人类免疫缺陷病毒/丙型肝炎病毒混合感染者抗丙型肝炎病毒治疗的进展

由于人类免疫缺陷病毒（HIV）和丙型肝炎病毒（HCV）有相同的传播途径,HIV/HCV混合感染现象十分普遍,已成为严重的公共卫生问题。高效抗逆转录病毒治疗（HAART）的应用显著减少了与HIV感染相关的发病率和病死率,而HCV混合感染引起的慢性肝脏疾病日益成为HIV/HCV混合感染者发病和死亡的重要因素。HIV/HCV混合感染者HCV相关肝病的风险增加,有效的抗HCV治疗对延长这一人群的生存期至关重要。本文就抗HCV治疗对象的评估、治疗时机的选择、治疗的方法、治疗监测和疗效评估以及治疗注意的问题作一综述。     

A rapid method for obtaining mesenchymal stem cells and platelets from bone marrow aspirate

Mesenchymal stem cells (MSCs) and platelet‐rich plasma (PRP) are currently used alone or in combination for therapeutic applications especially for bone repair. We tested whether MSCs can be isolated from bone marrow (BM) aspirate using a commercially available kit commonly used to obtain PRP from peripheral blood (PB). Results revealed that mononuclear cells and platelets from both PB and BM could be efficiently isolated by obtaining a mononuclear and platelet rich fraction (PB‐MPRF and BM‐MPRF, respectively). Starting with comparable volumes, the number of platelets increased 1.5‐fold in BM‐MPRF compared to PB‐MPRF. The number of clonogenic cells in BM‐MPRF samples was significantly higher than whole BM samples as revealed by CFU‐F assay (54.92 ± 8.55 CFU‐F/1.5 x 10⁵ nucleated cells and 32.50 ± 12.43 CFU‐F/1.5 x 10⁵ nucleated cells, respectively). Cells isolated from BM‐MPRF after in vitro expansion fulfilled the definition of MSCs by phenotypic criteria, and differentiated along osteogenic, adipogenic and chondrogenic lineages following induction. Results showed that the kit isolated MSCs and platelets from BM aspirate. Isolated MSCs were further expanded in a laboratory and BM‐MPRF was used clinically following BM withdrawal for rapid intra‐operative cell therapy for the treatment of bone defects. Copyright © 2012 John Wiley & Sons, Ltd.     

Human herpesvirus 8 enhances human immunodeficiency virus replication in acutely infected cells and induces reactivation in latently infected cells

Human herpesvirus 8 (HHV-8) is etiologically associated with Kaposi sarcoma (KS), the most common AIDS-associated malignancy. Previous results indicate that the HHV-8 viral transactivator ORF50 interacts synergistically with Tat protein in the transactivation of human immunodeficiency virus (HIV) long terminal repeat (LTR), leading to increased cell susceptibility to HIV infection. Here, we analyze the effect of HHV-8 infection on HIV replication in monocyte-macrophage and endothelial cells, as potential targets of coinfection. Primary or transformed monocytic and endothelial cells were infected with a cell-free HHV-8 inoculum and subsequently infected with lymphotropic or monocytotropic strains of HIV. The results show that HHV-8 coinfection markedly increases HIV replication in both cell types. HHV-8 infection induces also HIV reactivation in chronically infected cell lines and in peripheral blood mononuclear cells (PBMCs) from patients with asymptomatic HIV, suggesting the possibility that similar interactions might take place also in vivo. Furthermore, coinfection is not an essential condition, since contiguity of differently infected cells is sufficient for HIV reactivation. The results suggest that HHV-8 might be a cofactor for HIV progression and that HHV-8-infected endothelial cells might play a relevant role in transendothelial HIV spread.     

