Objective To obtain an easy and high efficient method for gene transfer to cochlear hair cells ,by comparing three mediating green fluorescent protein (GFP) methods (electroporation ,adenovector and lentivirus vector) .Methods Cochlear sensory epithelium was dissected from anaesthetized P2 mice .Sensory epithelia were transferred onto poly -L -lysine treated cover slides and cultured overnight .Gene transfer was performed by elec‐troporation in medium containing pGPHI/GFP/Neo plasmid or by incubation with diluted recombined adenovirus/lentivirus vector .After 48 hours ,green fluorescence was checked under fluorescence microscope .To confirm the ef‐ficiency of exogenous gene transfer ,real-time PCR was performed using specific primers .Results The transfec‐tion efficiencies of electroporation and lentivirus vector mediated gene transfer were very low .Both immunofluores‐cence and real - time PCR results showed that the transfection efficiency of adenovirus mediated GFP and Bmi1 transfer were relatively higher .The proportion of GFP positive cells in outer hair cells and inner hair cells of middle turn were 90 .0% ± 4 .1% and 5% ± 0 .4% ,respectively .Conclusion Adenovector is more efficient for exogenous gene transfer to cochlear hair cells ,thus adenovector is a good carrier for gene transfer to cochlear hair cells .

Hypoxia occurs in chronic and acute vascular diseases and tumor formation. The ability of tumor cells to maintain a balance between an adaptation to hypoxia and cell death is regulated by a family of transcription factors called hypoxia-inducible factor 1 (HIF-1). Tumor hypoxia mediated by HIF-1 would facilitate the likelihood of resistance to chemotherapy and radiotherapy, proliferation, metastasis and the invasive potential; all of which culminate in a decrease in patient survival. And HIF-1 alpha subunit decides the activity of HIF-1, which is regulated by oxygen. So understanding the role of HIF in signal pathway, drug resistance mechanism and its feature is crucial for developing novel anticancer therapies. In recent years, more attentions have focused on HIF-1 alpha inhibitors. It is expected that development of more potent and selective HIF inhibitors will provide an effective treatment of cancer and other HIF-related diseases. So we will focus on the biological characteristics and mechanism of HIF-1 to review currently studied HIF-1 inhibitors.

Hypoxia occurs in chronic and acute vascular diseases and tumor formation. The ability of tumor cells to maintain a balance between an adaptation to hypoxia and cell death is regulated by a family of transcription factors called hypoxia-inducible factor 1 (HIF-1). Tumor hypoxia mediated by HIF-1 would facilitate the likelihood of resistance to chemotherapy and radiotherapy, proliferation, metastasis and the invasive potential; all of which culminate in a decrease in patient survival. And HIF-1 alpha subunit decides the activity of HIF-1, which is regulated by oxygen. So understanding the role of HIF in signal pathway, drug resistance mechanism and its feature is crucial for developing novel anticancer therapies. In recent years, more attentions have focused on HIF-1 alpha inhibitors. It is expected that development of more potent and selective HIF inhibitors will provide an effective treatment of cancer and other HIF-related diseases. So we will focus on the biological characteristics and mechanism of HIF-1 to review currently studied HIF-1 inhibitors.
Cell Death ; Humans ; Hypoxia-Inducible Factor 1 ; antagonists & inhibitors ; metabolism ; Hypoxia-Inducible Factor 1, alpha Subunit ; antagonists & inhibitors ; metabolism ; Neoplasms ; drug therapy ; Oxygen ; metabolism ; Signal Transduction

Objective: Immunotoxin rRTA:DS27, which was prepared by conjugating DS27 with recombinanl ricin a chain, was compared with ricin:DS27 as immunotoxins. Methods: System analysis were performed regard to their cell-specific cytotoxicity, inhibition on the proliferation of hemoapeutic potential cells, immunogold-labelled intracellular routing and effect of NH_(4)Cl on the cytotoxicity. Results: Results showed that rRTA: DS27 got a more specific cytotoxicity and a weaker inhibition on the proliferation of hemoapeutic potential cells than ricin:DS27, NH_(4)Cl could obsolutelyy enhence the cytotoxicity of rRTA:DS27. Conclusion: rRTA:DS27 had more advantages than ricin: DS27 as immunotoxins.

