糖尿病大鼠骨代谢变化与立普妥、胰岛素的干预效应

目的:近年细胞培养实验发现他汀类药物可以促进骨形成,采用动物实验观察胰岛素和他汀类药物立普妥对糖尿病大鼠骨代谢的影响,为糖尿病伴骨质疏松的治疗提供实验依据。方法:实验于2005-08/2006-01在大连医科大学病理教研室完成。①实验分组:SD雄性大鼠55只,随机选择10只为空白对照组,余45只经鼠尾静脉注射链尿佐菌素造成糖尿病大鼠模型。其中40只符合造模标准,随机分为糖尿病未治疗组、胰岛素治疗组、立普妥治疗组及胰岛素 立普妥治疗组,每组10只。②实验方法:所有大鼠皆给予相同普通饮食。胰岛素治疗组及胰岛素 立普妥治疗组于实验第4天接受中效胰岛素治疗,6～8U/d分两次颈背部皮下注射。胰岛素剂量按每只鼠每周血糖进行调整。立普妥治疗组及胰岛素 立普妥治疗组于实验第4天给予立普妥1.25mg/kg灌胃。糖尿病未治疗组和空白对照组给予等量生理盐水灌胃。③实验评估:9周末用乙醚麻醉,每组取4只大鼠去眼球取血之后处死。14周末应用同样方法处死剩余大鼠。均取腰椎骨,常规脱钙石蜡包埋,行苏木精-伊红染色。骨组织形态计量学测量平均骨小梁厚度和平均骨小梁间距或弥散度。血中Ⅰ型胶原氨基端肽测定采用竞争性放射免疫检测方法(碘标记)。结果:实验期间大鼠死亡5只,其中糖尿病未治疗组1只于第3周死亡,胰岛素组2只于第6周死亡,胰岛素 立普妥治疗组2只于第7周死亡。①骨组织病理形态学变化:9周末立普妥治疗组、胰岛素 立普妥治疗组及糖尿病未治疗组光镜下见骨质疏松表现。14周末立普妥治疗组及胰岛素治疗组骨组织微观结构恢复至空白对照组水平。②平均骨小梁厚度:9周末:糖尿病未治疗组、胰岛素治疗组、立普妥治疗组及胰岛素 立普妥治疗组均明显低于空白对照组(P<0.01)。14周末:糖尿病未治疗组及胰岛素 立普妥治疗组均明显低于空白对照组(P<0.05)。③平均骨小梁间距或弥散度:9周末:糖尿病未治疗组及胰岛素 立普妥治疗组均明显高于空白对照组(P<0.05)。14周末:糖尿病未治疗组及胰岛素 立普妥治疗组均明显高于空白对照组(P<0.05)。④Ⅰ型胶原氨基端肽水平:9周末:立普妥治疗组和胰岛素 立普妥治疗组均明显高于空白对照组(P<0.01)。14周末:胰岛素治疗组、立普妥治疗组和胰岛素 立普妥治疗组均明显高于空白对照组(P<0.01)。结论:①糖尿病大鼠造模9周出现明显的骨质疏松。②糖尿病大鼠骨质变化表现为骨吸收超过骨形成作用,主要以骨吸收增强为主。③立普妥及胰岛素可以促进糖尿病大鼠骨质的形成,抑制糖尿病大鼠骨质疏松的发生发展。     

First molecular evidence of Tula hantavirus in Microtus voles in Slovenia

Different Microtus species, present in a worldwide range habitat populating North America, Europe, Asia, and few other species have been recognized previously as a hantavirus reservoir. Tula hantavirus was first reported in Microtus arvalis and Microtus rossiaemeridionalis from Central Russia and later discovered in several European countries. Using molecular techniques we have demonstrated the presence of Tula hantavirus in three different Microtus species in Slovenia. Phylogenetic analyses of partial S segment placed Slovenian strains in the same genetic lineage as Austrian and Croatian strains.     

Nucleotide sequences of novel HLA-B35 subtypes

In recent reports we described novel hybridization patterns (HP) corresponding to 22 potentially new HLA-B locus alleles in a panel of 547 subjects studied by PCR-SSOP. Three of them correspond to new subtypes of B35. To confirm the hybridization results we have isolated DNA from PBL and performed PCR, DNA cloning and nucleotide sequencing. One of the alleles, locally called B-3505v was found in three individuals: two Hispanic, one Caucasoid. It differs from B^*3505 by 3 nucleotide substitutions that lead to changes in residues 94 (Ile > Thr), 95 (Ile > Leu) and 103 (Val > Leu). B-3505v differs from B^*3501 in residues 97 and 103. Another allele called B-3508v, was found in 7 individuals, (6 of 122 Toba Indians, 1 of 18 Pilaga Indians). It differs from B^*3509 in two silent nucleotide substitutions (codons 135 and 138) and in one substitution in residue 156 (Arg > Leu). The new allele has a hybrid sequence between B^* 3508 and B^*4801. A third subtype, locally called B-3504v, observed in two Hispanic individuals, is identical to B^*3512. B^*3512 differs from B^*3504 by 3 nucleotides and one amino acid. Substitutions in residue 95 contribute to the structure of specificity pocket F, 97 to C and E, and 156 to pockets D and E. Therefore it is possible that some of the new alleles may have different peptide binding profiles. Since differences in residue 156 have been shown to affect allorecognition and mediate GvHD, identification of such variants may have important implications in transplantation and perhaps in studies of immune responses to peptides and pathogens.     

