带血管自体腓骨移植修复胫骨良性骨肿瘤致骨缺损

目的:观察带血管自体腓骨移植修复胫骨中上段良性骨肿瘤致骨缺损的临床应用结果。方法:选择2001-10/2006-12在南京医科大学附属南京第一医院骨科应用带血管腓骨移植修复因良性肿瘤所致胫骨长段骨缺损患者12例,患者均知情同意。肿瘤清除后,采用健侧带血管腓骨皮瓣移植重建患侧胫骨,切取健侧腓骨长度为8￣23cm,平均长度13cm。术后定期随访,根据Enneking评分结果、移植腓骨转归的影像学评价结果和腓骨供区的随访结果进行效果评定,Enneking系统评分根据患者肢体疼痛、功能活动、自我感受、旋转活动度、精细操作、工作能力等6个方面对患肢进行评分,满分30分,26￣30分评为优,21￣25分评为良,16￣20分评为中,11￣15分评为差。结果:全部12例患者均获随访,随访时间6个月￣3年,其中6￣12个月3例,17￣26个月7例,27￣36个月2例,平均19个月。Enneking评分26￣30分7例,20￣25分4例,15￣20分1例。11例关节功能良好,可完成正常生活功能,仅1例因靠近膝关节至关节功能受限,经康复治疗后改善。X射线片均呈现理想的替代形态,无胫骨下端的成角畸形,腓骨均有不同程度的增粗,仅1例胫骨内侧平台塌陷约3.5mm,另11例无明显塌陷。12例患者无供区踝关节外翻畸形及腓总神经支配区感觉异常,腓骨长、短肌肌力四五级,踝关节活动不受限。结论:带血管腓骨移植用于修复胫骨良性骨肿瘤致骨缺损是可行的,但仍需要积累更多的病例数及更长期的随访以观察远期效果。     

Accurate quantitation of the low number of white cells in white cell- depleted blood components

A simple method of accurate quantification of low concentrations of white cells (WBCs) in WBC-depleted blood components was developed by using propidium iodide (PI) to stain the nuclei of WBCs. The method was validated by correlating the PI-determined WBC concentrations with those determined with Coulter S-plus IV counter in units of packed red cells (PRBCs) or random-donor platelets (RDPs). The correlations were linear and had coefficients of 0.99. The sensitivity of PI staining permitted the detection of concentrations of WBCs as low as 1 cell per microL of RDPs or 11 cells per microL of PRBCs. Therefore, PI staining will be useful in investigating the role of transfused WBC-depleted blood components in the prevention of primary alloimmunization to HLA antigens, as well in evaluating various new procedures with high efficiency in depleting WBCs from blood components.     

Renal Disease Associated with Circulating Antineutrophil Cytoplasm Activity

SUMMARY We report our detailed observations on a group of 30 consecutivepatients with renal disease, histologically demonstrated glomerulitisor necrotizing vasculitis, and circulating antineutrophil cytoplasmactivity (ANCA). The annual incidence of ANCA-related renaldisease was seven cases per million population. The sensitivityof serum ANCA for histologically proved glomerular vasculitiswas 79 per cent, with a specificity of 87 per cent. Most patientsresponded to treatment with cyclophosphamide and steroids butcomplications of therapy occurred in just over half the patientsand were serious in 20 per cent. Actuarial survival at 1 yearwas 60 per cent. Age and dialysis requirement did not influenceoutcome and the only identified adverse prognostic factor washypoxic lung disease. We conclude that the association of ANCAwith renal disease is not rare and that positive serology accuratelyidentifies a homogeneous group of patients with similar clinical,histological, and prognostic features. Separation of these patientsinto those with the disease entities of Wegener's granulomatosisand microscopic polyateritis is not straightforward on clinicaland histological criteria, and such a distinction does not yielduseful therapeutic or prognostic information. Simple urinalysisshould always be carried out in patients with undiagnosed systemicillness in order to identify renal disease. ANCA–relatedrenal disease can be treated successfully with cyclophosphamideand steroids and elderly patients should not be excluded fromtreatment, including dialysis if necessary. The ANCA test issimple and quick to perform and, in the appropriate clinicalsetting, accurately identifies patients who may benefit fromimmunosuppressive treatment before a histological diagnosiscan be established.     

