Influence of tissue preparation on the acoustic properties of tissue sections at high frequencies

The purpose of the present study was to clarify the influence of tissue preparation on the high-frequency acoustic properties by comparing the acoustic properties of the formalin-fixed, paraffin-embedded, deparaffinized sections and formalin-fixed, frozen sections for two types of fat-containing renal cancer and fat-free renal oncocytoma using a SAM. There was no significant difference for the sound speed among the clear cell, granular cell renal cancer and oncocytoma in either groups, but the attenuation constant was significantly higher for the frozen than for that of the paraffin section in fat-containing renal cancer. In fat-free oncocytoma, there was no significant difference for the attenuation constant in either group. The data suggest that the fat component, which had been eluted by paraffinization, is stored and the true acoustic properties of the tissue can be measured in frozen section.     

Effectiveness of electroacupuncture and interferential eloctrotherapy in the management of frozen shoulder.

OBJECTIVE: To examine whether the addition of either electroacupuncture or interferential electrotherapy to shoulder exercises would be more effective in the management of frozen shoulder. DESIGN: A double-blinded, randomized, controlled trial. METHODS: A total of 70 subjects were randomly allocated to receive either: (i) electroacupuncture plus exercise; (ii) interferential electrotherapy plus exercise; or (iii) no treatment (the control group). Subjects in groups (i) and (ii) received 10 sessions of the respective treatment, while the control group received no treatment for 4 weeks. Each subject's score on the Constant Murley Assessment and visual analogue scale were recorded at baseline, post-treatment session and subsequent follow-up sessions. RESULTS: In both the electroacupuncture and interferential electrotherapy groups, the Constant Murley Assessment score increased and the visual analogue scale score decreased significantly (both p < 0.001). No significant change was found in any outcome of the control group, and no significant difference was found between the 2 intervention groups (all p > 0.05). The observed improvement was well maintained in both intervention groups at least until the 6-month follow-up session. CONCLUSION: Either electroacupuncture or interferential electrotherapy in combination with shoulder exercises is effective in treating frozen shoulder patients. However, no significant difference was found between these types of treatment.     

Age-related changes in male forearm skin-to-fat tissue dielectric constant at 300 MHz

Prior research suggests that tissue dielectric constant (TDC) values are useful to assess localized skin water in females for early diagnosing breast cancer treatment-related lymphoedema and TDC values in young adults have shown gender differences. However, no TDC data are available for older males nor have ageing effects been studied despite known shifts in water state and other skin age-related changes. Thus our goals were to (i) characterize TDC values at various skin depths in young and older males, (ii) determine the dependence of these values on body composition parameters and (iii) establish inter-arm TDC ratios for use as normal male reference values. TDC measurements were made to depths of 0·5, 1·5, 2·5 and 5·0 mm bilaterally on volar forearm skin in 60 males in three groups of 20 that had mean ages ± SD of 24·0 ± 0·9, 40·0 ± 12·9 and 71·0 ± 8·0 years. Total body fat and water percentages were determined via bioimpedance at 50 KHz. Results showed that (i) for all age groups TDC values decreased with increasing depth, (ii) TDC values were not statistically different among age groups except at a depth of 0·5 mm, (iii) TDC values were highly negatively correlated with total body fat and (iv) inter-arm ratios varied little among age groups and depths. It is concluded that (i) age-related larger TDC values at only the shallowest depth is consistent with skin water shifting state from bound to more mobile in the oldest group and (ii) inter-arm ratios at any depth provide a basis to test for unilateral oedema.     

声触诊组织量化技术评估慢性肾病的影响因素及其应用价值

目的 研究声触诊组织量化（VTQ）技术评估慢性肾病的影响因素,评价其应用价值。方法 应用VTQ技术测量192例慢性肾病患者（病例组）及112名正常对照的肾皮质硬度,于右肾下极设置ROI,将检查深度设定为2.0~6.5 cm,并分为2.0~3.5 cm（深度1）、3.6~5.0 cm（深度2）、5.1~6.5 cm（深度3）,分析不同深度下各期肾病患者与正常对照组的剪切波速度（SWV）差异。评价患者的血肌酐与肾皮质SWV的相关性,分析不同病理类型间SWV差异。结果 深度1~3下,正常肾皮质的SWV中位数分别为2.91、2.82、2.48 m/s,深度1、深度2与深度3的SWV差异有统计学意义。深度2下,正常对照组与中晚期肾病组SWV差异有统计学意义,且随病变程度加重,肾皮质SWV中位数逐渐减小。肾病组肾皮质SWV值与血肌酐水平呈负相关。不同病理类型肾病间SWV差异无统计学意义。结论 VTQ技术可用于评估慢性肾病的病变程度,定量评价肾脏顺应性。     

