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.     

Can rhBMP-2 Containing Collagen Sponges Enhance Bone Repair in Ovariectomized Rats?: A Preliminary Study

With an aging population the frequency of postmenopausal fractures is increasing. Methods to enhance the repair of osteoporotic bone repair therefore become more important to reduce the society burden of care. We asked if absorbable collagen sponges containing recombinant human bone morphogenetic protein-2 (rhBMP-2) have the potential to enhance bone repair. We randomly assigned 40 rats into the ovariectomy and sham operation groups. A segmental defect was created in the right tibia 12 weeks after ovariectomy. rhBMP-2-containing absorbable collagen sponges were implanted into the defect in half of the animals in each group. We analyzed radiographs and histological sections and performed three-point bending tests to assess repair. Radiological scores in the rhBMP-2 applied rats were higher than those in controls at the end of 8 weeks after tibial osteotomy. The specimens failed under higher loads in the rhBMP-2-applied groups and histology revealed a higher fracture healing score, including callus formation, bone union, marrow changes, and cortex remodeling. We observed no adverse tissue responses such as fibrous connective tissue formation and inflammatory cellular infiltration. rhBMP-2 in absorbable collagen sponges enhanced bone repair in segmental tibial defects of ovariectomized rats. The sponges with rhBMP-2 appeared to enhance bone repair.     

Free nerve endings and morphological features of the ligamentum capitis femoris in developmental dysplasia of the hip

A conflict exists on whether the ligamentum capitis femoris has the neuro-morphological structures required for nociception or proprioception of the hip joint. Therefore, we investigated the morphological features and the presence of mechanoreceptors in 24 ligamentum capitis femoris biopsies obtained at open reduction in patients with developmental dysplasia of the hip. Of these 24 hips, 16 were completely dislocated and eight were subluxated. The mean age was 33.8 months (range 13-52 months) at the time of surgery. En bloc ligamentum capitis femoris and pulvinar were taken for biopsy specimen. Ligamentum capitis femoris was dissected and the weight of each ligament was determined using a highly sensitive balance. Specimens were stained with hematoxylin and eosin and Masson trichrome for routine histolopathological evaluation and examined immunohistochemically using monoclonal antibody against S-100 protein. All specimens were graded on a four-grade system according to the amount of coarse-thick collagen bundles and hyalinization. The mean number and type of mechanoreceptors of each specimen were recorded. When the mean age, the patient's weight and the ligamentum capitis femoris weight of each group (completely dislocated vs. subluxated) were compared, there were no significant differences. In the ligamentum capitis femoris of the dislocated hips, the cells were irregularly distributed, had different shapes, and appeared to be in different stages of functional activity. The collagen fiber bundles were thicker than in the subluxated hips, distributed and of varied thickness. The elastic fibers of the dislocated hips were thicker and more numerous than those in the subluxated hips. We found a significant difference between the two groups with regard to the grade of collagen and hyalinization of ligamentum capitis femoris (P<0.004). We found type IVa, free nerve endings in 16 of 24 samples of ligamentum capitis femoris. The 66.6% presence of free nerve endings in the ligamentum capitis femoris suggests a role in nociception/proprioception of the hip in developmental dysplasia of the hip. Interestingly, the percentage and the mean numbers of free nerve endings containing ligamentum capitis femoris were similar in completely dislocated hip group and the subluxated group (62.5 vs. 75%, 12.13+/-9.07 vs. 9.37+/-9.24, respectively). We conclude that the morphological features of ligamentum capitis femoris are influenced by the severity of developmental dysplasia of the hip, whereas the distribution of free nerve endings are not influenced.     

Mini-open versus closed reduction in titanium elastic nailing of paediatric femoral shaft fractures: a comparative study

The purpose of this study is to compare retrospectively intraoperative fluoroscopy time and clinical-radiological results in pediatric femoral shaft fractures treated with titanium elastic nailing (TEN), with a mini-open "blind-hand" technique versus closed reduction. The study included 87 children (18 girls and 69 boys) who underwent surgical treatment with TEN for femoral shaft fractures. Patients were divided into two groups. Group 1 included 42 patients (mean age : 83 +/- 2.7 years) treated with a mini-open "blind-ha nd" technique (a 2-3 cm lateral incision at the level of the fracture ; reduction achieved with one or two fingers, without visualization of the fracture). Group 2 consisted of 45 patients (mean age: 8.8 +/- 2.6 years) treated with a closed reduction technique. Duration of surgery and intraoperative fluoroscopy time were recorded in both groups. Clinical and radiologic results were assessed using the TEN scoring system after mean follow-up periods of 213 +/- 5.8 months and 193 +/- 5.6 months in group 1 and group 2, respectively. Mean duration of surgery was 31.7 +/- 7.6 and 52.1 +/- 14.4 minutes, and mean fluoroscopy time 32.9 +/- 22.1 and 75.1 +/- 31.5 seconds in group 1 and group 2, respectively. Both surgical and fluoroscopy time were significantly longer in group 2 (p < 0.001). There was no significant difference between the two groups in terms of clinical and radiological results. All fractures healed with solid union, and there was no complication that was expected to cause permanent disability. Although successful clinical and radiological results were obtained with both techniques, duration of surgery and intraoperative fluoroscopy time were significantly higher in the closed reduction group 2. We suggest the "blind-hand" technique as an alternative to closed reduction to prevent extensive intraoperative radiation exposure and to decrease the length of the surgical procedure.     

