The basic principle,characters and applications of I 2 C bus protocol are introduced in this paper.Its application to implantable e lectrical stimulator is also described based on MAX5630,a DAC chip produced by M AXIM Corporation.

Objective To develop a multiplex polymerase chain reaction (PCR )method for detecting and genotyping moxifloxacin-resistant Clostridium difficile (C.difficile)isolates.Methods Specific PCR primers of slpA genotypes gr,hr,fr,gc08 and 078 were designed according to the differences of slpA nucleotide sequences in different C.difficile genotypes,and the house-keeping gene tpi specific PCR primers were also added for the construction of multiplex PCR method.Nine common intestinal normal and pathogenic strains were used to verify the specificity of slpA multiplex PCR for the detection of C.difficile.Forty-six C.difficile reference strains,belonging to 11 slpA genotypes,were used to verify the ability of the multiplex PCR method for dectecting and genotyping.Thirty-nine moxifloxacin-resistant clinical isolates were genotyped by the multiplex PCR,and its clinical value was evaluated by comparing with slpA sequence typing (slpA ST)method.Results All the 9 intestinal normal and pathogenic strains were negative when detected by the multiplex PCR.And tpi of 46 C. difficile reference strains were positive,and 36 strains belonging to slpA genotypes gr,hr,fr,gc08 and 078 were genotyped correctly.Other 10 strains which belonged to other 6 genotypes were non-typeable. Among 39 moxifloxacin-resistant clinical isolates,all were positive of tpi,and 32 isolates were typed correctly by the multiplex PCR method,including 22 slpA genotypes gc08,6 genotypes hr,2 genotypes fr,and 2 genotypes 078,which were consistent with slpA ST.However,7 isolates could not be typed by multiplex PCR,which were identified as other genotypes not included in the multiplex PCR by slpA ST. Conclusions A convenient and rapid multiplex PCR method for the detection of C.difficile is established successfully,which can distinguish among five slpA genotypes.slpA genotype gc08 is the common genotype of moxifloxacin-resistant clinical isolates.

Ob jective To research the gene expression profile of aura-absence migraine patients before and after acupuncture of Shaoyang meridian acupoints or non-acupoints. Methods Twenty aura-absence migraine patients were randomly divided into meridian acupoint group and non-acupoint group, 10 cases in each group. Gene chip technology was used to investigate the differences of two sets of gene expression profiles, and reverse transcription-polymerase chain reaction (RT-PCR) was applied for the analysis of partial genes to verify the accuracy of gene chip detection results. Results Seventy-two differentially expressed genes were obtained in meridian acupoint group, and 110 differentially expressed genes were obtained in non-acupoint group. The function genes of meridian acupoint group involved brain endorphin enzyme, adenosine triphosphate ( ATP) synthase, etc., which were closely related with the curing of aura-absence migraine. Non-acupoint group had extensive and scattered function genes involving apoptosis, DNA repair, etc., which had less correlation with the curing of aura-absence migraine. ATPAF2, PTGS2, TOR3A genes of meridian acupoint group and ACP2, AURKA, ARHGEF11, CASP8 gene of non-acupoint group presented by RT-PCR analysis had verified the reliability of microarray data. Conclusion The therapeutic effect of meridian acupoints acupuncture for aura-absence migraine has achieved through the multi-gene action at the molecular level, but the corresponding target genes for the placebo effect of non-acupoint acupuncture have not been found , which demonstrates the existence of meridian effect.

Objective To study the expression and significance of acid-sensing ion channels(ASICs)in rat articular cartilage with adjuvant arthritis. Methods Complete Freund's adjuvant(CFA) was prepared by suspending heat-killed Bacillus Calmette Guerin(BCG) in liquid paraffin at 10 mg/ml. CFA-induced arthritis was developed by injection of 100 μl CFA emulsion intradermally into the right hind paw. The morphological changes of articular tissues was observed by light microscope; RT-PCR and immunoblotting analyses were used to detect ASICs in rat articular cartilage with adjuvant arthritis. Results RT-PCR and western blot showed that ASIC1a, ASIC2a and ASIC3 were present in the articular cartilage of normal and model group, the ASICs mRNA levels in the model group were higher than in the normal group detected by semiquantitative analysis (P＜0.01), ASICs protein levels in model group were higher than those in the normal group (P＜0.01) when examined by immunoblotting. Conclusion The results show that the expression of ASICs in AA articular cartilage is enhanced and it may be related with articular cartilage breakdown.

