Objective To investigate Glypican-3 gene expression and mutation in hepatocellular carcinoma (HCC).Methods Glypican-3 gene expression and mutation in tumor,para-c.ancer and normal tissue of 48 HCCs were detected by RT-PCR and single-strand conformation polymorphism(SSCP),respectively.Results There was no Glypican-3 mRNA expression in para-cancer and normal tissue.Expression rate of Glypican-3 mRNA was 77.1% in tumor tissue,which was correlated with clinical staging and cell differentiation(P

OBJECTIVETo evaluate the role of p53 gene mutation in colorectal carcinoma and assess the value of peripheral blood p53 single nucleotide polymorphism (SNP) in early diagnosis of colorectal carcinoma.

METHODSNSP in axons 5-8 of p53 gene was detected using ligase detection reaction-polymerase chain reaction (LDR-PCR) in the peripheral blood of 100 patients with colorectal cancer and 100 healthy subjects.

RESULTSThe mutation rate of p53 gene was 24% (24/120) in colorectal carcinoma patients and 0% (0/120) in the healthy subjects (P<0.05).

CONCLUSIONp53 plays an important role in the carcinogenesis of colorectal carcinoma, and SNP analysis for p53 gene can be helpful in early diagnosis of colorectal carcinoma.

Adult ; Case-Control Studies ; Colorectal Neoplasms ; diagnosis ; genetics ; DNA Mutational Analysis ; DNA, Neoplasm ; genetics ; Early Detection of Cancer ; Female ; Humans ; Male ; Mutation ; Polymorphism, Single Nucleotide ; Tumor Suppressor Protein p53 ; genetics

Objective To make a systematic review of therapeutic effect of electroacupuncture(EA) for idiopathic facial palsy at acute stage. Methods With reference to the included criteria and excluded criteria, clinical controlled trials of EA for idiopathic facial palsy at acute stage were screened from the databases of CBM, CNKI, Wanfang, VIP by computer retrieval and from the primary domestic academic journals of acupuncture by manual retrieval. Systematic review was performed following the Cochrane Reviewers' Handbook, and RevMan 5.3 was used for Meta-analysis. Results Sixteen clinical trials were included. The Meta-analysis results showed that the cure rate evaluated with 3 kinds of therapeutic effect criteria in the observation group was superior to that of the control group, the difference being significant (P<0.05 or P<0.01); the effects on improving days for cure, cure case number within one month, blink reflex, and case number with complications in the observation group were also superior to that of the control group, the difference being significant (P < 0.01). Conclusion EA exerts certain therapeutic effect for idiopathic facial palsy at acute stage. But for the quantity and quality of the included trials are not satisfactory, the conclusion still needs more high-quality, large-size and long-term follow-up trials to verify.

The aim of study was to investigate the killing effect of double suicide gene system mediated by retroviral vector on K562 cells in vivo and ex vivo. CDglyTK gene was transfected into PA317 cells by using lipofectamine. K562 cells were infected with viral supernatant. K562/CDglyTK cells were treated with 5-fluorocytosine (5-FC) and/or ganciclovir (GCV). Mice were randomly divided into three groups: tumor formation, tumor inhibition and tumor therapy. Each mouse was implanted with K562/CDglyTK cells or K562 cells. The results indicated that the killing effect of 5-FC in combination with GCV on K562/CDglyTK was more significant than using 5-FC or GCV alone. In vivo study showed that after being injected subcutaneously with K562 cells and K562/CDglyTK cells, there was not obvious difference in tumor formation rate of mice, 5-FC + GCV could suppress tumor formation of the K562/CDglyTK cells. After being treated with 5-FC and GCV, the median tumor volume of mice implanted with K562/CDglyTK cells decreased obviously, compared with the control group. Their median survival was significantly prolonged. It is concluded that double suicide genes are more effective for killing effect on K562 cells in vivo and in ex vivo. It may be applicable to clinical gene therapy.
Cytosine Deaminase ; genetics ; Flucytosine ; pharmacology ; Ganciclovir ; pharmacology ; Genes, Transgenic, Suicide ; genetics ; Genetic Therapy ; Genetic Vectors ; genetics ; Humans ; K562 Cells ; Protein-Tyrosine Kinases ; genetics ; Receptor Protein-Tyrosine Kinases ; biosynthesis ; genetics ; Recombinant Fusion Proteins ; genetics ; Recombination, Genetic ; Retroviridae ; genetics