下调miR-126对大鼠降血压作用的实验研究

目的探讨原发性高血压中miR-126是否可作为一种新的治疗靶点。方法 24只自发性高血压大鼠(SHR)随机分为4组,每周测量收缩压、舒张压、平均动脉压及心率。经过8周的病毒载体转染的处理,心、肾、脑、肝、肺组织被制成冰冻切片,通过荧光显微镜观察慢病毒载体在各个组织中的分布。应用ELISA试剂盒来测定血清NO含量。结果大鼠中下调miR-126无明显降血压的作用。结论通过下调miR-126来治疗原发性高血压并没有显著降血压的作用。高血压患者中miR-126表达的增加可能是由于一种代偿机制,具体机制有待进一步的研究。     

结肠癌组织中p16和CDK4的表达及相互关系

采用免疫组化法分析６０例结肠癌、结肠息肉和结肠炎标本中ｐ１６和ＣＤＫ４的蛋白表达。结肠癌和结肠息肉组织中ｐ１６表达随细胞分化程度降低而减少；结肠炎组织ｐ１６表达高于癌和息肉组，ＣＤＫ４在上述三种组织中表达无显著差异；推测结肠组织从癌前病变至癌变发展中有ｐ１６对ＣＤＫ４抑制失调。该检测方法亦为临床上区分癌与癌前病变及变化趋势提供辅助诊断方法。     

肝肾灰阶比值分析肝纤维化严重程度的临床价值

目的 探讨肝实质与肾皮质灰阶比值评定肝纤维化严重程度的临床应用价值.方法 常规腹部超声检查60例肝纤维化患者及40例健康志愿者;将超声仪器条件设置一致后,选取肝肾切面冻结后,将图像存储于超声图文工作站硬盘,图像导人计算机后利用Photoshop软件定制直方图取样范围,测量肝近程、中程、远程灰阶值及右肾皮质灰阶值,将肝实质与自身肾皮质的灰阶比值作为诊断肝纤维化的新指标,与病理结果进行统计学分析.结果 随肝纤维化程度加重,肝回声增强,而其自身肾皮质回声无明显变化,肝肾灰阶比值相应增加,与预测结果一致;肝纤维化组与对照组间肝肾灰阶比值差异有统计学意义(P＜0.01).结论 超声测定的肝肾灰阶比值较少受患者个体透声状况等因素影响,可作为临床无创诊断肝纤维化、评价肝纤维化程度及观察其疗效的新指标.     

Immunoglobulin heavy-chain gene rearrangement studies by Southern blot using DNA extracted from formalin-fixed, paraffin-embedded tissue.

BACKGROUND: In most clinical molecular diagnostics laboratories, Southern blots for gene rearrangement studies are not routinely performed on formalin-fixed, paraffin-embedded (FFPE) tissue samples. In this study, immunoglobulin heavy-chain gene rearrangements by Southern blot using DNA extracted from FFPE tissue samples were studied. METHODS AND RESULTS: Eleven paired freshly frozen and FFPE tissue samples were evaluated for immunoglobulin gene rearrangements by PCR and Southern blot analyses. An additional 14 selected samples sent to our laboratory to rule out lymphoma, for which only FFPE tissue (no frozen tissue) was available and for which PCR was interpreted as negative, were evaluated by the same techniques. Southern blots generated from DNA extracted from FFPE tissues were qualitatively identical to those generated from DNA extracted from fresh or freshly frozen tissue and correlated well with the final diagnoses. Ten interpretable Southern blots were generated in the 14 cases in which no frozen tissue was available. Four of these ten blots were interpreted as positive for an immunoglobulin gene rearrangement. Although the number of samples analyzed is small, success with Southern blotting correlated with increased sample size and sample width (1.17 vs 0.49 cm(2); P <.024; 0.71 vs 0.43 cm; P <. 049, respectively). CONCLUSION: DNA extracted from FFPE tissue samples using the simple, efficient, and nontoxic techniques described in this report can be used in many cases for Southern blotting for the detection of clonality by gene rearrangement studies.     