Studies of the cell surface of mouse dendritic cells and other leukocytes

The surface of dendritic cells (DC) has been analyzed by means of monoclonal antibodies (Ab) and lactoperoxidase (LPO)-mediated radioiodination. Antigens and other exteriorily disposed polypeptides of purified spleen DC were compared with those of tissue macrophages (M), monocytes, and other bone marrow-derived elements. Quantitative binding studies and autoradiography with (125)I-Ab established that DC expressed high levels of I-A and H-2D, 2 × 10(5) and 1 × 10(5) Ab binding sites per cell, respectively. DC from conventional, germ-free, and specific pathogen-free mice were all rich in Ia. Expression of Ia on B cells was 5-10 percent of that on DC and increased fivefold during lipopolysaccharide mitogenesis. More than 70-90 percent of purified M and monocytes from specific pathogen-free mice were Ia negative, but increased levels of Ia were noted on cells from mice reared under conventional conditions. Thus large amounts of Ia on DC is a constitutive trait, whereas the expression of Ia by other cell types may be governed by the environmental and immunological status of the host. The 2.4G2 Fc receptor Ag was not detected on DC. Peritoneal and spleen M had 10(5) 2.4G2 binding sites/cell, whereas monocytes and lymphocytes were less reactive (1 × 10(4)-3 × 10(4) binding sites/cell). Four other M-related antigens were evaluated. Each had a distinctive tissue distribution and none bound exclusively to M and monocytes. Neither 1.21J (Mac-1) nor F4/80 reacted with DC. Immunoprecipitation studies of externally ((125)I) and biosynthetically ([(35)S]methionine)dabeled cells confirmed the binding data. Sensitive binding assays with (125)I-Ab confirmed previous observations that DC lack Ig and Thy-1. Lyt-1 was also not found on DC, but 5-12 percent of the cells in purified DC preparations expressed both Lyt-2 and Ia. All DC expressed the leukocyte common antigens at levels similar to other leukocytes. The spectrum of surface polypeptides labeled by LPO-mediated iodination was different on M, DC, and lymphocytes. Polypeptides migrating at molecular weights of 155,000, 85,000, and 62,000 appeared to be restricted to DC. These observations establish that the cell surface of DC differs considerably from other leukocytes, including the blood monocyte, and suggest that the DC is part of a unique Ia-rich leukocyte differentiation pathway.     

Antimicrobial susceptibility testing of Gram-positive and -negative bacterial isolates directly from spiked blood culture media with Raman spectroscopy

Patients suffering from bacterial bloodstream infections have an increased risk of developing systematic inflammatory response syndrome (SIRS), which can result in rapid deterioration of the patients’ health. Diagnostic methods for bacterial identification and antimicrobial susceptibility tests are time-consuming. The aim of this study was to investigate whether Raman spectroscopy would be able to rapidly provide an antimicrobial susceptibility profile from bacteria isolated directly from positive blood cultures. First, bacterial strains (n?=?133) were inoculated in tryptic soy broth and incubated in the presence or absence of antibiotics for 5 h. Antimicrobial susceptibility profiles were analyzed by Raman spectroscopy. Subsequently, a selection of strains was isolated from blood cultures and analyzed similarly. VITEK®2 technology and broth dilution were used as the reference methods. Raman spectra from 67 antibiotic-susceptible strains showed discriminatory spectra in the absence or at low concentrations of antibiotics as compared to high antibiotic concentrations. For 66 antibiotic-resistant strains, no antimicrobial effect was observed on the bacterial Raman spectra. Full concordance with VITEK®2 data and broth dilution was obtained for the antibiotic-susceptible strains, 68 % and 98 %, respectively, for the resistant strains. Discriminative antimicrobial susceptibility testing (AST) profiles were obtained for all bacterial strains isolated from blood cultures, resulting in full concordance with the VITEK®2 data. It can be concluded that Raman spectroscopy is able to detect the antimicrobial susceptibility of bacterial species isolated from a positive blood culture bottle within 5 h. Although Raman spectroscopy is cheap and rapid, further optimization is required, to fulfill a great promise for future AST profiling technology development.     

Progressive hearing loss and vestibular dysfunction caused by a homozygous nonsense mutation in CLIC5

In a consanguineous Turkish family diagnosed with autosomal recessive nonsyndromic hearing impairment (arNSHI), a homozygous region of 47.4 Mb was shared by the two affected siblings on chromosome 6p21.1-q15. This region contains 247 genes including the known deafness gene MYO6. No pathogenic variants were found in MYO6, neither with sequence analysis of the coding region and splice sites nor with mRNA analysis. Subsequent candidate gene evaluation revealed CLIC5 as an excellent candidate gene. The orthologous mouse gene is mutated in the jitterbug mutant that exhibits progressive hearing impairment and vestibular dysfunction. Mutation analysis of CLIC5 revealed a homozygous nonsense mutation c.96T>A (p.(Cys32Ter)) that segregated with the hearing loss. Further analysis of CLIC5 in 213 arNSHI patients from mostly Dutch and Spanish origin did not reveal any additional pathogenic variants. CLIC5 mutations are thus not a common cause of arNSHI in these populations. The hearing loss in the present family had an onset in early childhood and progressed from mild to severe or even profound before the second decade. Impaired hearing is accompanied by vestibular areflexia and in one of the patients with mild renal dysfunction. Although we demonstrate that CLIC5 is expressed in many other human tissues, no additional symptoms were observed in these patients. In conclusion, our results show that CLIC5 is a novel arNSHI gene involved in progressive hearing impairment, vestibular and possibly mild renal dysfunction in a family of Turkish origin.     

International variation in GP treatment strategies for subclinical hypothyroidism in older adults: a case-based survey

Background

There is limited evidence about the impact of treatment for subclinical hypothyroidism, especially among older people.

Aim

To investigate the variation in GP treatment strategies for older patients with subclinical hypothyroidism depending on country and patient characteristics.

Design and setting

Case-based survey of GPs in the Netherlands, Germany, England, Ireland, Switzerland, and New Zealand.

