Some metabolic changes during human corneal organ culture

In order to evaluate the changes in medium composition during closed system corneal organ culture at 30 degrees C, pH, pO2, glucose and lactate was measured 1 to 7, 14 and 28 days after incubation of a total of 49 human corneas. During organ culture pO2 decreased from 17.3 kPa to 8.8 kPa at day 28. The average oxygen consumption was initially 0.16 mumole/h/cm2, but decreased rapidly to an average of 0.04 mumole/h/cm2 during the first weeks of incubation. The glucose concentration fell from 5.1 mM to 0.25 mM after 28 days, and the lactate concentration rose from 2.5 mM to 11 mM. The concentration changes were for both substances highest during the first 14 days of organ culture. The pH fell from 7.36 to 6.64 after 28 days. These findings indicate that the cultured human cornea predominantly metabolizes glucose anaerobically, and that the metabolic activity decreases during incubation. The limiting metabolic factors for prolonging the culture period seem to be development of severe acidosis and glucose depletion in the medium.     

Neutrophil chemotactic factors derived from conjunctival epithelial cells: preliminary biochemical characterization

We have previously reported on the release of neutrophil chemotactic factors (NCF) from injured conjunctival tissue. The present study was designed to biochemically characterize these conjunctiva-derived chemotactic factors and determine their biological activities. Bulbar conjunctiva was surgically removed from a rabbit eye and incubated with 250 microL of minimal essential medium (MEM) for 6 hours at 37 degrees C in a 5% CO2 atmosphere. Chemotactic activity was assayed using modified Boyden chambers with rabbit peritoneal neutrophils as indicator cells. Following treatment with subtilisin protease for 90 minutes, chemotactic activity of the conjunctival factors was reduced by 74%. Similarly, activity was lost after heating at 56 degrees C for 60 minutes (41% inhibition). Using ultrafiltration techniques, we showed that the majority of the chemotactic activity remained above a 100 kilodalton filter, suggesting the existence of high molecular weight factors. We also showed that the conjunctival factors are not glycoproteins and bind to both anion and cation exchange resins. When 100 microL of conjunctival supernatant was injected in the superior tarsal conjunctiva of rabbits, significant recruitment of neutrophils was evident by 4 hours. Control rabbits injected with MEM did not show neutrophil recruitment. Results of these studies indicate that NCF from traumatized conjunctival tissue are proteins (and not glycoproteins) of high molecular weight, heat labile, exhibit anionic and cationic charges, and are active in vivo.     

Surreptitious drug use by patients in a panic disorder study

In a double-blind, placebo-controlled trial comparing alprazolam and imipramine for panic disorder, serum analysis revealed that a substantial proportion of the patients took explicitly prohibited anxiolytic medication. Excluding these patients changed the results.     

The social function of the hospital

Issuing from the statements to the change of the social institution hospital in the course of history the experiment is undertaken to outline the social function of the hospital on the conditions of the developed socialist society.     

NBS1 expression as a prognostic marker in uveal melanoma.

PURPOSE: Up to half of uveal melanoma patients die of metastatic disease. Treatment of the primary eye tumor does not improve survival in high-risk patients due to occult micrometastatic disease, which is present at the time of eye tumor diagnosis but is not detected and treated until months to years later. Here, we use microarray gene expression data to identify a new prognostic marker. EXPERIMENTAL DESIGN: Microarray gene expression profiles were analyzed in 25 primary uveal melanomas. Tumors were ranked by support vector machine (SVM) and by cytologic severity. Nbs1 protein expression was assessed by quantitative immunohistochemistry in 49 primary uveal melanomas. Survival was assessed using Kaplan-Meier life-table analysis. RESULTS: Expression of the Nijmegen breakage syndrome (NBS1) gene correlated strongly with SVM and cytologic tumor rankings (P < 0.0001). Further, immunohistochemistry expression of the Nbs1 protein correlated strongly with both SVM and cytologic rankings (P < 0.0001). The 6-year actuarial survival was 100% in patients with low immunohistochemistry expression of Nbs1 and 22% in those with high Nbs1 expression (P = 0.01). CONCLUSIONS: NBS1 is a strong predictor of uveal melanoma survival and potentially could be used as a clinical marker for guiding clinical management.     

Treatment of central corneal ulcers by à chaud transplantation of organ culture preserved donor tissue

In the period 1979-83 a total of 232 penetrating corneal transplantations were performed with organ culture preserved donor material. Forty-two operations were made à chaud for central corneal ulcerations. The results of these interventions are reported and discussed. The surgery presented no serious problems and on first post-operative day the anterior chamber was reformed in all cases, and pain was relieved. Patients were confined to bed only for 1 day. Graft deswelling was slightly retarded compared to planned surgery, but all functioning grafts obtained normal thickness. The overall graft survival rates after 6, 12 and 18 months were 66, 53 and 48%, respectively. For herpetic ulcers the survival after 18 months was 49% and for non-herpetic ulcers 47%. The grafts failed in all cases where it had been necessary to employ grafts of more than 10 mm. This latter group was comprised of the non-herpetic cases; were they excluded, the graft survival rate would increase to 60%. Among cases with a clear graft the median visual acuity after 18 months was 0.33 (range less than 0.1-0.67). Excluding cases with a non-corneal cause of visual reduction, the median visual acuity was 0.4 (range 0.33-0.67). It is concluded that grafting can be successfully employed in the treatment of central ulcers, as it not only restitutes tissue integrity, but also preserves useful vision.     

