流行性乙型脑炎病毒结构蛋白编码基因的重组构建

目的:建立流行性乙型脑炎病毒(JEV)C、prME,E等结构蛋白编码基因克隆.方法:病毒感染C6/36细胞,RT-PCR法依次获取该病毒结构蛋白基因并进行DNA测序分析,PCR克隆法将PCR产物分别构建于真核表达载体pcDNA 3.1的BamHI与EcoRI酶切位点并依次命名为pJC、pJME与pJE,通过DNA测序、电泳以及限制性内切酶分析加以鉴定.结果:JEV C、prME与E基因片段分别为380、2001以及1 500bp,各基因测序结果与发表的JEV JaGAr-01株相对应序列相一致,从重组质粒释出的插入子分别与各基因大小相符合.结论:pJC、pJME与pJE重组质粒分别含有C、prME与E蛋白编码基因.     

Molecular cloning and complete nucleotide sequence of the genome of Japanese encephalitis virus Beijing-1 strain

The genomic RNA of the Japanese encephalitis virus (JEV) Beijing-1 strain was reversely transcribed and the synthesized cDNA was molecularly cloned. Six continuous cDNA clones that cover the entire virus genome were established and sequenced to determine the complete nucleotide sequence of the JEV RNA. The precise genomic size was estimated as 10,965 bases long. With flanking 95 bases at the 5 and 583 bases at the 3 non-coding regions, one long open reading frame (ORF) was revealed encoding a virus polyprotein with 3,429 amino acid residues. Because of sequence homologies observed between JEV and other flaviviruses, the genome organization of JEV appears to be identical with other flaviviruses. Genetic variation detected among flavivirus genomes is consistent with the established serological relatedness between JEV and other members of flaviviruses. The secondary structure of the JEV genome is deduced and discussed concerning its involvement in genome replication.     

Antihypertensive constituents in Sanoshashinto

It has been reported that Sanoshashinto (SanHuangXieXinTang, 三黃瀉心湯), which is composed of Rhei Rhizoma, Scutellariae Radix, and Coptidis Rhizoma, exhibits vasorelaxant effects in vitro and lowers blood pressure of patients. Based on this discovery, in this study, a mixture containing those three materials and combinations of them were extracted with methanol, and the extracts were fractionated into different parts. Effects of all extracts and fractions on high concentration of potassium chloride (High K+)- or noradrenaline (NA)-induced contractions of isolated rat aortic rings or helical strips were examined. Qualitative and quantitative HPLC analyses of the extracts and the fractions revealed that the contents of baicalin and berberine in Sanoshashinto methanol extract (SHXXTM) were higher than those of the other constituents. All pharmacological and HPLC data were analyzed by principal component analysis (PCA) software and the results indicated that baicalin, berberine, palmatine, baicalein, and wogonoside contributed significantly to the pharmacological activity. Furthermore, spontaneously hypertensive rats (SHRs) that were orally given SHXXTM or a baicalin–berberine combination showed significantly reduced increase in the rate of systolic blood pressure (SBP) compared to the control group. These findings suggested that Sanoshashinto has significant vasorelaxant effects in vitro and antihypertensive effects in vivo, and baicalin and berberine, which were the principal constituents of Scutellariae Radix and Coptidis Rhizoma, were the main antihypertensive constituents in Sanoshashinto. It was speculated that baicalin and berberine produced vasorelaxant effects by activating the NO/cGMP pathway and that the BKCa channel and the DAG/PKC/CPI-17 pathway were also involved.     

Clonality analysis of lymphoproliferative disorders in patients with Sjögren's syndrome

The aim of this study was to clarify the nature of the clonal lymphocyte infiltration in Sjögren''s syndrome (SS) patients associated with lymphoproliferative disorders. We examined B cell clonality in lymphoproliferative tissues from six primary SS patients associated with lymphoproliferative disorders or lymphoma by cloning and sequencing of the gene rearrangement of the immunoglobulin heavy chain complementarity determining region 3 (IgVH–CDR3). Three patients with sequential observation showed progressional clonal expansion with the presence of the same subclone in different tissues during the course of disease. Among them, one patient developed mucosa-associated lymphoid tissue (MALT) lymphoma in glandular parotid. The other three SS patients concomitant with malignant B cells lymphomas showed different clonal expansion of B cells between nodal sites and salivary glands. The cloanality analysis indicated that monoclonal B cell population could spread from one glandular site to another site during the course of SS, suggesting that the malignant clone may arise from the general abnormal microenvironment, not restricted to the glandular tissue, in some SS patients.     

