Objective To construct the yeast expression vector of a new gene transactivated by hepatitis B virus X protein (XTP11), and to explore the feasibility of cloning the hepatocyte proteins interacting with XTP11 protein by yeast two hybrid system. Methods The XTP11 gene was amplified by polymerase chain reaction (PCR) with specific primers, and the amplified fragment was subcloned into the Nco I/BamH I sites (5′ ends) of yeast expression vector pGBKT7,which was named as pGBKT7(-)-XTP11 encoding fusion proteins of full length of XTP11 and DNA binding domain of yeast protein GAL4. The pGBKT7 (-)-XTP11 plasmid was transformed in the yeast cells AH109 and the expression of XTP11 protein in yeast cells was detected by Western blotting assay. Then the yeast cells were plated on synthetic dropout nutrient medium (SD/-Trp and SD/-Trp-His-Ade) containing X-?-gal, and their autonomous activation was tested. Results The yeast expression vector of XTP11 gene was constructed and the fusion protein in yeast cells was examined. The yeast cells transformed with pGBKT7-XTP11 plasmid could grew well on both of the media, and turned blue on medium containing X-?-gal for the production X-?-galactosidase. This phenomenon suggested that XTP11 protein acted to substitute the functional yeast protein GAL4 activation domain (AD) to activate transcription of reporter genes (ADE2, HIS3, MEL1 and LacZ). Conclusion The XTP11 protein fused to the GAL4 DNA-binding domain functioned as a transcriptional activation domain in yeast, and the transcriptional activation function of XTP11 protein in yeast might restrain researchers to use yeast two hybrid system to clone hepatocyte proteins interacting with XTP11 protein.

BACKGROUND: Anterior decompression and internal fixation for treatment of cervical spondylotic myelopathy has been widely accepted. However, there are few clinical case reports regarding this.OBJECTIVE: To investigate the curative efficacy of anterior cervical decompression combined with bone grafting and titanium plate internal fixation in treatment of cervical spondylotic myelopathy.METHODS: A total of 118 patients with cervical spondylotic myelopathy who underwent anterior cervical subtotal vertebrectomy,iliac bone grafting (or bone grafting with titanium cage) and anterior locking titanium plate internal fixation at the Department of Orthopedics, Huang Gang Central Hospital of Hubei Province between January 2001 and August 2007 were retrospectively analyzed. According to the X-ray photographs taken before, immediately after surgery and during postoperative follow-up periods,Cobb's angle, anterior vertebral height and posterior vertebral height of fused segments. Bone graft fusion was evaluated.Neurological function was assessed according to Japanese Orthopaedic Association (JOA) scores.RESULTS: During the follow-up period (6-32 months, mean 19 months), three patients presented with slight titanium mesh sinking, and no fragmentation or loosening during internal fixation was observed in all patients, with a final bone fusion rate of 100％. JOA scores obtained immediately after surgery and during last follow-up were significantly greater compared with those obtained prior to surgery (P ＜ 0.05). The excellent and good rate of neurological function recovery during the last follow-up was 86.2％. After surgery, sagittal Cobb's angle, anterior and posterior vertebral heights of fused segments were significantly improved(P ＜ 0.05). These findings indicate that anterior cervical subtotal decompression combined with bone grafting and titanium plateinternal fixation for treatment of cervical spondylotic myelopathy can thoroughly decompress, effectively correct cervical vertebrae malformation, and achieve stable bone fusion and reconstruction, with satisfactory clinical efficacy.

BACKGROUND:Lumbar fusion surgery as an important and effective means of treating degenerative diseases is widely used in clinical application for almost a century. However, long-term clinical evidence showed that lumbar fusion also brought some problems, such as the loss of waist flexibility, complications of donor site, fusion segment motion loss and accelerating the adjacent segment degeneration. In recent years, the theory of spinal dynamic stabilization had spread widely, and a variety of non-fusion surgery is becoming more broadly used in treatment of lumbar degenerative disease. OBJECTIVE:To review the application of non-fusion surgery in the treatment of lumbar degenerative diseases and explain application perspectives and experiences. METHODS:Databases including PubMed and CNKI were retrieved to col ect clinical application and views about non-fusion internal fixation in the treatment of lumbar degenerative diseases from 2007 to 2016. The key words were“lumbar, non-fusion, bone fusion, dynamic stabilization, adjacent segment degeneration”. RESULTS AND CONCLUSION:Thirty papers were included in the final analysis after screening by two independent researchers. We summarized the lumbar non-fusion technology, including artificial nucleus replacement, total disc replacement and posterior lumbar dynamic stabilization device. The philosophy of these new technologies is to provide stability and physiological activity, reducing the abnormal stress that leads to adjacent segment degeneration. These methods can be applied to the step of treatment of lumbar degenerative diseases and to reduce the fusion of diseased segments. Simultaneously, it is needed to strictly grasp the indications for surgery, to identify the cause of the pain caused by low back pain and lumbar spine instability, and to select the most suitable non-fusion device for individual treatment.