不同冷冻方案保存人类卵巢组织的活性比较

背景：卵巢组织冷冻保存被认为是保存女性生殖内分泌功能安全、有效的方法,但目前尚无统一确定的方案。 目的：探讨3种冷冻方案对人类卵巢组织保存冷冻效果及卵泡活性的影响。方法：采用丙二醇慢速程序冷冻法、二甲基亚砜玻璃化冷冻法、液氮直投法对20例人类卵巢组织进行冷冻保存,复苏后采用细胞存活/死亡荧光分析法计数有活性的细胞,体外培养测定培养液雌二醇浓度及各级卵泡计数,判断3种不同冷冻方案对人类卵巢组织活性的影响。结果与结论：不同冷冻方案冻融后卵巢组织块的活性卵泡率均低于新鲜卵巢组（P 〈 0.05）,二甲基亚砜组最低,液氮直投法组与慢速程序冷冻法组差异无显著性意义。体外培养各冷冻组分泌的雌二醇水平比较,第4天时二甲基亚砜组低于液氮直投法组和慢速程序冷冻法组（P 〈 0.05）,至第8天时各冷冻组雌二醇水平与新鲜卵巢组一致。培养14 d组织学观察,各组卵巢组织内生长期卵泡比例增多,始基卵泡仍然占最主要的。新鲜卵巢组织正常卵泡的总数高于冷冻组（P 〈 0.05）,二甲基亚砜组的正常卵泡数低于液氮直投法组和慢速程序冷冻法组（P 〈 0.05）。提示冷冻保存对卵巢组织卵泡有一定的损伤,但仍能保存大部分始基卵泡的活性,经体外培养后可进一步发育并具有分泌功能,在3种冷冻方案中,慢速程序冷冻法和液氮直投法的冷冻效果优于二甲基亚砜玻璃化冷冻法,液氮直投法操作简便,冷冻效果稳定。     

The potential of amniotic fluid stem cells for cellular therapy and tissue engineering

INTRODUCTION: Foetal cells present in amniotic fluid (AF) have been used for many years to perform prenatal genetic screening. Recent reports suggested that these cells might have additional benefits. AF contains, in addition to committed and differentiated cells, a subpopulation with stem cell characteristics. AF-derived stem cells (AFS) have functions found in mesenchymal stem cells, but in addition, exhibit a potent expansion capacity and plasticity. AFS are able to undergo multi-lineage differentiation and produce progeny indicative of all three germ layers. AREAS COVERED: The experimental approaches available to isolate AFS and their potential for tissue engineering, the repair of organs through cell replacement and tissue regeneration. EXPERT OPINION: The deployment of AFS for tissue regeneration offers advantages over the use of embryonic or adult stem cells: i) AF represents a convenient and non-contested source for obtaining stem cells; ii) their derivation is relatively simple and rapid; iii) no feeder layers are required for their cultivation; iv) they display no spontaneous differentiation in culture; and v) their stem cell phenotype is not affected by long-term storage. The application of AFS for tissue replacement therapies in vivo is at a very early stage, but existing studies indicate great potential for clinical use.     

Autocorrelation length of normal and myopathic human myocardium measured by acoustic microscopy: implications for clinical ultrasonic tissue characterization

The pattern of acoustic speckle present in ultrasonic images of tissue may be affected by pathological changes in the tissue. To investigate the possibility for such an effect in dilated cardiomyopathy, we examined acoustic micrographs of frozen sections of normal (autopsy) and myopathic (transplant recipient) human myocardium. The resulting images were digitized, and the major axis autocorrelation length calculated for each image. For the normal specimens (n = 75, four patients), the mean autocorrelation length was 37 +/- 18 microns, with the maximum value of 92 microns. The myopathic specimens (n = 64, four patients) had a mean of 52 +/- 19 microns, with a maximum of 100 microns. We conclude that the changes in structure between normal and myopathic myocardium are significant (p less than 0.001), but on so small a scale in relation to the wave length of clinical ultrasound that there will only be changes in the intensity of the backscattered signal, not the pattern of speckle.     