Objective:To build the model of the gene FKBP38 (FK506 binding protein 38) conditional knock out in liver.Methods:Transgenic mouse whose FKBP38 gene was flanked with loxP was constructed by embryo microinjection.The FKBP38 gene was deleted by breeding mice harboring two loxP sites in FKBP38 (FKBP38fl/fl) with the mice bearing the expression ofCre recombinase mice driven by an album promoter.Afterward,the genotype of FKBP38 conditional knockout mice was analyzed.Results:①Relative hepatic FKBP38 mRNA levels showed significant difference between FKBP38 conditional knockout mice (FKBP38-/-) and wild type(P＜ 0.001).②Relative hepatic FKBP38 protein expression levels of FKBP38 conditional knockout mice (FKBP38-/-) were significantly different with wild type(P＜0.001).③Relative phosphorylation of hepatic p70 S6K and 4E-BP-1 protein of FKBP38 conditional knockout mice (FKBP38-/-) showed no significant difference,with slight decrease in phosphorylation of 4E-BP-1,compared with wild type.④No significant difference in expression of hepatic Bcl-2 between FKBP38-/-and wild type.Conclusions:The mouse model of the gene FKBP38 (FK506 binding protein 38) conditional knock out in liver is successfully built.

AIM: To examine the microsatellite instability (MSI) and loss of heterozygosity (LOH) in head and neck squamous cell carcinomas (HNSCC). METHODS: 36 cases of HNSCC were analyzed with 15 microsatellite markers from chromosome 3,5,6,8,9,13,17 and 18. RESULTS: Among the 36 cases of HNSCC, 27.8%(10/36)of samples showed MSI in one to eight microsatellite markers. High frequent MSI occurred at D17S520(22.9%), D6S105(16 7%)and D8S264(13 9%). LOH was detected on the site of 9p21-p22 and 3p14. No correlations were found between allelic instability and grade or stage of the tumor. CONCLUSION: Our data suggest that MSI is a common genetic change in HNSCC. Tumor suppressor genes related to HNSCC may harbor at chromosome 9p21-p22 and 3p14 regions. [

Objective To investigate the needs of the main caregivers of children with cerebral palsy and probe into the nursing strategies. Method One hundred and fifteen main caregivers of children with cerebral palsy participated in the survey by a general state questionnaire and the family needs questionnaire. Results The score by the family needs questionnaire was (96.13 ± 19.47), with an average score of (3.00 ± 0.61). And the top needs of the main caregivers were information and econony need. Conclusions The needs of the main caregivers of children with cerebral palsy are high. More attention should be paid to community and family-based rehabilitation and supports to those care-givers.

ObjectiveTo evaluate the iodine nutritional status of university students in Tianjin and analyze influencing factors affecting urinary iodine levels.MethodsStudents of Tianjin Medical University,Tianjin Nankai University,Tianjin University of Finance and Economics and Tianjin University of Traditional Chinese Medicine were selected as survey subjects,and 50 - 100 morning urinary samples were collected from each university,respectively.Urinary iodine was measured by arsenic-cerium catalytic spectrophotometry.The students were surveyed with questionnaires,which included family information,age, sex, specialty, iodine nutrition knowledge,source of drinking water,smoking or not and dietary habits.ResultsA total of 269 urine samples were collected,and the median urinary iodine was 213.68 μg/L.Urinary iodine levels(263.86 μg/L) of medical students was significantly higher than that( 168.01 μg/L,x2 =12.144,P ＜ 0.01 ) of non-medical students.There was an increasing trend of the level of urinary iodine of students with iodine nutrition knowledge scores ＞ 5 points (223.70 μg/L) over that of ≤5 points( 185.56 μg/L),but the difference was not significantly different statistically (x2 =2.297,P ＞ 0.05).Different gender and water sources had no significant effect on urinary iodine level(x2 =0.002,0.687,respectively,all P ＞ 0.05).Smokers urinary iodine levels( 154.55 μg/L) decreased compared with non-smokers(215.38 μg/L),but the difference was not statistically significant (x2 =0.515,P＞ 0.05).Vegetarian urinary iodine levels were lower than that of non-vegetarians,but the difference was not statistically significant(x2 =0.594,P ＞ 0.05).ConclusionsIodine nutritional status of students in university of Tianjin are generally at an appropriate level,but professional knowledge,habits and other factors may affect the intake of iodine,so students should develop good dietary habits to ensure a normal iodine nutrition status.