Quantity not composition of dietary fats represents the dominant contributor to experimental obesity: Relevance to human pathophysiology

Plant fats are low in saturated fats but high in unsaturated fats compared to animal fats, and are supposedly less obesogenic. This study compared the obesogenic effects of plant and animal derived fatty diets in Wistar rats. Rats of each gender were divided into three dietary (standard chow (SC), high fat diet rich in animal fat (HFDaf) and a high fat diet rich in plant fat (HFDpf)) groups of ten each and fed for 17 weeks. Anthropometric, Adiposity and nutritive variables were assessed using standard methods. Comparing HFDpf to HFDaf: Abdominal circumference (AC),initial feed intaken (IFI), final feed intake(FFI), final body weight (FBW), white adipose tissue (WAT) were increased but brown adipose tissue (BAT) decreased in male rats fed with HFDpf; also, there were increased body length, IFI, FFI but decreased AC, FBW, BAT in female rats fed with HFDpf. Comparing male to female rats: Thoracic circumference, IFI, FFI, energy intake were increased while Adiposity index decreased across diet groups in male rats; the AC, FBW increased while WAT, BAT decreased in HFDpf fed group, also, BAT was increased but AC, FBW decreased in HFDaf fed group in male rats. Palatability and high feed efficiency of consumed diets were more associated with obesogenic risk than just the level of saturation. Therefore, Obesogenic effects of fatty diets in both genders is more dependent on the quantity (amount) of fatty diet consumed than the dietary fat composition alone.     

Platelet-leukocyte interaction: selective binding of thrombin- stimulated platelets to human monocytes, polymorphonuclear leukocytes, and related cell lines

The association of platelets with leukocytes was investigated, using gel-filtered platelets stimulated with thrombin and then fixed with formaldehyde. Evidence is presented that stimulation of gel-filtered platelets with low concentrations of thrombin (0.01 to 0.1 U/mL) induces the expression of surface determinants interacting strongly with monocytes and polymorphonuclear leukocytes (PMNs) but only weakly with lymphocytes. Both monocyte-platelet binding and PMN-platelet binding occurred at 37 degrees C and more intensively at 0 degrees C; it was Ca2+-dependent and was unaffected by the addition of sodium azide. The binding also occurred with the monocytoid cell lines HL 60 and U 937 in exponential growth and was much less two days after induction of terminal differentiation by phorbol myristate acetate (PMA). No other tested cell lines (B cells, T cells, and myeloid cells) bound thrombin-stimulated platelets. Monocyte-derived macrophages kept in culture for one week also exhibited reduced binding of thrombin- stimulated platelets. IgG and fibronectin could be ruled out as ligands that mediate binding.     

Tetra-oligodactyly with bilateral aplasia and hypoplasia of long bones of upper and lower limbs: A variable manifestation of the syndrome of ectrodactyly with tibial aplasia

We resent, a family manifesting a variation of the syndrome of ectrodactyly with tibial aplasia. The principal case in the family showed the most severe bilateral skeletal malformations of this syndrome. The hand changes of this case (tetra-oligodactyly with missing 5th rays) and of a relative (oligodactyly with the last 3 rays being affected) reflected a variable manifestation of ectrodactyly. Additionally, a review of the relevant literature is presented for further delineation of various aspects of this syndrome.     

循证护理应用才精神科患者的效果评价

目的运用循证医学的思维方法，探讨在精神科护理工作中开展循证护理所达到的临床康复效果。方法将120例男性住院精神分裂症患者随机分为研究组和对照组各60例，研究组接受为期2个月的以循证理论为依据的综合性心理护理和技能训练，对照组常规给予一般支持性心理护理。采用护士用住院患者观察量表（NOSIE）、简明精神病评定量表（BPRS）、社会功能缺陷筛选量表（SDSS）、精神科症状自评量表（SCL-90）和自制的患者满意度／舒适度调查表作为评价工具，比较2组干预后的效果。结果入组时，所有量表评分2组间比较差异均无统计学意义（P〉0.05），经过2个月的治疗和循证干预后，2组间各量表评分比较，研究组优于对照组，差异均有统计学意义（P〈0.05）。结论循证护理可促进科学、有效的精神科护理实践活动，有助于促进精神分裂症患者的临床治疗和康复，为科学有效的临床护理决策提供依据和工作方法。