Influence of histochemical preparation on acoustic parameters of liver tissue: A 5-MHz study

In this study the influence of various histological techniques on the acoustic parameters of liver tissue was investigated. Radiofrequency (RF) echographic data were obtained in vitro from 21 liver samples taken from 8 white New Zealander rabbits. The samples were measured in four different subsequent histological tissue processing conditions (freshly excised, 4% buffered formalin fixed, after it went through a paraffin cycle and after staining with hematoxylin and eosin). The acoustic parameters that were obtained from the rf data were velocity of sound, slope of the attenuation coefficient versus frequency between 1.9 and 6.9 MHz, attenuation coefficient at 4.4 MHz, slope of the backscattering spectrum between 1.9 and 6.9 MHz, and intercept of the backscattering spectrum. It was found that fixation by formalin preserves the acoustic properties of the tissue to a reasonable extent. Embedding in paraffin and deparaffinizing induces large changes in the acoustic properties of the tissue. As an alternative, freezing prior to cutting, rather than the paraffin cycle, was investigated also in 10 liver samples obtained from 4 New Zealander rabbits. This method produced no significant changes of the acoustic parameters and should therefore be preferred in acoustic microscopy.     

The sensitivity of thermography to temperature changes in breast tissue

This study evaluates the ability of thermography to i) detect temperature changes in a lactating breast in response to the application of therapeutic ultrasound; ii) identify an area of raised temperature in a breast affected by mastitis; and iii) detect temperature changes in the affected area in response to the application of ultrasound. An area of raised temperature (36 degrees C), corresponding with the area of inflammation, was identified in a mastitic breast. Eighty-seven per cent of this area had returned to normal resting breast temperature (33 degrees C) within 30 minutes of application of pulsed ultrasound. The use of thermography to evaluate temperature changes in mastitic breast tissue before and after the application of ultrasound is supported.     

Intratracheal administration of fentanyl: Pharmacokinetics and local tissue effects

Objective To study the pharmacokinetics and local tissue effects resulting from the intratracheal administration of preservative-free fentanyl.Design Prospective, randomized, blinded and controlled animal study.Setting University research laboratory.Subjects Eighteen adult male New Zealand rabbits.Interventions Preservative-free fentanyl citrate or normal saline was administered by the intratracheal (i.t.) and intravenous (i.v.) routes to randomized groups of rabbits. The animals were killed at 24, 48 and 72 h following administration.Measurements and main results Plasma concentrations of fentanyl were measured before administration and at 2, 5, 10, 30, 60 and 120 min following administration by a specific radioimmunoassay. A detailed histological examination of the lung and tracheal tissue was performed to identify local side effects. There were no significant differences in the plasma fentanyl concentrations resulting from the i.v. or i.t. route of administration. In both groups, the concentrations of fentanyl were within the therapeutic range (i.t. 2.37 ng/ml, i.v. 2.53 ng/ml) by 2 min after injection and reached a maximum concentration within 5 min. The bioavailability of i.t. fentanyl was 71%. Microscopic examination of the respiratory system did not show significant differences between the two random groups overall. However, in the sub-group of animals killed at 24 h, more animals in the i.t. group showed signs of inflammation in the lung parenchyma.Conclusions There is rapid absorption of fentanyl following i.t. administration. Pharmacokinetic parameters for fentanyl were not significantly altered by the route of administration. Although there were no signs that i.t. administration of preservative-free fentanyl produces lung injury, a transient and mild inflammatory response was detected at 24 h after administration     

Improvement of postoperative hypocalcemia by repeated allotransplantation of parathyroid tissue without anti-rejection therapy.

Long lasting postoperative hypocalcemia, an uncomfortable complication of a thyroid operation for hyperthyroidism, was treated with allotransplantation of parathyroid tissue. Small pieces of the parathyroid tissue offered from two unrelated donors were transplanted to an 18-year-old male with severe postoperative hypoparathyroidism. Prednisolone was given for 2 days, but no other immunosuppressive drugs were used. The remaining tissue was stored in frozen for the repeat transplantation. The functional activity of the frozen tissue was determined by the production of parathyroid hormone in the tissue culture medium adjusted to appropriate concentration of calcium. Loss of the graft function, probably due to rejection, was supplemented with repeated grafting. Hypocalcemia was improved by three times of transplantation using frozen tissue (once) and fresh tissue (twice). This preliminary trial demonstrates that the tissue transplantation of the parathyroid gland is effective to lessen the symptoms and medication of postoperative hypoparathyroidism.     