热休克蛋白27对人晶状体上皮细胞氧化损伤的保护作用及其机制

目的:观察热休克预处理诱导的热休克蛋白27对晶状体上皮细胞的保护作用。方法:实验于2004-01/12在中国医科大学卫生部小儿先天畸形重点实验室完成。体外培养人晶状体上皮细胞系HLB-C3细胞,随机分为4组:①正常对照组。②氧化损伤组:细胞中加入200μmol/LH2O2作用6h。③热休克处理组:42℃预热30min后,37℃恢复2h,处理同氧化损伤组。④放线菌酮组:预热前30min加入放线菌酮(25mg/L),其余处理同热休克处理组。观察各组细胞存活率、超氧化物歧化酶和过氧化氢酶活性;DNAladder和AnnexinⅤ-FITC流式细胞仪检测晶状体上皮细胞凋亡率;免疫印迹法检测各组中热休克蛋白27和半胱氨酸天冬氨酸蛋白酶3的表达。结果:①细胞活力:氧化损伤组低于正常对照组(0.24±0.02,0.28±0.04,P<0.01),热休克处理组高于氧化损伤组(0.27±0.04,P<0.05),放线菌酮组低于热休克处理组(0.23±0.01,P<0.01)。②细胞凋亡率:氧化损伤组高于正常对照组[(15.69±1.54)%,(4.17±0.83)%,P<0.01];热休克处理组低于氧化损伤组[(8.34±1.19)%,P<0.01];放线菌酮组高于热休克处理组[(13.58±1.21)%,P<0.01]。③超氧化物岐化酶和过氧化氢酶活性:氧化损伤组低于正常对照组(P<0.01),热休克处理组高于氧化损伤组(P<0.05),放线菌酮组低于热休克处理组(P<0.01)。④热休克蛋白27表达:正常对照组细胞中略有表达,氧化损伤组表达稍有增加,热休克处理组表达明显增多,放线菌酮组未见表达。⑤半胱氨酸天冬氨酸蛋白酶3的表达:正常对照组细胞表达较弱,氧化损伤组表达增多,热休克处理组的表达少于氧化损伤组。结论:热休克预处理诱导的热休克蛋白27对氧化损伤的晶状体上皮细胞起着保护作用。其保护机制可能是:①对与晶状体抗氧化有关的酶防御系统的影响:②小分子热休克蛋白对抗细胞凋亡。     

Atrial fibrillation recurrence after drug-induced typical atrial flutter ablation.

BACKGROUND: Catheter ablation of typical atrial flutter (AFL) occurring in patients who take antiarrhythmic drugs for atrial fibrillation (AF) has been proposed as a curative approach for AF. The aim of this study was to evaluate the efficacy of this technique. METHODS: Forty-six consecutive patients (30 males, 16 females, mean age 67 +/- 9 years) with paroxysmal or persistent AF were submitted to right atrial isthmus ablation: 1) 33 patients (group 1) in whom typical AFL spontaneously occurred during oral treatment with propafenone (n = 19), flecainide (n = 9) or amiodarone (n = 6); 2) 13 patients (group 2) submitted to electrophysiological study while taking oral propafenone (n = 3), flecainide (n = 8) or amiodarone (n = 1), in whom sustained AFL was induced (n = 9) or AF was induced and AFL was obtained by intravenous administration of class IC drugs (n = 4). The same antiarrhythmic drug which induced the conversion of AF into AFL was administered after ablation. RESULTS: During a follow-up of 20 +/- 18 months (range 1-78 months), 23 patients (50%) remained asymptomatic and free from AF recurrences. Fifteen patients (33%) with AF recurrences reported a reduction in arrhythmia-related symptoms. Eight patients (17%) did not show symptomatic improvement. These results did not significantly differ between group 1 and group 2. The duration of follow-up was significantly longer in patients with AF recurrence. Among several clinical, echocardiographic and electrophysiological parameters, only atrial enlargement and the absence of structural heart disease were independently associated with AF recurrence. CONCLUSIONS: In selected patients with AF and drug-induced AFL, right atrial isthmus ablation and prosecution of the drug treatment is a safe and feasible approach, which totally eliminates or reduces symptomatic AF recurrences in one half and one third of patients, respectively. However, the number of AF-free patients tends to decrease over time.     

Impact of physicians' beliefs and practices on cholesterol levels in patients with type 2 diabetes: a longitudinal assessment

Background

Clinical trials demonstrate significant benefit from cholesterol management for patients with type 2 diabetes. The aim of this work was to explore the correlates of lipid management in patients with type 2 diabetes, including the subjective beliefs of physicians, setting of care, and patient-related factors.

Methods

This longitudinal outcomes research study involved 2359 patients with type 2 diabetes recruited by 111 general practitioners and 214 physicians practicing in diabetes clinics. Physicians' beliefs were assessed through a questionnaire administered when the study started in 1998. Main outcome measures were total cholesterol (TC) and LDL cholesterol (LDL-C) levels over 3 years and the proportion of patients treated with lipid-lowering drugs (LLDs).

Results

Less than one-third of the physicians (27%) stated that they routinely started pharmacologic therapy for TC values ≥200 mg/dL (more aggressive), whereas 46% considered a TC level ≥240 mg/dL as the threshold for the initiation of treatment (less aggressive). During 3 years of observation, mean TC and LDL-C levels decreased from 215 ± 40 mg/dL to 203 ± 37 mg/dL and from 135 ± 36 mg/dL to 126 ± 35 mg/dL respectively, while the proportion of patients treated with LLDs increased from 13.2% to 24.6%; in particular, among individuals cared for by the more aggressive physicians, 30.0% were taking LLDs after 3 years, while only 17.7% of those followed by the less aggressive physicians and 18.1% of those followed by >1 physician were being treated with LLDs. Multilevel analysis showed that physicians' beliefs were an independent predictor of TC levels over the 3-year period. In patients treated with LLDs, TC levels decreased on average by 14%, and LDL-C levels decreased by 20%.