Objective To investigate the genotype and production of toxin A and B of C .difficile clinical isolates collected from Sydney ,Australia .Methods Sixty‐eight C .difficile clinical isolates were collected from Westmead Hospital ,the University of Sydney ,which were genotyped by using PCR‐ribotyping ,and toxin A ,B coding gene tcdA ,tcdB were detected by using PCR meth‐od .Results Thirty‐one PCR‐ribotypes (RTs) were confirmed in the 68 C .difficile clinical isolates ,RT014 (19 .1% ) and RT002 (11 .8% ) were the common genotypes .Sixty‐four of 68 (94 .1% ) isolates contained tcdA and tcdB for toxin A and B .Conclusion The common prevalent PCR‐ribotypes of C .difficile were RT014 and RT002 in Sydney ,most of the C .difficile clinical isolates contained toxin A and B .

Objective To investigate the genotype and variance of toxin associated genes of moxifloxacin‐resistant Clostridium difficile clinical isolates in Sydney .Methods Twenty‐two moxifloxacin‐resistant Clostridium difficile clinical isolates were collected from Sydney ,which were genotyped by using sequencer capillary gel electrophoresis based PCR‐ribotyping ,and toxin A and B cod‐ing gene tcdA and tcdB ,and binary toxin coding gene cdtA and cdtB were detected by using PCR method .Toxin regulator gene tc‐dC was analyzed by using PCR‐sequencing ,and was aligned with reference sequence of VPI 10463 (Genbank accession number :X92982) ,and the tcdC sequence types of all 22 isolates were identified by using blast tool in NCBI .Results Twenty‐one isolates were genotyped as hypervirulent PCR‐ribotypes 027 (RT027) ,and one isolate as RT078 ;all 22 isolates contained tcdA and tcdB for toxin A and B and cdtA and cdtB for binary toxin (tcdA+ tcdB+ cdtA+ cdtB+ ) .The tcdC sequence types of the 21 RT027 i‐solates belong to sc1 ,and that of the one RT078 isolate belongs to WA39 .Compared with tcdC reference sequence of VPI 10463 ,a consecutive 18 bp deletion (nt341 to 379) and one nucleotide deletion at position 117 were found in the 21 RT027 isolates ,and a consecutive 39 bp deletion (nt330 to 368) and one nucleotide mutation at position 184(C> T) were found in the one RT078 isolate . Conclusion Clostridium difficile hypervirulent RT027 was the common moxifloxacin resistant genotype ;Clostridium difficile hy‐pervirulent RT027 and RT078 clinical isolates contained genes for toxin A and B and binary toxin ,and contained gene sequence mu‐tation in toxin regulator gene tcdC .

BACKGROUND AND PURPOSE: Whether or not migraine can cause cumulative brain alterations due to frequent migraine-related nociceptive input in patients is largely unclear. The aim of this study was to characterize longitudinal changes in brain activity between repeated observations within a short time interval in a group of female migraine patients, using resting-state functional magnetic resonance imaging. METHODS: Nineteen patients and 20 healthy controls (HC) participated in the study. Regional homogeneity (ReHo) and functional interregional connectivity were assessed to determine the focal and global features of brain dysfunction in migraine. The relationship between changes in headache parameters and longitudinal brain alterations were also investigated. RESULTS: All patients reported that their headache activity increased over time. Abnormal ReHo changes in the patient group relative to the HC were found in the putamen, orbitofrontal cortex, secondary somatosensory cortex, brainstem, and thalamus. Moreover, these brain regions exhibited longitudinal ReHo changes at the 6-week follow-up examination. These headache activity changes were accompanied by disproportionately dysfunctional connectivity in the putamen in the migraine patients, as revealed by functional connectivity analysis, suggesting that the putamen plays an important role in integrating diverse information among other migraine-related brain regions. CONCLUSIONS: The results obtained in this study suggest that progressive brain aberrations in migraine progress as a result of increased headache attacks.
Brain Stem ; Brain* ; Female ; Follow-Up Studies ; Headache ; Humans ; Longitudinal Studies* ; Magnetic Resonance Imaging ; Migraine Disorders* ; Putamen ; Somatosensory Cortex ; Thalamus