真核表达人呼吸道合胞病毒F蛋白的纯化方法

目的 真核表达人呼吸道合胞病毒(human respiratory syncytial virus,SV)融合蛋白(fusion protein,),并完成蛋白纯化及纯度测定.方法 根据编码F蛋白的基因序列设计引物,CR方法扩增出3'端带His标签的F基因序列,克隆入pGEM-T-easy载体,经核酸序列分析后,进一步克隆到pcDNA3.1( )真核表达载体,限制性内切酶鉴定,用脂质体Lipofectamine2000转染COS-7细胞,2 h后再用Westem blot检测目的蛋白的表达.Ni柱亲和层析纯化COS-7细胞表达的F蛋白,高效毛细管电泳分析纯化后蛋白纯度.结果 核酸序列分析证实获得带His标签的RSV F基因序列,没有发生无义突变.转染COS-7细胞后,利用Western blot方法检测到F蛋白的特异性条带,纯度达99%以上.结论 初步建立了真核表达RSV F蛋白的纯化方法,为进一步优化RSV F蛋白制备条件及单克隆抗体及诊断试剂等研究奠定了基础.     

Nosocomial and community acquired uropathogenic isolates of Proteus mirabilis and antimicrobial susceptibility profiles at a university hospital in Sub–Saharan Africa

ObjectiveTo ascertain antimicrobial susceptibility profile of Proteus mirabilis (P. mirabilis) from clinical urine specimens at a university hospital in the spate of its recorded increasing resistance patterns.MethodsThe study was retrospective in nature. Data generated from urine cultures of patients at University of Calabar Teaching Hospital for a period of five years (2004–2009) were compiled. Relevant information obtained were age and gender of patients, organisms recovered and their antibiotic susceptibility patterns. P. mirabilis was identified using standard laboratory procedures.ResultsP. mirabilis showed the highest resistance against ampicillin, cloxacillin, amoxicillin, tetracycline, co-trimoxazole, erythromycin and chloramphenicol (100%–37.2%) while colistin, ofloxacin, ciprofloxacin, ceftriaxone, nalidixic acid and nitrofurantoin recorded the highest activity (59.1%–96.9%) with no drug recording 100% activity. The resistance of the nosocomial isolates of the organism were significantly higher than the community acquired isolates against that of the common antibiotics in use (P<0.05).ConclusionsExtreme caution should be exercised in antibiotic administration in hospital setting and the potential benefits adequately assessed while control of nosocomial infections be given a priority so as to limit the spread of resistant bacteria.     

Progression of Metabolic Syndrome in Renal Transplant Recipients

Background

Metabolic syndrome, which is closely related to insulin resistance, is highly prevalent in renal transplant recipients.

Purpose

We aimed to investigate prevalence, risk factors, and progression of metabolic syndrome in renal transplant recipients.

Methods

One hundred fifty-eight renal transplant recipients who had been on transplantation for more than 1 year and 79 age-sex matched healthy controls were included in the cross-sectional phase of the study. We measured baseline characteristics, blood pressure, fasting blood glucose, and lipid profiles and we defined metabolic syndrome using the National Cholesterol Education Program Adult Treatment Panel III criteria. One hundred twenty-four renal transplant recipients were eligible for the second evaluation after 22.9 ± 3.8 months. Metabolic syndrome prevalence and homeostasis model assessment insulin resistance levels were evaluated during the follow-up period.

Results

Overall, metabolic syndrome was present in 34.2% of the patients and 12.7% of the controls at the cross-sectional phase of the study (P = .000). Only the hypertension component of metabolic syndrome was significantly increased in patients compared to controls (P = .000). Pretransplantation weight and body mass index were significantly higher in patients who had metabolic syndrome (P = .000). During the follow-up period, prevalence of metabolic syndrome did not change (P = .510); however, body mass index and blood pressure increased and the high density lipoprotein cholesterol component of metabolic syndrome decreased (P = .001). We did not find any significant difference in glomerular filtration rate change among patients with and without metabolic syndrome (−2.2 ± 11.36 vs −6.14 ± 13.19; P = .091). Glucose metabolism parameters including hemoglobin A1c, insulin, and homeostasis model assessment insulin resistance were disturbed in patients with metabolic syndrome (P = .000, P = .001, P = .002, respectively).

Conclusion

Metabolic syndrome is highly prevalent in renal transplant recipients and closely associated with insulin resistance. The prominent criterion of metabolic syndrome in patients seems to be hypertension, especially high systolic blood pressure. The identification of metabolic syndrome as a risk factor may yield new treatment modalities to prevent it.     

