Objectives: To assess the feasibility and stability of right ventricular outflow tract (ROVT) pacing under current technology by comparing the results of ROVT pacing with the traditional right ventricular apex (RVA) pacing. Methods: A total of 42 patients (at mean age of 63.5±10.4 years) without structural heart disease were randomly divided into two groups. RVA pacing group (n=14),and RVOT pacing group(n=28). An active fixation lead was implanted in all patients whose pacemaker could automatically measure the pacing threshold every day. The operation time,X-ray exposure time and lead parameters detected during the operation were collected to evaluate the feasibility of RVOT pacing. The complications related to lead and implantation procedure and the trend of threshold change during the follow-up time were used to assess the stability of RVOT pacing.Results: There were no statistic differences between RVA pacing group and RVOT pacing group in terms of operation time,X-ray exposure time and lead parameters. In RVOT group,the change of threshold during acute period was similar to those in RVA group (P=0.23). Chronic pacing threshold was also comparable between two groups,mean threshold at 6 months follow-up time was 0.55±0.11V and 0.54±0.09V at 0.4 pulse width in RVA group and RVOT group respectively (P=0.787).Conclusion: RVOT pacing was feasible and stable in operation time and lead characteristics compared with the conventional RVA pacing under current pacing technology.

BACKGROUND: Central nervous system (CNS) myelin is made up of 70% lipids and 30% proteins with human brain myelin basic protein (hMBP) constituting 1/3 of the proteins. MBP is a family of proteinswith four isoforms only in human brain. OBJECTIVE: To investigate the expression of MBP and its immunological function. DESIGN: Single sample study. SETTING: Department of Biochemistry and Molecule Biology, Huaxi College of Priclinical Medicine and Forensic Medicine, Sichuan University. MATERIALS: This experiment was conducted at Department of Bio chemistry and Molecule Biology, Huaxi College of Priclinical Medicine and Forensic Medicine, Sichuan University from August 2003 to March 2004. Relative molecular weight was 215000 hMBP cDNA clone pGEMP,prokaryotic expression recombinant vector pGEX-5T, T4 DNA Ligase,calf intestine alkaline phosphates, X-gal, IPTG, 123 Ladder (BRL), nitrocellu lose ,4-chloro-1-Naphthol;MBP and Anti- MBP ELISA kit. INTERVENTIONS: ①hBMP gene cDNA clone segment was digested with EcoR1 and BamH1; ②Construction and transformation of the recombinant expression vector p5TMP; ③ The growth and induced expression of the recombinant expression vector transformed bacteria; ④ Preparation of the antibody of recombinant hMBP. MAIN OUTCOME MEASURES: ① Detection and identification of the expressed protein product; ② Detection and identification of hMBP anti body. RESULTS: ① Screening and identification of recombinant MBP: The 4 999 bp pGEX-5T DNA and 557 bp MBP inserted fragment were obtained, indicating that the white colonies contained recombinant expression plasmid of exogenous DNA; ② A dense band disappeared from the control plasmid while a new special polypeptide band with apparent molecular weight 42 000 was detected in recombinant cell lysate; ③After 5 subcutaneous injections, antibody was obtained at 1:16 titer. The specificity of the antibody to MBP was confirmed.CONCLUSION: Compared with the original expression vector, the new constructed expression vector containing 215 000 MBP exons Ⅰ-Ⅶ coding sequence was not only without the coding area defects of 34 bp in 5' sequence but also expressed more highly in prokaryote obviously. In addition, the recombinant MBP antibody prepared successfully.

We constructed the expression vector by inserting 21.5 KDa MBP human brain full-length cDNA coding sequence digested with restriction enzyme EcoR I and Sal I into downstream of pGEX-5T expression vector. The recombinant vector p5TMP was transformed into E. coli and the positive clonies were selected and incubated in LB medium induced by IPTG (isopropyl- -D-thiogalactoside). A new polypeptide band with apparent molecular weight 42 KDa was detected in transformed cell lysates by SDS-PAGE. Western blotting analysis confirmed that this fusion protein reacted specifically with antibodies to MBP, the expression level of MBP was about 414.6 mg/L medium estimated by immuno-dot blot, ELISA and absorbance scanning. Newzealand rabbits were immunized by subcutaneous injection of the purified recombinant MBP. The titer was obtained at 1:16 after 5 injections. The specificity of the antibody to MBP was confirmed by immuno-blot and Western blotting.
Animals ; Antibody Formation ; Autoantibodies ; Brain ; immunology ; metabolism ; Cloning, Molecular ; DNA, Complementary ; Escherichia coli ; genetics ; metabolism ; Gene Expression ; Humans ; Myelin Basic Protein ; biosynthesis ; genetics ; immunology ; Rabbits ; Recombinant Fusion Proteins ; biosynthesis ; immunology

OBJECTIVEThe purposes of this study were to clone and sequence the major histocompatibility complex type I (MHC I) molecular antigen recognizing gene (H-2Kk) of 615 mice, and to provide the functional gene for transgenic therapy.

