流行性乙型脑炎病毒NS5蛋白的表达

目的 在大肠杆菌中表达并纯化流行性乙型脑炎病毒NS5蛋白,为进一步研究乙脑病毒NS5蛋白的结构与功能奠定基础。方法 根据乙脑病毒NS5蛋白的基础序列,设计两对PCR引物,采用RT-PCR法分别扩增乙脑病毒JaOH0566株部分及全长NS5基因。将扩增产物分别克隆到表达载体pQE30。筛选重组质粒,转化大肠杆菌表达重组蛋白。重组蛋白经金属柱亲和层析纯化。结果与结论 获得了含部分及全长乙脑病毒NS5基因的重组质粒,重组质粒经限制性酶双酶切、DNA序列分析及PCR扩增筛选证实。在大肠杆菌中表现出了带6个组氨酸头的部分及全长的重组乙脑病毒NS5蛋白。经SDS-PAGE及Western-blot分析证实表达蛋白确定是重组乙脑病毒NS5蛋白。     

DETECTION OF ANTIRIBOSOMAL P PROTEIN ANTIBODIES IN PATIENTS WITH SYSTEMIC SCLEROSIS

This study investigated the prevalence and clinical significanceof anti-ribosomal P protein (anti-P) antibodies in patientswith systemic sclerosis (SSc). Serum samples from 150 patientswith SSc were examined by indirect immunofluorescence, ELISAand immunoblotting. Anti-P antibodies were detected in four(3%) patients with SSc. Three of the four patients showed SSc/SLE(systemic lupus erythematosus) overlap syndrome, but psychiatricdisorders were not observed in these patients. By longitudinalimmunoblotting analysis one patient, who was initially diagnosedwith SSc, later developed anti-P antibodies along with clinicalmanifestations of SLE. Our data suggest that anti-P antibodiesare uncommon in SSc and that the presence of anti-P antibodiesin patients with SSc indicates an overlap with SLE. KEY WORDS: Anti-ribosomal P protein antibodies, Systemic sclerosis, Systemic lupus erythematosus     

ULTRASOUND MEASUREMENT OF SKIN THICKNESS IN SYSTEMIC SCLEROSIS

Sclerotic skin change in systemic sclerosis (SSc) usually accompaniesincreased skin thickness. In order to quantify the cutaneouschanges and to clarify the changes in the ‘uninvolved’skin in systemic sclerosis (SSc), we measured the skin thicknesson the chest, the forearms and the hands of 79 patients withSSc and 81 healthy controls with a B-mode ultrasound (30 MHz)apparatus. The thickness of the ‘uninvolved’, aswell as the ‘involved’ skin in patients with SScwas significantly greater than that of healthy controls. Increasedskin thickness on the forearms and/or the hands showed a 64.6%sensitivity and a 100% specificity for SSc. These results indicatedthat the skin which appears to be ‘uninvolved’ inpatients with SSc is already pathologic, as shown by increasedthickness. Moreover, measurement of skin thickness may be beneficialin the diagnosis of this disease at an early stage. KEY WORDS: Systemic sclerosis, Skin thickness, ‘Uninvolved’ skin, Ultrasound measurement     

New hammock hypothesis-based method for the treatment of stress urinary incontinence: the first 29 urethral supports with a small fascial patch

AIM: We have developed a new method requiring no manipulations of the retropubic passages, and carried out a retrospective study to assess the effectiveness, safety and early results of this new procedure in the treatment of female stress urinary incontinence. METHODS: From January 2001 to September 2003, 29 patients underwent our new surgery for the treatment of SUI. A 3 x 2 cm vertical rectangle was harvested from the rectus fascia. A convex horizontal incision was made over the mid urethra and bladder neck. The retropubic space was entered to the urethropelvic ligament spreading toward the junctions between urethropelvic ligament and tendinous arc. One suture was made at the point lateral to the bladder neck. Another suture was made behind the pubic bone. These two sutures were sutured again and tied with the short side end of the harvested fascia. The same procedure was performed on the other side. RESULTS: Of 29 patients, 26 (89.7%) were cured of stress urinary incontinence and 3 were improved. None of the patients had difficulty in voiding after the operation. Maximum flow rates were almost the same before and after surgery. None of the patients had any adverse events. CONCLUSION: This procedure, which requires no manipulations of the retropubic passages, is safe and effective for the treatment of stress urinary incontinence.     