骨形态发生蛋白2表达异常与脊柱的融合

目的:综述近几年来国内外对骨形态发生蛋白2表达异常及其基因突变的研究,重点分析突变对脊柱融合的影响,为基因重组骨移植提供理论依据。资料来源:应用计算机检索Medline和Science Direct Online数据库1989-01/2007-01期间与骨形态发生蛋白2表达异常及脊柱融合相关的文章,检索词为"BMP2,BMP,gene mutation,mutation,gene expression,abnormal expression,spinal fusion,bony fusion,bone transplantation",限定文章语言种类为English。同时计算机检索中国期刊全文数据库2000-01/2007-01期间与骨形态发生蛋白2表达异常及脊柱融合相关的文章,检索词为"骨形态发生蛋白2,脊柱融合,骨移植,基因突变,表达异常",限定文章语言种类为中文。资料选择:对资料进行初审,选择有关骨形态发生蛋白2生物学活性、信号转导机制、基因突变、与骨诱导的关系、在脊柱融合中的研究进展的文献,共收集到121篇,排除综述类及重复研究。资料提炼:选择其中有代表性的30篇进行综述,涉及到骨形态发生蛋白2的生物学活性、诱导成骨机制、突变的分子学基础、基因突变及影响蛋白表达异常的其他因素。资料综合:骨移植植骨融合的形成是一个多因子、多基因的过程,涉及到骨生成、骨诱导和骨传导3个环节。骨形态发生蛋白2是骨发育和修复的关键调节剂,骨折愈合时尤其需要。国内外学者对于骨形态发生蛋白2基因的自身突变研究取得了很大进展,已检测出不同部位多个位点的突变。基因变异导致所编码氨基酸发生改变,从而引起相应多肽结构发生变异,影响蛋白质的理化性质。无论是体内的骨形态发生蛋白2抑制因子,突变导致的生物活性改变,还是其增龄性效应,最终都将引起局部骨形态发生蛋白2含量下降及生物学活性减退,可能直接导致脊柱融合术后植骨愈合不良或植骨不愈合的发生。结论:对骨形态发生蛋白2表达异常尤其对其基因进行突变分析,寻找影响植骨融合的遗传学因素,具有非常重要的临床意义。     

Sensitivity and specificity of four assays to detect human T− lymphotropic virus type I or type I/II antibodies

BACKGROUND: Assays that detect human T-lymphotropic virus type I and type II antibody (HTLV-I/II) are widely used in the routine screening of blood donors. STUDY DESIGN AND METHODS: Four commercially available anti-HTLV-I (Fujirebio and Organon Teknika) or -HTLV-I/II assays (Murex and Ortho) were evaluated in various serum panels: A) HTLV-I-positive specimens (n = 41), confirmed by Western blot and polymerase chain reaction; B) a commercially available anti-HTLV-I/II panel; C) serial dilutions of sera from HTLV-I-positive individuals (n = 30), confirmed by immunofluorescence assay and Western blot: D) serial dilutions of HTLV-II-positive blood donors (n = 20), confirmed by Western blot and polymerase chain reaction, and E) sera from first-time blood donors (n = 1055). RESULTS: All four assays elicited reactions in all 82 HTLV-I- positive samples in Panels A, B, and C. Of 32 HTLV-II-positive specimens in Panels B and D, 31 (96.9%) reacted in the Organon Teknika assay and all 32 reacted in the remaining tests. Probit analysis of test results in Panels C and D indicated that the Fujirebio test was the most sensitive assay, followed by Organon Teknika, Ortho, and Murex. The specificities of Fujirebio, Murex, Organon Teknika, and Ortho tests in 1055 first-time blood donors were 99.9, 100, 99.6, and 99.9 percent, respectively. CONCLUSION: All four studied assays for detecting HTLV-I or HTLV-I/II antibodies are appropriate as screening tests.     

紫杉醇药物涂层支架植入术后患者外周血单核细胞中热休克蛋白70的变化：生物材料临床应用近期效果随访

目的:植入材料、靶血管病变特征、术前状态、炎症因子及急性期蛋白均对急性冠状动脉综合征接受支架材料介入治疗后的效果有影响,为验证紫杉醇涂层支架临床应用后材料及宿主的相关反应,实验观察了接受紫杉醇涂层支架介入治疗的急性冠状动脉综合征患者的外周血热休克蛋白70水平变化,并分析其临床意义。方法:①连续性入选2004-12/2006-03在江苏大学附属人民医院行经皮冠状动脉介入治疗的78例急性冠状动脉综合征患者,全部病例均置入紫杉醇药物涂层支架。采用流式细胞仪测定症状发作平均(34.1±16.2)h的外周血单核细胞热休克蛋白70阳性表达水平。②所有患者随访至术后6个月,出现心源性死亡、再次心肌梗死、再发心绞痛、再次血运重建术和继发心衰等主要心脏不良事件者为近期预后不良组,无上述情况者判定为近期预后良好组,用logistic多元回归法分析术前状态、靶血管病变特征、植入支架的各项参数及外周血热休克蛋白70水平与主要心脏不良事件发生率的关系,并以同期健康体检者20例为正常对照组。结果:68例患者完成随访进入结果分析。①外周血热休克蛋白70水平:急性心肌梗死患者和不稳定型心绞痛患者比较差异无统计学意义(P>0.05),但均显著高于正常对照组(P<0.05)。②在多变量的logistic回归分析中,外周血热休克蛋白70独立于其危险因素,能预测急性冠状动脉综合征患者经皮冠状动脉介入治疗后近期主要心脏不良事件发生率(OR值为0.904,P<0.05)。结论:回归分析结果提示,应用紫杉醇涂层支架临床治疗近期效果评估中,外周血热休克蛋白70水平高的急性冠状动脉综合征患者近期心脏事件发生率较高,说明外周血热休克蛋白70可能成为判断紫杉醇涂层支架介入治疗后不良事件发生率的独立因素之一。