METHODSThe 1.4 kb full-length fragment of H-2Kk gene complementary DNA (cDNA) was amplified from the total RNA of 615 mouse liver by using reverse transcription polymerase chain reaction (RT-PCR). The cDNA was inserted into PGEM3Zf(+) vector directionally, and the competent E. coli JM109 was transformed with the ligated product. The recombinant PGEM3Zf(+)-H-2Kk cDNA plasmid was obtained using restricted enzyme analysis of the transfectants. The complete sequence of 615 mouse H-2Kk cDNA was determined by using Sanger's method.

RESULTSThe sequences of 615 mouse H-2Kk cDNA were 99% similar with those of H-2Kk cDNA which were reported by other researchers, and the sequences encoding antigen recognizing regions (ARS) were identical with each other.

CONCLUSIONThe authors cloned the MHC I molecular antigen recognizing gene (H-2Kk) of 615 mice successfully and got the functional gene of MHC I.

Animals ; Cloning, Molecular ; DNA, Complementary ; genetics ; Escherichia coli ; genetics ; Genes ; genetics ; Genes, MHC Class I ; genetics ; Genetic Therapy ; H-2 Antigens ; genetics ; Mice ; Mice, Inbred C57BL ; Point Mutation ; Sequence Analysis, DNA ; Transgenes

The primers specific for the full-length BDNF coding sequence was designed and synthesized. The BDNF coding sequence was directly amplified from human genomic DNA by using PCR and inserted into vector pGEM-3Zf(+). The recombinant DNA was transformed into the host cells JM109 to obtain the positive clone pGEMBF18. The restriction enzyme analysis and DNA sequence detection confirmed that the inser ted fragment of clone pGEMBF18 is the full-length BDNF coding sequence. The hBD NF DNA fragment was recovered from the clone pGEMBF18 and ligated with prokaryot ic expression vector pGEX-5T to construct the recombinant expression plasmid p5 TBF34. The E.coli JM109 transformed with p5TBF34 was induced with IPTG. A new pr otein band with apparent molecular weight 43 kDa was detected in the lysate of t he transformed cell by using SDS-PAGE. The result of western hybridization show ed that this fusion protein reacted specifically to the antibodies to human BDNF . The amount of the soluble fusion protein was about 503.04mg/L lysate, 7.53% of total bacterial soluble protein of transformed cells, estimated by absorbance sc anning of SDS-PAGE and protein quantitation.

Humans ; Cardiac Resynchronization Therapy ; Cardiac Pacing, Artificial ; Heart Ventricles ; Heart Failure/therapy* ; Treatment Outcome ; Electrocardiography ; Ventricular Function, Left

Objective To screen the pathogenic gene of osteopetrosis to provide reference for its genetic di-agnosis and prognosis .Methods The clinical data and peripheral blood samples were collected from the pa-tients with osteopetrosis ,DNA was extracted ,the whole exome sequencing library was built ,then the high throughput detection was performed and the pathogenic gene was screened by combining with the bioinformat-ics technology .Results The whole exomes in 2 cases of osteopetrosis were analyzed ,the average sequencing depth of the two samples were 169 .38X and 231 .06X respectively ,in which the case 1 carried rare mutation TCIRG1(c .1305+2T>C) ,TCIRG1(c .2008C> T ) and CLCN7(c .1116C> T );the case 2 carried a rare muta-tion CLCN7(c .857G>A ) .T he bioinformatics analysis indicated that the rare mutations carried by these cases all had different degrees of influence on the structure and function of gene products .Conclusion The whole exome sequencing can once screen the know n pathogenic mutations of osteopetrosis ,is an effective tool for pathogenic mutation screening of osteopetrosis ,the clinical disease in 2 cases of osteopetrosis may be closely related with the patient′s carrying TCIRG1 and CLCN7 mutation .

This study explored the ideas and methods of acupuncture for Guillain-Barré Syndrome （GBS） with the core principle of “to treat flaccidity， select the yangming （阳明） channel only”. The main pathological mechanism of GBS is deficiency of qi and blood in the yangming channel， malnutrition of all sinews， diminished spleen and stomach function leading to the production of pathogenic damp-heat qi， which obstructs the meridians， and gradually affects the liver and kidneys， consuming essence and damaging blood. Concurrently， dysfunction of the dumai （督脉） pivotal mechanism and lack of moisture in sinews and vessels result in symptoms such as skin numbness， paralysis， and muscle wastage. In clinical diagnosis and treatment， a combination of syndrome and channel differentiation is taken. Treatment primarily focuses on acupoints of yangming channel， aiming to supplement qi and blood， and acupoints of du mai are combined to open the vessel and fill the marrow. Specific acupoints are selected based on syndrome differentiation， providing comprehensive regulation to promote harmonization of qi and blood， relieve meridians， and the smooth generation and circulation of whole body fluids. This， in turn， enhances the strength of muscles and bones， and fosters a robust and freely moving body.