Conclusion

Our data show that physicians' beliefs in more aggressive management strategies will result in better mean TC values over a 3-year period.     

Extreme insulin resistance due to anti-insulin receptor antibodies: a direct demonstration of autoantibody secretion by peripheral lymphocytes

A 59-year-old woman with systemic lupus erythematosus was found to have marked hyperglycemia, extreme insulin resistance and abnormally high plasma immunoreactive insulin. Her circulating erythrocytes displayed a dramatic decrease of 125I-labeled insulin binding. Both the whole serum and purified IgG fraction strongly inhibited the binding of radiolabeled insulin to control erythrocytes. These results suggested, although indirectly, the existence of antibodies to insulin receptors in the serum of the patient. To directly investigate this issue, we used an enzyme-linked solid-phase immunoassay which allows the detection and enumeration of lymphocytes secreting antibodies towards insulin receptors. Peroxidase-conjugated anti-human immunoglobulin is used to reveal the binding of antibodies to insulin receptor-coated dishes. We demonstrated that the patient's mononuclear cells, when briefly incubated in Petri dishes with partially purified insulin receptor, were able to secrete immunoglobulins of G class specifically directed to the antigen. Moreover, only a fraction of the whole population of anti-insulin receptor antibodies was directed towards the insulin binding region of the receptor, seemingly corresponding to the auto-antibodies detected with conventional binding-inhibition assay.     

Endoscopic assessment of acute inflammation of the ileal reservoir after restorative ileo-anal anastomosis

Forty-seven patients, undergoing ileo-anal anastomosis for ulcerative colitis (42) or familial polyposis (5), were endoscopically examined after protective ileostomy or after restorative ileo-anal anastomosis. The neorectum and the ileum above were examined in all cases and multiple biopsies were taken. No symptoms of pouch inflammation were found in 41 subjects; 80.5% of these had non-macroscopic lesions and 19.5% had focal lesions such as congestion, petechiae, mucous hypersecretion (5), or single ulcers (3). None of these developed pouchitis. Pouchitis was observed in the other six subjects, who all underwent surgery for ulcerative colitis and developed 14 clinical episodes of pouchitis during the follow-up. In these cases the entire neorectum mucosa was always affected by the lesions which, in 50%, also extended to the ileum above. The most common endoscopic features (71.4%) were congestion, potechiae, oozing areas, mucous hypersecretion, and multiple superficial ulcers. In half the remaining cases (14.3%) the neorectum showed the features, described above, while the upper ileum was affected by deep round or irregular ulcers within normal mucosa; Crohn's disease was excluded in these cases. In the remaining 14.3%, pouchitis showed a pseudomembranous feature. In our experience, the endoscopic pattern of pouchitis is polymorphic. Although an ulcerative colitis-like feature prevails, pseudomembranous and Crohn's ileitis-like features may also be present.     

“The Linosa Study”: Epidemiological and heritability data of the metabolic syndrome in a Caucasian genetic isolate

Background and aimsGrowing evidence suggests that the metabolic syndrome (MetS) has both a genetic and environmental basis. To evaluate the possibility of a further genetic analysis, we estimated prevalence rates and heritabilities for the MetS and its individual traits in the adult population of Linosa, a small and isolated Italian Island in the southern-central part of the Mediterranean Sea.Methods and resultsThe Linosa Study (LiS) group consisted of 293 Caucasian native subjects from 51 families (123 parents; 170 offsprings). The MetS was defined according to NCEP/ATP III criteria and the following prevalence rates were calculated: hyperglycaemia 20.3%; central obesity 34.9%; hypertension 43.4%; hypertriglyceridaemia 29.9%; “low HDL” 56.6%; MetS 29.9%. Waist circumference was significantly related to all the quantitative parameters included in the NCEP/ATP III MetS definition. The MetS showed a heritability of 27% (p = 0.0012) and among its individual components, treated as continuous and discrete traits, heritability ranged from 10% for blood glucose to 54% for HDL-cholesterol. Among MetS subtypes, the clustering of central obesity, hypertriglyceridaemia and “Iow HDL” had the highest heritability (31%; p < 0.001).ConclusionThese data showed high prevalence rates for the MetS and its related traits in an isolated and small Caucasian population. The appreciable heritability estimates for the MetS and some of its components/clusters in the LiS population might support the observation of genetic factors underlying the pathogenesis of the MetS and encourage further analysis to identify new susceptibility genes.