Hepatitis B virus (HBV)covalently closed circular DNA (cccDNA)is an intermediate replicative episome which is stable and not easy to eliminate.cccDNA serves as the template for HBV replication and plays an important role in persistence of HBV infection in hep-atocytes,and it is also a biomarker of HBV activity.Studies suggested that HBV has extrahepatic infectivity and cccDNA could be detected in peripheral blood mononuclear cells (PBMC).The predictive values of HBV cccDNA in PBMC for HBV recurrence after liver transplanta-tion and antiviral therapy and mother-to -child transmission of HBV in patients with hepatitis B,as well as the possible mechanism of cccDNA involved in extrahepatic HBV infection,are summarized.Therefore,the detection of HBV cccDNA is of great significance for the study and treatment of HBV.

Objective:To explore the possible role and clinical significance of Th1 and Th2 class cytokines in the pathogenesis of re-current aphthous ulcer (RAU).Methods:35 cases with RAU,25 of minor RAU(MiRAU)and 1 0 of major RAU(MaRAU),were enrolled and 24 healthy subjects served as the controls.The expression levels of Th1 type cytokines (IFN-γ,TNF-α,IL-2)and Th2 type cytokines (IL-4,IL-6,IL-1 0)in the serum of peripheral blood were investigated by flow cytometry CBA methods and their rela-tionship with the clinical features of RAU was analyzed by SPSS 1 1 .5.Results:The expression level of IL-4 and IL-1 0 in MiRAU group was higher than that in the controls(P <0.05),and the expression level of IFN-γ,IL-4 and IL-1 0 in MaRAU group was higher than that in the controls(P <0.05).The expression level of IFN-γand IL-1 0 in MaRAU was higher than that in MiRAU.There was a positive correlation between the levels of IFN-γ,IL-4,IL-1 0 and the severity of RAU.Conclusion:Predominant Th2 immune re-sponse plays a role in RAU,and the level of IFN-γ,IL-4 and IL-1 0 is correlated with the severity of RAU.

Objective To observe the effect of Yiqi Fuzhi Granule on activating blood circulation and removing blood stasis in rat models of multiple infarctional dementia.Methods Internal carotid artery injection method was used to establish rat models of multiple infarctional dementia.The changes of blood rheology,blood platelet aggregation,thromboplastin time,plasmozyme time and kaolin partial thromboplastin time before and after operation were observed.Results Yiqi Fuzhi Granule could markedly improve blood rheology in rat models of multiple infarctional dementia and reduce blood viscosity and blood platelet aggregati on rate,the effect being obvious before operation.Conclusion Yiqi Fuzhi Granule has a good action of activating blood circulation and removing blood stasis.

This paper reports the efficacy of the treatment of local swelling caused by intravenous injection Kakage with hot (40-50℃) or cold (4-6℃) compress. It was found that cold compress had a better repellent effect than hot one. Cold compress could resolve the swelling due to extravasation rapidly, effectively and safely.

Objective To discuss the value of Valsalva experiment combined with improved MRI tilt tomography applying on the pharyngeal ostium of eustachian tube opening function of patients with nasopharyngeal carcinoma.Methods 1 7 cases with patholog-ically proved NPC underwent conventional MRI plain and enhanced scan.In the eustachian tube area oblique transection T2 WI and T1 WI scanning was done before and after the Valsalva experiment.The invasion of eustachian tube and its surrounding structures was analyzed and the eustachian tube function was evaluated.Results Valsalva experiments combined with improved MRI trans-verse oblique scan could clearly display the whole structure of the eustachian tube and surrounding tissue invasion compared with conventional scanning MR scan.And it could better evaluate the eustachian tube opening function for patients suffered nasopharynge-al carcinoma.Conclusion Valsalva experiments with improved MRI oblique transverse imaging could achieve individualized scan of the eustachian tube,and could be used for evaluating the pharyngeal opening function of eustachian tube in patients with nasopharyn-geal carcinoma.

To better understand the cleavage efficiency of muhiribozyme system on its RNA substrate in the presence and absence of divalent magnesium and monovalent sodium ions.we constructed pGEM-Coat'A,pGEM-Coat'A196Rz plasmids and pGEM-MDRl target plasmid.They were applied to transcribe RNAs with SP6/T7 transcription kit.Cleavage reactions were carried out in cell-free system and reaction products were analyzed by electrophoresis on 6% denaturing polyacrylamide gels in TBS buffer.The gels were dried and exposed to X-ray films for autoradiography.The Image J software was employed to analyze the dried gels.The results indicated that the cleavage efficiency of the muhiribozyme was dependent on the concentration of divalent Mg2+.The cleavage products increased with the concentrations of divalent Mg2+ and were Mg2+ concentration and time dependent.No cleavage product was obtained in the presence of lower than 200 mmol/L Na+ alone.On the contrary,monovalent Na+ inhibited the Mg2+ -induced cleavage reaction in Na+ and Mg2+ coexistance.The cleavage rate was significantly lower than that observed with divalent Mg2+ alone.These results suggested that divalent Mg2+ was required for muhiribozyme on substrate cleavage reaction in the physical condition,whereas monovalent Na+ was not.

10.3969/j.issn.2095-4344.2013.23.002