To investigate the expression and localization of various functional domains of ORF1 polyprotein and ORF3 protein of hepatitis E virus in host cells, the coding sequences of the various functional domains (RdRp, HEL, MET, PLP, X) of ORF1 were separately cloned into pcDNA3. 1-GFP vectors for constructing the recombinant plasmids which were verified by enzyme digestion and sequencing. The exact expression of the fusion proteins were detected by Western Blot, and the distribution and localization were observed by the laser scanning confocal microscope(LSCM). In huh7 cells, GFP-RdRp proteins were found mainly in the nuclei, GFP-HEL proteins were distributed vesicularly around the nucleus, GFP-MET proteins were distributed granularly both in the nuclei and the cytoplasm, GFP-PLP proteins had polar distribution around the nucleus, and unknown GFP-X proteins were distributed uniformly both in the nuclei and the cytoplasm. Different localization of these proteins verified the previous data obtained from in vitro studies, providing a support for further research on the biological functions of various proteins coded by HEV genome.
Blotting, Western ; Cells, Cultured ; Hepatitis E virus ; genetics ; Humans ; Open Reading Frames ; Viral Proteins ; genetics ; physiology

Adult ; Female ; Humans ; Kidney Neoplasms ; diagnostic imaging ; metabolism ; Neuroectodermal Tumors ; diagnostic imaging ; metabolism ; Radiography ; Young Adult

OBJECTIVETo investigate the effect of M(5) muscarinic receptor subtype on the locomotor sensitization induced by heroin priming, and it's effect on the FosB expression in the nucleus accumbens (NAc) and the hippocampus in the heroin sensitized rats.

METHODSLocomotor activity was measured every 10 min for 1 h after subcutaneous injection of heroin. FosB expression was assayed by immunohistochemistry, and the antisense oligonucleotides (AS-ONs) targeting M(5) muscarinic receptor was transferred with the lipofectin.

RESULTSMicroinjection of AS-ONs targeting M(5) muscarinic receptor in the ventral tegmental area (VTA) blocked the expression of behavioral sensitization induced by heroin priming in rats. Meanwhile, the expression of FosB-positive neurons in either the NAc or the dentate gyrus (DG) of the hippocampus increased in heroin-induced locomotor sensitized rats. The enhancement of FosB-positive neurons in the NAc or DG could be inhibited by microinjection of M(5) muscarinic receptor AS-ONs into the VTA before the heroin-induced locomotor sensitization was performed. In contrast, microinjection of M(5) muscarinic receptor sense oligonucleotide (S-ONs) into the VTA did not block the expression of behavioral sensitization or the expression of FosB in the NAc or DG in the heroin sensitized rats.

CONCLUSIONBlocking M(5) muscarinic receptor in the VTA inhibits the expression of heroin-induced locomotor sensitization, which is associated with the regulation of FosB expression in the NAc and hippocampus neurons. M(5) muscarinic receptor may be a useful pharmacological target for the treatment of heroin addiction.

Acetylcholine ; metabolism ; Animals ; Brain ; drug effects ; metabolism ; physiopathology ; Heroin ; adverse effects ; Heroin Dependence ; drug therapy ; metabolism ; physiopathology ; Hippocampus ; drug effects ; metabolism ; Immunohistochemistry ; Male ; Microinjections ; Motor Activity ; drug effects ; physiology ; Narcotics ; adverse effects ; Neural Pathways ; drug effects ; metabolism ; physiopathology ; Neurons ; drug effects ; metabolism ; Nucleus Accumbens ; drug effects ; metabolism ; physiopathology ; Oligonucleotides, Antisense ; pharmacology ; Proto-Oncogene Proteins c-fos ; drug effects ; metabolism ; Rats ; Rats, Sprague-Dawley ; Receptor, Muscarinic M5 ; antagonists & inhibitors ; genetics ; metabolism ; Synaptic Transmission ; drug effects ; physiology ; Ventral Tegmental Area ; drug effects ; metabolism ; physiopathology

OBJECTIVETo assess the clinical usefulness, accuracy, and safety of tele-manipulation for frameless stereotactic surgery using the CAS-R-5 robot system.