青少年特发性脊柱侧凸主弯侧凸Cobb角、胸椎后凸角及腰椎前凸角仰卧位MRI与站立位X线测量的对比研究

目的 探讨仰卧位MRI与站立位X线全脊柱片在青少年特发性脊柱侧凸(AIS)患者侧凸Cobb角、胸椎后凸角(TK)及腰椎前凸角(LL)测量上的差异性和相关性。方法 选取2008年1月—2016年1月行外科支具治疗或手术治疗且密切随访的120例AIS患者的站立位X线全脊柱片和仰卧位MRI全脊柱重建片资料进行回顾性分析。于站立位X线全脊柱正侧位片上分别测量主弯侧凸Cobb角、TK及LL。于患者仰卧位MRI全脊柱冠状面和矢状面重建片上的相同节段测量侧凸Cobb角、TK和LL。应用配对t检验及线性回归分析两组之间测量值的差异性及相关性。结果 120例站立位X线片上和仰卧位MRI上主弯侧凸Cobb角分别为33.8°±20.1°和24.9°±18.3°,TK分别为15.2°±9.7°和10.5°±7.7°,LL分别为43.6°±13.8°和37.1°±13.5°,差异均有统计学意义(P值均<0.01)。线性相关分析显示,站立位X线片上和仰卧位MRI上主弯侧凸Cobb角、TK和LL均有相关性,r分别为0.920、0.706和0.565(P值均<0.01)。线性回归分析得回归方程为:＾Y_{站立位X线侧凸Cobb角}=0.901×X_{仰卧位MRI侧凸Cobb角} +12.517、＾Y_{站立位X线TK}=1.055×X_{仰卧位MRI TK}+3.865、＾Y_{站立位X线LL}=0.718×X_{仰卧位MRI LL}+17.135。结论 仰卧位MRI与站立位X线片在AIS患者主弯侧凸Cobb角、TK及LL的测量具有差异且呈线性相关,利用仰卧位MRI可计算出站立位X线片上患者的侧凸Cobb角、TK和LL。     

Diversity of ace, a gene encoding a microbial surface component recognizing adhesive matrix molecules, from different strains of Enterococcus faecalis and evidence for production of ace during human infections

Our previous work reported that most Enterococcus faecalis strains adhered to the extracellular matrix proteins collagen types I and IV and laminin after growth at 46 degrees C, but not 37 degrees C, and we subsequently identified an E. faecalis sequence, ace, that encodes a bacterial adhesin similar to the collagen binding protein Cna of Staphylococcus aureus. In this study, we examined the diversity of E. faecalis-specific ace gene sequences among different isolates obtained from various geographic regions as well as from various clinical sources. A comparison of nucleotide and deduced amino acid sequences of Ace from nine E. faecalis strains identified a highly conserved N-terminal A domain, followed by a variable B domain which contains two to five repeats of 47 amino acids in tandem array, preceded by a 20-amino-acid partial repeat. Using 17 other strains collected worldwide, the 5' region of ace that encodes the A domain was sequenced, and these sequences showed > or =97.5% identity. Among the previously reported five amino acids critical for collagen binding by Cna of S. aureus, four were found to be identical in Ace from all strains tested. Polyclonal immune rabbit serum prepared against recombinant Ace A derived from E. faecalis strain OG1RF detected Ace in mutanolysin extracts of seven of nine E. faecalis strains after growth at 46 degrees C; Ace was detected in four different molecular sizes that correspond to the variation in the B repeat region. To determine if there was any evidence to indicate that Ace might be produced under physiological conditions, we quantitatively assayed sera collected from patients with enterococcal infections for the presence of anti-Ace A antibodies. Ninety percent of sera (19 of 21) from patients with E. faecalis endocarditis showed reactivity with titers from 1:32 to >1:1,024; the only 2 sera which lacked antibodies to Ace A had considerably lower titers of antibodies to other E. faecalis antigens as well. Human-derived, anti-Ace A immunoglobulins G purified from an E. faecalis endocarditis patient serum inhibited adherence of 46 degrees C-grown E. faecalis OG1RF to collagen types I and IV and laminin. In conclusion, these results show that ace is highly conserved among isolates of E. faecalis, with at least four variants related to the differences in the B domain, is expressed by different strains during infection in humans, and human-derived antibodies can block adherence to these extracellular matrix proteins.     

Association between intraocular pressure and budesonide inhalation therapy in asthmatic patients.

