on the apoptosis of non-small-cell carcinoma H1299 cell after treatment with etoposide.Methods Three ULF small interfering RNA(siRNA)sequences and one negative control siRNA sequence were designed and synthesized, and then individually transfected into H1299 cell via lentivirus.The interference efficiency of ULF-siRNA were screened by real-time PCR and Western blotting.Then the most target siRNA was used for apoptosis assay after treatment with etoposide,MTT assay for H1299 cell proliferation,flow cytometry for cell cycle distribution. Results The expression of ULF gene and its protein ULF were down-regulated in H1299 cell when transfected with ULF-siRNA,and ULF-siRNA-1 was the most effective one,which had the highest inhibition rate(80%)of ULF expression.Compared with negative control group,ULF-siRNA group showed an obvious apoptosis after treatment with etoposide,and the inhibition rate of was higher than control group,which was positively correlated with etoposide dose,the difference was statistically significant(P<0.05 ).Flow cytometry showed that compared with the control group,G0/G1 cell cycle in ULF-siRNA group was increased,and S phase cells was decreased,the differences were all statistically significant(P<0.05).Conclusion Down-regulation ULF protein expression through treatment with etoposide can induce apoptosis of non-small-cell carcinoma H1299 cells,and inhibit cell proliferation,which lead to cell cycle arrest.ULF gene may become the new target of gene therapy for cancer.

The knock out vector pHK was constructed with E coli vector pET 28a and shuttle vector pHY300PL, by using denatured DNA and homologous recombination technique, the kanamycin resistance gene ( Kan r) from integrated alkaline protease gene engineering strain BP071 was knocked out successfully, and the 11 positive clones were obtained The yield of the best positive clone BP0715 was stable as same as BP071 The methods provided the good experience for the industrial microbiology research, and it was foundation for studying on the safety of genetically modified organisms

Objective To investigate the prevalence and risk factors of renal injury in chronic hepatitis C patients(CHC).Methods The clinical data of 213 CHC patients,who were hospitalized in the People's Hospital of Peking University from Jan.2002 to Oct.2007,were collected.The eGFR was caculated by MDRD equation.The prevalence and risk factors of renal injury in the CHC patients was analyzed by SPSS software.Results The patients has an average age of (53.5?14.7)years old,with male patients accounting for 59.6% and female accounting for 40.4%.We also found that 22.1% patients had hypertension,25.8% had diabetes mellitus,and 94.8% had serum positive HCV RNA.The prevalence of CKD was 26.3%,the prevalence of proteinuria was 14.6%,and the rate of hematuria was 2.8%.Serumpostive HCV RNA was the independent risk factor of proteinuria as demonstrated by multiple variation logistic regress analysis(P=0.028,OR:2.610,95%CI:1.107~6.151).Proteinuria(P=0.02,OR:3.759,95%CI:1.227~11.521),age(P=0.004,OR:1.058,95%CI:1.018~1.100)and blood uric acid(P

Objective To study the clinical characteristics and prognosis of interstitial lung disease (ILD) associated with polymyositis (PM) and dermatomyositis (DM). Methods The clinical data of 107 DM related ILD,and its frequency rate was 26.2%. Arthritis as the first symptom had a higher presenting rate in patients with ILD than in patients with non- ILD. Arthritis, dry cough and short of breath occurred more dyspnea presented in patients with DM-ILD, and there was serious myalgia and muscle weakness in patients More DM-ILD patients had high HBDH and AST than those in PM-ILD. High CK and CK-MB were more The 7 of 8 severe cases had DM-ILD in which 5 died of respiratory failure (death rate was 17.9% in PM/elevated ESR and CRP tend to complicate with ILD. The DM patients who have characteristic skin rashes and high AST level are prone to develop ILD. The PM patients who have high CK and CK-MB are susceptible to

Wuxistatin, a novel and potent statin, is converted from lovastatin by Amycolatopsis sp. CGMCC1149. In the bioconversion, lovastatin is firstly hydroxylated by a hydroxylase. To obtain the critical hydroxylase, a novel hydroxylase gene was isolated from Amycolatopsis sp. CGMCC1149 by Degenerate PCR and Self-Formed Adaptor PCR and expressed in Escherichia coli. BLAST sequence analysis revealed that the gene belonged to cytochrome P450 gene superfamily and could encode a 403-amino-acid protein with a molecular weight of 44.8 kDa. The secondary structure prediction result showed that this protein contained many typical functional regions of P450, such as oxygen binding site, ion-pair region and heme binding region. Meanwhile, a catalytic function verification system was constructed by NADH, ferredoxin and ferredoxin reductase which could catalyze lovastatin hydroxylation into the target product. These would be helpful for further studies in large-scale production of wuxistatin.
Actinomycetales ; enzymology ; genetics ; Amino Acid Sequence ; Butyrates ; metabolism ; Cloning, Molecular ; Cytochrome P-450 Enzyme System ; genetics ; metabolism ; Hydroxylation ; Industrial Microbiology ; Lovastatin ; metabolism ; Molecular Sequence Data