同种肾组织移植改善肾性贫血的基因水平检测

背景：促红细胞生成素分泌不足使慢性肾衰所引起的贫血（肾性贫血）难以改善。目的：观察肾组织移植对肾性贫血大鼠促红细胞生成素基因表达的影响。方法：80只Wistar大白鼠随机均正常对照组,病例对照组、重组人类促红细胞生成素组和肾组织移植组,后3组建立慢性肾功能衰竭动物模型。结果与结论：60d时移植组血红蛋白水平及血清促红细胞生成素高于病例对照组（P〈0.05）,与重组人类促红细胞生成素组比较,差异无显著性意义（P〉0.05）。移植组肾组织EPO mRNA表达显著高于病例对照组,差异有显著性意义（P〈0.05）。提示肾组织移植改善肾性贫血的作用机制是促进肾组织EPO mRNA的基因表达,使肾脏合成促红细胞生成素增多,从而提高血红蛋白水平。     

Measuring quality of life with the sickness impact profile: A pilot study

A pilot-study was done to investigate the applicability of the sickness impact profile (SIP) in ex-ICU patients. For this study 221 consecutively admitted patients were reviewed retrospectively after excluding children, deceased pateints and readmissions. SIP was assessed in these patients by either interview or questionnaire. These were divided into three groups: i) Patients interviewed at home (n=26). ii) Patients receiving the SIP-questionnaire by mail (n=93). iii) As for group ii, but after receiving a telephone invitation to participate (n=102). Highest mean SIP-score was found in group i (16.3). Groups ii and iii scored 10.2 and 7.9 respectively. Analysis of variance demonstrated overall SIP-scores of these groups to be significantly different. The response in group iii (77%) was significantly higher compared to group ii (56%). Data collection in Group i appeared to be most expensive costing $13.20 per patient, followed by group iii ($3.79) and group ii ($2.56). It is concluded that the self-administered SIP is suitable for measuring outcome in ICU-patients and is much cheaper than the direct interview technique. The 3 different approaches should be considered as independent methods of which individual results cannot be compared. The response can be improved significantly by calling the patients before sending the questionnaire.     

Electrodiagnosis of mild carpal tunnel syndrome

Electrophysiologic tests have been reported to detect mild carpal tunnel syndrome (CTS). Such tests include (i) absolute palmar latency of median wrist segment; (ii) comparison of median and radial distal sensory latencies in digit I; (iii) comparison of median and ulnar distal sensory latencies in digit IV; (iv) comparison of median and ulnar palmar latencies; (v) comparison of median and ulnar sensory potential amplitudes in digits II and V. To clarify the clinical utility of these tests, the parameters of all five tests were determined across four carefully established patient subgroups: group A, controls; group B, CTS referrals with normal nerve conduction studies (NCS) and normal needle electromyography (EMG); group C, CTS referrals with abnormal NCS and normal EMG; group D, CTS referrals with abnormal NCS and abnormal EMG. Special attention was focused on patients in group B who represent the diagnostic dilemma. In group B, tests ii and iii each yielded abnormal results in 44% of hands, while the combination of tests ii and iii yielded abnormal results in 51% of hands.     

The potential of amniotic fluid stem cells for cellular therapy and tissue engineering

Introduction: Foetal cells present in amniotic fluid (AF) have been used for many years to perform prenatal genetic screening. Recent reports suggested that these cells might have additional benefits. AF contains, in addition to committed and differentiated cells, a subpopulation with stem cell characteristics. AF-derived stem cells (AFS) have functions found in mesenchymal stem cells, but in addition, exhibit a potent expansion capacity and plasticity. AFS are able to undergo multi-lineage differentiation and produce progeny indicative of all three germ layers.

Areas covered: The experimental approaches available to isolate AFS and their potential for tissue engineering, the repair of organs through cell replacement and tissue regeneration.

Expert opinion: The deployment of AFS for tissue regeneration offers advantages over the use of embryonic or adult stem cells: i) AF represents a convenient and non-contested source for obtaining stem cells; ii) their derivation is relatively simple and rapid; iii) no feeder layers are required for their cultivation; iv) they display no spontaneous differentiation in culture; and v) their stem cell phenotype is not affected by long-term storage. The application of AFS for tissue replacement therapies in vivo is at a very early stage, but existing studies indicate great potential for clinical use.