METHODSWe prospectively evaluated 32 patients underwent tele-manipulation of frameless stereotactic operations from Sep. 2005 to Sep. 2006. Tele-manipulations were performed via a digital data network by a neurosurgeon in Beijing while the patients were located in Yan'an. The distance is 1300 kilometers away. The accuracy of location and improvement of symptom were observed after operation. The period of follow-up was from 3 to 14 months (the average was 12 months).

RESULTSThe surgical operations in 32 cases were successful. Remote fiducial registration was performed with a mean accuracy of 1. 50 mm and the standard difference were 0.32 mm between the planned and actual target. There were no complications.

CONCLUSIONSDiagnosis and treatment for intracranial disease by tele-manipulation frameless stereotactic surgeries are reliable and safe.

Adolescent ; Adult ; Aged ; Brain ; pathology ; surgery ; Brain Diseases ; surgery ; Child ; Child, Preschool ; Female ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Reproducibility of Results ; Retrospective Studies ; Robotics ; methods ; Stereotaxic Techniques ; Surgery, Computer-Assisted ; Treatment Outcome

Objective To analyze the effects of S100A6 on Wnt/β-catenin signaling pathway and its molecular mechanism. Methods The expression of GST-hS10OA6 was induced with IPTG in Escherichia coli BL21, and the fusion protein was purified with glutathione-sepharose 4B beads. β-catenin level of human colon cancer cell line MG63 and human osteosarcoma cell line HCTl16 cells infected with AdS10OA6 was measured by Western blot. Luciferase activity assay was applied to analyze the effect of S100A6 on the β-catenin/TCF4 activity. The interactions between S100A6 and β-catenin/GSK-3β/Dvl/Axin were detected by GST-pulldown/Western blot. Results The β-catenin level in AdS100A6-infected MG63 and HCT116 cells was significantly increased in comparison with that in the AdGFP control group (P<0.01). The luciferase activity in human embryonic renal cell line 293 cells transfected with pTOP-Luc and followed by GST-hS100A6 treatment was increased by 20. 2-fold in comparison with that in the GST control group (P<0.01). The interaction between GST-hS100A6 and Axin was not found. Conclusion S100A6 up- regulates the Wnt/β-catenin signaling pathway, and this may be attributed to the interaction between S100A6 and β-catenin/GSK-3β/Dvl.

Objective To analyze the effects of S100A6 on Wnt/β-catenin signaling pathway and its molecular mechanism. Methods The expression of GST-hS10OA6 was induced with IPTG in Escherichia coli BL21, and the fusion protein was purified with glutathione-sepharose 4B beads. β-catenin level of human colon cancer cell line MG63 and human osteosarcoma cell line HCTl16 cells infected with AdS10OA6 was measured by Western blot. Luciferase activity assay was applied to analyze the effect of S100A6 on the β-catenin/TCF4 activity. The interactions between S100A6 and β-catenin/GSK-3β/Dvl/Axin were detected by GST-pulldown/Western blot. Results The β-catenin level in AdS100A6-infected MG63 and HCT116 cells was significantly increased in comparison with that in the AdGFP control group (P<0.01). The luciferase activity in human embryonic renal cell line 293 cells transfected with pTOP-Luc and followed by GST-hS100A6 treatment was increased by 20. 2-fold in comparison with that in the GST control group (P<0.01). The interaction between GST-hS100A6 and Axin was not found. Conclusion S100A6 up- regulates the Wnt/β-catenin signaling pathway, and this may be attributed to the interaction between S100A6 and β-catenin/GSK-3β/Dvl.