Method

The treatment strategy of GPs (treatment yes/no, starting-dose thyroxine) was assessed for eight cases presenting a woman with subclinical hypothyroidism. The cases differed in the patient characteristics of age (70 versus 85 years), vitality status (vital versus vulnerable), and thyroid-stimulating hormone (TSH) concentration (6 versus 15 mU/L).

Results

A total of 526 GPs participated (the Netherlands n = 129, Germany n = 61, England n = 22, Ireland n = 21, Switzerland n = 262, New Zealand n = 31; overall response 19%). Across countries, differences in treatment strategy were observed. GPs from the Netherlands (mean treatment percentage 34%), England (40%), and New Zealand (39%) were less inclined to start treatment than GPs in Germany (73%), Ireland (62%), and Switzerland (52%) (P = 0.05). Overall, GPs were less inclined to start treatment in 85-year-old than in 70-year-old females (pooled odds ratio [OR] 0.74 [95% confidence interval [CI] = 0.63 to 0.87]). Females with a TSH of 15 mU/L were more likely to get treated than those with a TSH of 6 mU/L (pooled OR 9.49 [95% CI = 5.81 to 15.5]).

Conclusion

GP treatment strategies of older people with subclinical hypothyroidism vary largely by country and patient characteristics. This variation underlines the need for a new generation of international guidelines based on the outcomes of randomised clinical trials set within primary care.     

Short term effects of infliximab on the lipid profile in patients with rheumatoid arthritis

OBJECTIVE: Cardiovascular morbidity and mortality appear to be enhanced in rheumatoid arthritis (RA), which might be due to an increased prevalence of cardiovascular risk factors such as dyslipidemia. It was recently shown that effective disease modifying antirheumatic drug treatment had a favorable influence on the lipid profile in patients with active RA. As infliximab markedly reduces disease activity in RA, we investigated the effects of infliximab on the lipid profile. METHODS: Infliximab was administered at baseline and at 2 and 6 weeks in patients with active RA. Total cholesterol and HDL-cholesterol concentrations were measured and their ratio, the atherogenic index (an important cardiovascular risk factor indicator), was assessed. RESULTS: Sixty-nine patients were enrolled. The Disease Activity Index score (DAS-28) was 5.9 (SD +/- 1.4) at baseline and decreased to 4.6 (+/- 1.4) after 2 weeks and further to 4.1 (+/- 1.5) after 6 weeks. Total cholesterol level was 5.2 mmol/l at baseline and increased to 5.7 mmol/l (p < 0.001) at 2 weeks, and was 5.6 mmol/l (p < 0.001 vs baseline) at Week 6. For HDL-cholesterol these values were 1.5, 1.6 (p < 0.001), and 1.6 mmol/l (p < 0.001 vs baseline), respectively. Changes in disease activity were significantly inversely associated with changes in total cholesterol and HDL-cholesterol levels. The atherogenic index, however, remained constant. Corticosteroid use at baseline was associated with significantly higher total cholesterol and HDL-cholesterol levels and a lower (more favorable) atherogenic index at baseline. CONCLUSION: Infliximab treatment was associated with a significant increase of both total cholesterol and HDL-cholesterol levels, which correlated with decreasing disease activity. However, this was not accompanied by a favorable effect on the atherogenic index. The favorable effect of infliximab on cardiovascular comorbidity might not be mediated by effects on lipid metabolism, but longterm investigations are needed to confirm this.     

Biosynthetic (recombinant) human granulocyte-macrophage colony- stimulating factor: effect on normal bone marrow and leukemia cell lines

To examine the biologic properties of the molecule encoded by the human gene for granulocyte-macrophage colony-stimulating factor (GM-CSF), we expressed the cloned complementary DNA (cDNA) in transfected monkey COS cells and purified the resultant protein. Purified biosynthetic human GM-CSF was added to cultures of normal hematopoietic progenitor cells in semisolid media, and the resulting colonies were characterized cytochemically. Non-adherent light-density bone marrow cells from healthy adult volunteers were maximally stimulated with GM-CSF (approximately 250 pmol/L, and four types of colonies were consistently identified by aspirating the individual colonies and staining with a triple stain for specific and nonspecific esterases and eosinophilic granules. Pure neutrophilic granulocyte (G), mixed granulocyte- macrophage (GM), pure macrophage (M), and pure eosinophil (EO) colonies were observed, the mean incidences on day 8 being 70%, 20%, 5%, and 5%, and on day 14, 7.5%, 16.6%, 50.9%, and 25.0%, respectively. In all types of colonies, complete maturation to segmented forms or typical macrophages was detected. GM-CSF did not enhance the growth of BFU-E from normal peripheral blood buffy coat cells in the simultaneous presence of erythropoietin alone or erythropoietin with purified erythroid-potentiating activity. GM-CSF stimulated HL-60 and KG-1 colony formation twofold and fivefold, respectively; consistent differentiation induction towards monocytic and eosinophilic lineages was observed in HL-60 but not in KG-1. These in vitro findings indicate that GM-CSF is a multilineage stimulator for progenitor cells of G, GM, M, and EO colonies.