To investigate the effect of compound Chinese traditional medicine PC-SPES II I in inhibiting proliferation of human prostate cancer cell LNCaP based on the androgen receptor (AR) signaling pathway. The effect of PC-SPES II on LNCaP cell proliferation was detected by MTT assay. According to the findings, at the mass concentration of 180-1 440 mg x L(-1), PC-SPES II significantly inhibited the proliferation of LNCaP cells; the IC50 of PC-SPES II at 24 h and 48 h were 311.48, 199.01 mg x L(-1), respectively. The flow Cytometry detection showed 240 mg x L(-1) PC-SPES II arrested cells in G2/M phase, and an obvious apoptotic peak appeared before G0/G1 peak and rose over time. Meanwhile, Hoechst 33258 staining revealed apoptotic cellular morphology. Annexin V-FITC/PI staining manifested an increase in apoptotic cell ratio at the PC-SPES II concentration of 480 mg x L(-1) in a dose dependent manner. The prostate specific antigen (PSA) secretion of LNCaP cells was tested by PSA ELISA kit. Besides, compared with 25 mg x L(-1) Bic, 480 mg x L(-1) PC-SPES II significantly reduced the cell secretion of PSA. The AR and PSA mRNA and protein expressions were detected by qRT-PCR and Western blot. According to the results, after the induction of LNCaP cells with synthetic androgen 25 μg x L(-1) R1881, 240-480 mg x L(-1) PC-SPES II notably down-regulated the AR and PSA mRNA and protein expressions and inhibited the translocation of AR from cytoplasm to nucleus. In summary, PC-SPES II significantly can inhibit the in vitro proliferation of LNCaP cells and arrest cell cycle arrest in G2/M phase. Its mechanism may be associated with the down-regulation of the AR and PSA expressions and the inhibition of AR nuclear translocation.
Antineoplastic Agents, Phytogenic ; pharmacology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; Humans ; Male ; Prostate-Specific Antigen ; genetics ; metabolism ; Prostatic Neoplasms ; drug therapy ; genetics ; metabolism ; physiopathology ; Receptors, Androgen ; genetics ; metabolism ; Signal Transduction ; drug effects

Objective To study the mechanism of SW480 cell line death caused by transfection of retroviral vector-G1CEACDNa.Methods Plasmid G1CEACDNa was transferred into the SW480 cell line using liposomes method. RT-PCR was performed to examin the expression of CD gene indrectly.The cell growth curve was measured by means of cell counting. When the CD+SW480 cells were exposed to 5-FC (1mmol/L), the growth inhibition rate and the "bystander effect" were detected by MTT method.The ultrastructure was observed by electron microscope.Apoptosis was verified by flow cytometer .Results The product of RT-PCR showed a band at 1.5kb on the photo of electrophoresis. The growth of CD+ SW480 cells was inhibited 24h after administrating 5-FC,and the inhibition rates at 72h,96h,120h were 30.0%,50.0% and 80.0%,respectively.Apoptosis cells in different phases and apoptotic bodies in the field of electron microscope were observed. Meanwhile ,a few cells showed necrosis.Flow cytometer verified that a few cells appearred apoptosis 48h after exposed to 5-FC (1mmol/L), the apoptosis rate and the necrosis rate at 72h,96h were 20.2%,30.7% and 19.6%,21.1% respectively.Conclusions The death mechanism of SW480 cells transfected with G1CEACDNa followed by 5-FC treatment includes both necrosis and apoptosis.Apoptosis is possibly the bystander effect.