Does unilateral total hip arthroplasty improve pain and function in the non-operative hip joint?

European Journal of Orthopaedic Surgery & Traumatology - Some reports suggested that the status of the opposite-side hip affects clinical outcomes of unilateral total hip arthroplasty (THA) for...     

Laterality of radiographic osteoarthritis of the knee

There are few reports of the laterality in radiological knee osteoarthritis (ROA). This study aimed to evaluate laterality in terms of the minimum joint space width (mJSW) and osteophyte areas (OFs) in a cross-sectorial general population screen and elucidate the association between laterality and risk of osteoarthritis. We enrolled 330 participants (mean age 64.6 years) and examined the presence of ROA (Kellgren–Lawrence grade?≧?2) laterality in terms of the mJSW and OF on the medial tibia using auto-measuring software. Moreover, we examined the association between laterality and leg dominance. The right and left medial mJSWs were 4.02?±?0.98?mm and 4.05?±?1.01?mm, respectively, showing no laterality; the laterals were also similar. The participants who had osteophytes ≥1?mm² in the right, left, and bilateral knees were 15, 37, and 57 respectively, with osteophytes being significantly more common in the left knee. The OF was significantly larger in the left knee. Conversely, the medial and lateral mJSWs and OF did not differ according to leg dominance. The prevalence of ROA was higher and the OF was more pronounced in the left knee. However, the mJSW showed no laterality. Additionally, the mJSW and OF showed no differences according to leg dominance.     

19F NMR spectrometric determination of the partition coefficients of some fluorinated psychotropic drugs between phosphatidylcholine bilayer vesicles and water

A simple 19F NMR spectrometric method was proposed for the determination of the partition coefficients of fluorinated psychotropic drugs, trifluoperazine (TFPZ), flunitrazepam (FNZ) and flurazepam (FZ) between phosphatidylcholine (PC) bilayer of small unilamellar vesicles (SUVs) and water (buffer). Each 19F NMR spectrum of these drugs in the presence of PC SUV showed a single signal accompanying a PC concentration-depending shift change and broadening, which indicated a fast exchange of these drugs between the water phase and the PC bilayer of SUV. From the relationship between the 19F chemical shift change (Deltadelta) of each drug and the PC concentration, the molar partition coefficients (K(p)'s) were calculated and obtained with a good precision of RSD below 6%. The fractions of the partitioned drugs calculated by using the obtained K(p)-values were in a good agreement with the experimental values. The results demonstrate that the 19F NMR method can be usefully applied to the determination of partition coefficients of many drugs having fluorine atom(s) without any separation procedure, especially for drugs which do not have absorption in the ultraviolet or visible region, or those having absorption but show insignificant spectral changes according to their incorporation to PC bilayers (e.g. FNZ).     

Change of barometric pressure influences low back pain in patients with vacuum phenomenon within lumbar intervertebral disc

The present study investigates whether changes in barometric pressure influence low back pain in patients with vacuum phenomenon within lumbar intervertebral discs. Twenty-four patients with low back pain were examined: 12 with vacuum phenomenon within the intervertebral discs (VP [+] group) and 12 without the vacuum phenomenon (VP [-] group). All patients consented to an evaluation of low back pain in a hyperbaric oxygen chamber. In the VP (+) group, low back pain became more severe in one patient when atmospheric pressure was elevated, and in eight patients when atmospheric pressure was decreased. In the VP (-) group, two patients experienced more severe low back pain when atmospheric pressure was decreased. Our study demonstrated that low back pain might be aggravated by atmosphere depression in patients with lumbar disease associated with the vacuum phenomenon.     

Outcome of late surgical treatment in patients with incomplete paraplegia due to spinal degenerative diseases

STUDY DESIGN: Retrospective analysis. OBJECTIVE: To assess the outcome of late surgical intervention in patients with incomplete paraplegia due to spinal degenerative diseases. SETTING: Three men and four women with cervical or thoracic spinal degenerative diseases, who preoperatively were unable to walk for more than 6 months in Mie prefecture, Japan. METHODS: Review of clinical records and questionnaire survey regarding the walking ability of patients 2 years after surgery. RESULTS: All seven patients were unable to walk postoperatively. CONCLUSION: A late surgical intervention may not lead to functional recovery in patients with spinal degenerative disease who were unable to walk for at least 6 months.