BACKGROUND: The extent to which inhaled glucocorticoids increase the risk of intraocular pressure elevation has been controversial. OBJECTIVE: The authors attempt to assess such risk attributable to budesonide, an inhaled glucocorticoid for asthma therapy. METHODS: Data were pooled from four prospective, randomized, double-blind, parallel-group, placebo-controlled, multicenter clinical trials of 12 to 20 weeks in duration. One thousand two hundred and fifty-five patients, 6 to 70 years of age whose intraocular pressures (IOPs) were less than 23 mmHg at screening were randomized to receive placebo or inhaled budesonide at doses ranging from 100 to 800 microg, administered twice daily. Intraocular pressure was measured at screening and at the end of double-blind treatment. Intraocular change was compared between budesonide and placebo, accounting for the confounding effects of gender, race, age, history of diabetes, history of hypertension, clinical trial, systemic glucocorticoid use during the trials, ophthalmic glucocorticoid use during the trials, and prior oral glucocorticoid use. RESULTS: No budesonide treatment effect on the IOP was evident either in the crude analysis or after adjustment for possible confounding factors. For patients exposed to budesonide at a total daily dose of 1600 microg for 20 weeks, there was no difference in IOP change compared with the placebo controls. CONCLUSIONS: No association with an increased IOP was observed in asthmatic patients treated with budesonide at daily doses ranging from 200 to 1600 microg for durations of 12 to 20 weeks. The subgroup analysis, which focused on the highest dose and longer term therapy was reassuring, as was the overall result.     

Proton pump inhibitor and clopidogrel interaction: The case for watchful waiting

Clopidogrel is an integral part of the management of several important vascular diseases. However the medium to long term clinical outcomes are poorer for these patients if they experience gastro‐intestinal bleeding, hence patients with risk factors for gastro‐intestinal bleeding are frequently prescribed proton pump inhibitors. Conflicting evidence exists as to the existence of an adverse interaction between clopidogrel and proton pump. This review examines the original studies, which suggested the adverse interaction, the subsequent and most recent studies, the pharmaco‐dynamics of the two drugs and suggests an algorithm for the use of clopidogrel with proton pump inhibitors.     

Molecular typing of selected Enterococcus faecalis isolates: pilot study using multilocus sequence typing and pulsed-field gel electrophoresis

The present study compared the recently developed multilocus sequence typing (MLST) approach with a well-established molecular typing technique, pulsed-field gel electrophoresis (PFGE), for subspecies differentiation of Enterococcus faecalis isolates. We sequenced intragenic regions of three E. faecalis antigen-encoding genes (ace, encoding a collagen and laminin adhesin; efaA, encoding an endocarditis antigen; and salA, encoding a cell wall associated antigen) and one housekeeping gene (pyrC) of 22 E. faecalis isolates chosen largely for their temporal and geographical diversity, but also including some outbreak isolates. MLST analysis of polymorphic regions of these four genes identified 13 distinct sequence types (STs) with different allelic profiles; the composite sequences generated from the four sequenced gene fragments of individual isolates showed 98.3 to 100% identity among the 22 isolates. We also found that the allelic profiles from two sequences, ace and salA, were sufficient to distinguish all 13 STs of this study. The 13 STs corresponded to 12 different PFGE types, with one previously designated PFGE clone (a widespread U.S. clone of beta-lactamase-producing isolates) being classified into two highly related STs which differed at 2 of 2,894 bases, both in the same allele. MLST also confirmed the clonal relationships among the isolates of two other PFGE clonal groups, including vancomycin resistant isolates. Thus, this pilot study with representative E. faecalis isolates suggests that, similar to PFGE, the sequence-based typing method may be useful for differentiating isolates of E. faecalis to the subspecies level in addition to identifying outbreak isolates.     

Relationship of Quality of Life with Disability Grade in Obsessive Compulsive Disorder and Dysthymic Disorder

Background:

There is paucity of information on the relationship of quality of life (QOL) in obsessive compulsive disorder (OCD) and dysthymic disorder (DD) with disability grade in India.

Aim:

To assess the relation of QOL with disability level in OCD and DD.

Materials and Methods:

This hospital based study was conducted in a medical institution in Davanagere, Karnataka, India. Data was collected by using Diagnostic and Statistical Manual IV Text Revision (DSM IV TR) criteria, WHO QOL BREF and IDEAS. Relationship between disability grade and QOL was assessed by independent sample t test.

Results:

Mild disabled OCD patients had a significantly better QOL in the Q1 domain i.e. perception on quality of life as compared to moderately disabled patients (P < 0.05), while in other domains of QOL, there was no statistically significant difference (P > 0.05). But, QOL score in physical domain showed significant difference across disability grades (56.00, SD = 6.89; 48.50, SD = 12.28) in DD, but not in other domains.

Conclusion:

Perception of QOL is better in those with mild disability in OCD, but in DD, physical domain of QOL score is more in mild disability compared to moderate disability.