Objective To observe homing of mesenchymal stem cells (MSCs) in immune organs and inflammatory joints in collagen induced arthritis(CIA) rats. MethodsRats MSCs were isolated and expanded from bone marrow cells by density gradient centrifugation and adhering to the culture cell walls, and the phenotypes were assessed by flow cytometry. MSCs were labeled by PKH-26 and Brdu. Sixty-four rats were randomly divided into normal group and CIA group. Every 8 rats were sacrificed at 3, 11, 30, 42 days after transplantation of MSCs. At the end of the experiment, the specimens of thymus gland, spleen, ankle joints were exposed, fixed, decalcified, wrapped and cut into slices. Confocal laser scanning microscope and immunohistochemical method were used to observe migration and distribution of MSCs in different organs. Independent samples group t test with SPSS 12.0 software package was used for statistical analysis. ResultsIt was found that allogenic MSCs could stay in spleen, thymus gland and joints of CIA rats for a relatively long period (42days). Forty-two days after transplantation of MSCs, the average grey scale values of spleen and thymus gland in CIA group(37.5±8.8, 29.9±5.9 respectively) were significantly higher than the normal group(16.0±2.3,13.2±4.3 respectively), the average grey scale values of ankle joints in CIA group 78±8 was significantly lower than the normal group 93±14(P<0.05). ConclusionIt has been found that MSCs can stay in the injured tissue and organs preferentially.

Objective To investigate the transcriptional autoregulation of PhoP/PhoQ under different growth conditions in Yersinia pestis.Methods The entire promoter region of YPO1635 was amplified and cloned into the pRW50 vector containing a promoterless lacZ reporter gene.The recombinant LacZ reporter plasmid was transformed into the wild-type strain (WT) and the phoP mutant strain (ΔphoP),respectively,to measure the promoter activity (the β-galactosidase activity) of the target gene in WT and ΔphoP by using the β-galactosidase enzyme assay system.Total RNAs were extracted from WT and ΔphoP strains,and primer extension assay was employed to detect the promoter activity by examining the amount of primer extension products of YPO1635 in WT and ΔphoP.Results The LacZ fusion results showed that the transcription of YPO1635 was positively regulated by PhoP under L-TMH and brain-heart infusion(BHI) conditions,but it was not regulated in H-TMH medium.The primer extension assay detected two transcriptional start sites located at 90 and 118 bp upstream of the translation initiation site of phoP,named P1 and P2,respectively.Under low Mg2+ TMH conditions,the promoter activity of P1 rather than P2 was positively regulated by PhoP.Under high Mg2+ TMH conditions,the promoter activities of both P1 and P2 showed no obvious difference in the WT and ΔphoP strains.Under rich BHI conditions,both promoters were under negative control of PhoP.Conclusion Different autoregualtion patterns of PhoP/PhoQ under different growth conditions would help Y.pestis to quickly adapt to the changing living environment.

Objective To discuss the complications of percutaneous kyphoplasty (PKP) in treatment of osteoporotic compression fractures (OVCFs) of the thoracic and lumbar vertebrae,and to investigate their causes and prevention.Methods From Jan.2004 to Dec.2012,PKP or PKP combined with percutaneous vertebroplasty(PVP) was performed on 126 vertebrae in 112 thoracolumbar OVCFs patients.There were 21 males and 91 females.The average age was 69 years (ranging from 36 to 85).100 cases had single vertebra,10 cases had double vertebrae,and 2 cases had 3 vertebrae.The injured segments were identified by MRI before surgery,and the fractures of the vertebrae were observed clearly by X ray and CT scan.The causes of the complications were retrospective analyzed.Results Perioperative complication happened in 68 cases (54％).The leakage of the bone cement happened in 66 cases (52.4％):one of them had high fever after the leakage happened behind the post-wall of the vertebra body,and it disappeared after one week with the relevant treatment.There was no injury of the nerve.Most of the leakage happened in front of the vertebra body,few happened in the terminal plate,and few happened behind the vertebral plate.There was no pulmonary embolism.Toxic reaction of the bone cement wasn't found in this team.There was no artery or nerve injury.One temporary spinal cord injury happened,one case was punctured in a wrong vertebra,and balloon rupture happened in one case.One case didn't achieve pain release.No infection happened.Conclusion The bone cement leakage was the most common complication associated with PKP in treatment of OVCFs.The complication rate can be reduced if the operation indications are strict,the volume of the filling cementis is well controlled,and the operation is performed in experienced hands.