Ultrasonographic Maturation of Native Arteriovenous Fistulae: A Follow-Up Study

Background/aims. Ideal time needed for arteriovenous fistula (AVF) maturation is still controversial. In this study, we aimed to investigate the natural course of AVF maturation and also investigated the factors affecting AVF maturation. Methods. We studied 31 (21M/10F, mean age 55.8 ± 16.2) chronic renal failure patients. We evaluated the patients with color Doppler ultrasound examination before the fistula operation, at the first day, and at the first, second, third, and sixth months. Radial artery (RA) diameter, flow velocity, flow, resistance index, fistula vein diameter, flow velocity, and flow were measured. Results. Patency rates at the first post-operative day and the sixth month were 87.1% and 67.1%, respectively. Cephalic vein flow was 451.2 ± 248.6 mL/min at the first month and 528.6 ± 316.5 mL/min at the sixth month. Baseline RA diameter was lower in failing fistulas than that of patent fistulas. Failing fistulas were more common in women. Conclusion. Blood flow was enough for hemodialysis at the end of the first month. However, fistula maturation had continued until the end of the study; women and patients with low RA diameter are particularly prone to fistula failure. Therefore, especially in these patients, AVF must be created at least three or four months before the predicted hemodialysis initiation time.     

Antioxidant effects of vitamin D on lacrimal glands against high dose radioiodine-associated damage in an animal model

Purpose: To evaluate antioxidant effects of active vitamin D (calcitriol) against high-dose radioiodine (RAI) therapy-associated damage of lacrimal gland.

Materials and methods: Wistar albino rats were used and divided into three groups randomly (n?=?12/group). The first group was appointed as the negative control group and received no RAI or medication. The second group was appointed as the positive control group that only received 3?mCi/kg (111 MBq/kg) RAI via gastric gavage and the last group was the treatment group that received 3?mCi/kg RAI via same method and calcitriol (200?ng/kg/day) via intraperitoneal administration. Seven days after RAI administration, bilateral intraorbital (IG), extraorbital (EG) and Harderian (HG) glands were removed for the evaluations of histopathologic, tissue cytokine, total oxidant status (TOS) and total antioxidant status (TAS).

Results: RAI led to significant increase in tissue TOS, TNF-α, IL-6 levels and significant decrease in IL-10 and TAS levels (p?p?p?p?=?0.049), periductal fibrosis in EG and HG (p?=?0.049 and 0.038, respectively), abnormal cell outlines in EG and HG (p?=?0.020 and 0.011, respectively) and variation in cell size in the IG and the HG (p?=?0.003 and 0.049 respectively).

Conclusions: RAI caused significant oxidative stress and inflammation in lacrimal glands. Vitamin D demonstrated potent anti-inflammatory, antioxidant and radio-protective effects on lacrimal glands in histopathologic, tissue cytokine and oxidant/antioxidant level evaluations.     

Relative erythrocytosis of patients with end stage renal failure undergoing CAPD

BACKGROUND AND AIM: Although the majority of patients with end stage renal failure have anemia, some have relative erythrocytosis. Patients treated with continuous ambulatory peritoneal dialysis (CAPD) having relative erythrocytosis were studied in order to determine the factors that would be responsible. METHODS: Nine out of 89 CAPD patients (10%) were identified as having relative erythrocytosis. Age-, sex- and duration of disease-matched eight patients undergoing CAPD were taken as control. Beside factors of etiologies of renal failure, smoking, renal cysts, viral hepatitides, residual renal function, the adequacy of CAPD, nutritional status, hypertension, serum levels of erythropoietin, IL-1, IL-6, TNF-, and IGF-1 levels were also investigated. RESULTS: Relative erythrocytosis occurred most often in diabetic and amyloidosis patients. None of the parameters studied were found to be significantly different between groups. During 2-year follow-up, although statistically non-significant, patients having relative erythrocytosis seemed to have higher mortality rate due to vascular complications. CONCLUSION: No single factor seemed to explain erythrocytosis in patients undergoing CAPD. Being diabetic or with amyloidosis may increase the risk.     

Nephrotic syndrome associated with agnogenic myeloid metaplasia

Extramedullary hematopoiesis being an important feature of agnogenic myeloid metaplasia (AMM), a chronic myeloproliferative disease of clonal origin, may affect the kidneys, but this condition is usually asymptomatic. Until now, there is only one reported case of nephrotic syndrome associated with AMM. We present a patient with AMM who had nephrotic syndrome and whose renal biopsy revealed membranous glomerulonephritis together with renal extramedullary hematopoiesis.