Tiny nodule in the testicle: Case report of a sertoli cell tumor

Sex cord-stromal tumors of the testis are rare. We report on a small Sertoli cell tumor in the testicle. According to published reports, a nodular lesion on the testicle has a variety of differential diagnoses. Preoperatively, it is very difficult to differentiate between a tumorous lesion and an inflammatory change. When a tiny nodule in the testicle is encountered, we propose limited, testicular-sparing surgery according to the frozen section diagnosis.     

Visualization of palisade veins in esophageal varices by endoscopic color Doppler ultrasonography

Background: Endoscopic color Doppler ultrasonography (ECDUS) is a useful modality for obtaining color flow images of esophageal varices. Levovist is a microbubble echo‐enhancing agent that improves Doppler ultrasound examination. This study is designed to evaluate the usefulness of ECDUS using Levovist in diagnosing palisade veins of esophageal varices. Methods: The study involved 67 patients with esophageal varices using ECDUS. All 67 patients received Levovist intravenously at a concentration of 300 mg/mL. A 7.5‐mL dose of the contrast agent was injected at a slow infusion rate of 1 mL/min. We compared vessel images detected with precontrast with those detected by enhanced ECDUS. Results: Color flow images of palisade veins were obtained in 16 (23.9%) of the 67 patients with precontrast ECDUS. Vessel images of palisade veins were detected in 15 of 61 F2 type varices (24.6%) and in one of six F3 varices (16.7%). The color flows of these vessels showed a continuous wave on fast‐Fourier transform analysis. Sixteen palisade veins had velocities in the 3.3 cm/s?11.6 cm/s range. Color flow images of palisade veins were obtained in 27 (40.3%) of the 67 cases by enhanced ECDUS using Levovist. Palisade veins could be delineated after Levovist contrast in 11 patients who could not be detected on precontrast ECDUS. After Levovist contrast, color flow images detected with precontrast ECDUS were enhanced in all patients. Conclusion: Endoscopic color Doppler ultrasonography with Levovist contrast can improve the diagnostic quality of the palisade veins in esophageal varices.     

The usefulness of ureterorenoscopic examination in evaluation of upper tract disease

AIM: The objective of the present study was to clarify the indications, usefulness and limitations of ureterorenoscopy. MATERIAL AND METHODS: From January 1998 to June 2004, 72 consecutive patients (48 men and 24 women) with a mean age of 66 years (range, 27-83 years) underwent ureterorenoscopy to diagnose upper urinary tract tumors (UUT). Median follow-up was 24 months (range, 1-73 months). Patients were divided into four subgroups by voided urine cytology and preoperative radiographic findings. Group A (n=11, 15.3%), positive voided urine cytology and positive preoperative radiographic findings; group B (n=5, 6.9%), positive cytology and negative radiographic findings; group C (n=48, 66.7%), negative cytology and positive radiographic findings and group D (n=8, 11.1%), frank hematuria originating from the UUT but negative cytology and negative radiographic findings. We compared the findings of ureterorenoscopic examination and biopsy with the results of retrograde pyelography and cytology of upper tract urine. For each examination, the following diagnostic indices were assessed: sensitivity, specificity, positive-predictive-value (PPV) and negative-predictive-value (NPV) and accuracy. Statistical analysis was performed using McNemar's test. RESULTS: For ureterorenoscopy, sensitivity was 94%, specificity 59%, PPV 72%, NPV 92% and accuracy 76%. For biopsy, sensitivity was 77%, specificity 100%, PPV 100%, NPV 80% and accuracy 88%. Accuracy of ureterorenoscopy tended to be superior to that of retrograde pyelography. Ureterorenoscopy was most useful in the group which consisted of 48 patients (66.7%) with negative voided urine cytology and positive preoperative radiographic findings. This group was the only group in which accuracy of ureterorenoscopic biopsy was superior to that of urine cytology, significantly (P=0.03). CONCLUSION: Results indicated that ureterorenoscopy is most suitable and gives superior accuracy in patients with positive radiographic findings and negative voiding cytology. Ureterorenoscopic biopsy of the upper urinary tract would provide useful information when considering therapeutic strategies, such as nephron-sparing management.