肾上腺素对红细胞血红蛋白携氧能力的影响

目的:探讨肾上腺素是否影响红细胞血红蛋白的携氧能力。方法:应用拉曼光谱扫描光谱分析肾上腺素作用下及酸性环境下的红细胞氧合血红蛋白的改变。采集健康人红细胞,悬浮于不同pH值(pH 5.0、pH7.4)等渗PBS缓冲液,肾上腺素预处理红细胞,在拉曼光谱点扫描分析检测单个活态红细胞内血红蛋白特征光谱变化。结果:酸性环境下,红细胞的氧合血红蛋白特征峰在肾上腺素的作用下显著升高(P0.05);而在正常pH时,肾上腺素的这一作用不明显(P0.05)。结论:肾上腺素在酸性环境时增加红细胞血红蛋白的氧合力,减少氧从血红蛋白的解离,不利于缺氧组织供氧。     

激光共焦拉曼光谱法对不同pH环境下鳄鱼红细胞内血红蛋白结构功能的研究

目的：使用激光共焦拉曼光谱扫描术对不同pH环境下活态鳄鱼红细胞内血红蛋白结构功能的变化进行测定研究。以期揭示鳄鱼红细胞及其血红蛋白随不pH环境下的结构功能状态的变化规律。方法：鳄鱼血活体采集后用肝素抗凝,无菌分离出细胞后加入pH值分别为2.55、3.02、4.39、5.50、6.58、7.13、7.82、8.31、8.80、10.0的细胞缓冲溶液中,静置20 min后,使用拉曼光谱仪进行扫描获得红细胞拉曼谱,通过分析测定获得其血红蛋白的分子结构与携氧功能随pH值变化的情况。结果：1.鳄鱼红细胞血红蛋白在pH=3.02~pH=8.80均能保持正常的生理活性;2.鳄鱼红细胞中氧合蛋白占总蛋白百分比在pH=7.13和pH=7.80条件下最高,随着pH的升高或降低,红细胞中氧合蛋白占总血红蛋白的总量百分比均下降;3.鳄鱼红细胞通过调节携氧血红蛋白比例来适应不同pH值条件下的携氧需求,在正常pH条件下无需所有血红蛋白即可完成携氧功能,而在酸性条件下可以使用全部血红蛋白完成携氧功能;4.鳄鱼红细胞抗酸性能力比抗碱性能力强。结论：研究表明鳄鱼红细胞的血红蛋白具有比人类红细胞更强的环境适应能力和更强的携氧能力,对人造血的研究有指导作用。     

局部振动刺激对肱二头肌血红蛋白的影响

目的探讨局部振动刺激过程对肱二头总血红蛋白和氧合血红蛋白的影响。方法利用手臂振动按摩带对12名女性志愿者进行局部振动刺激,用非侵入性近红外光谱仪于振动开始前10 min、振动中10 min与振动停止后15 min采集肱二头肌之总血红蛋白和氧合血红蛋白的数值,并了解每分钟变化趋势;以单因素方差分析进行统计检验。结果振动停止后15 min会使手臂肱二头总血红蛋白显著性增加(P<0.05),振动中10 min与振动停止后15 min均会使手臂肱二头氧合血红蛋白显著性增加(P<0.05);在变化趋势方面,总血红蛋白和氧合血红蛋白约在振动中10 min的第5 min出现最大峰值,且在振动停止后15 min仍持续高于振动刺激前,并呈现稳定状态。结论局部振动刺激会实时提升肱二头总血红蛋白和氧合血红蛋白;此外,必须至少持续振动刺激5 min以上总血红蛋白和氧合血红蛋白数量才会达到最大峰值。     

维生素D受体基因内切酶位点多态与孕妇骨密度和补钙的关系

目的:研究维生素D受体基因内切酶位点多态在孕妇(南京地区)人群中的分布及其对补钙的影响。方法:采用聚合酶链反应(两对引物)扩增维生素D受体基因,再用内切酶BsmI、ApaI、TaqI消化扩增产物,然后琼脂糖凝胶电泳,紫外灯下观察分析和照相,根据带型进行VDR基因分型。骨密度测量采用单光子吸收法。结果:孕妇中等位基因B所占的比例为4.8%(48/1000),b所占的比例为95.2%(952/1000),A所占的比例为22.2%(222/1000),a所占的比例为77.8%(778/1000),T所占的比例为90.3%(903/1000),t所占的比例为9.7%(97/1000);基因型为BB:2.8%(14/500),Bb:4%(20/500),bb:93.2%(466/500),AA:4%(20/500),Aa:36.4%(182/500),aa:59.6%(298/500);TT:82.2%(411/500),Tt:16.2(81/500),tt:1.6%(8/500),符合遗传平衡定律(Hardy-Weinberglaw)即(p q)2=1。BBAAtt=1.6%(8/500),BbAATt=2.4%(12/500),BBAaTt=1.2%(6/500),BbAaTt1.6%(8/500),bbAaTt=11%(55/500),bbAaTT=22.6%(113/500),bbaaTT=59.6%(298/500)。结论:在中国妇女中基因型BBAAtt的人具有较高的骨密度,而bbaaTT基因型的人骨密度较低,但占人群的50%以上,所以中国妇女的平均骨密度较低,大部分孕妇在孕期需要补钙,建议每天摄入600mg～1200mg。     

显微激光共焦拉曼散射仪性能快速测试方法

目的:研究显微激光共焦拉曼散射装置性能快速测定方法,为进行其他样品拉曼测定提供质量保证。方法:利用HoribaJY公司的LabRamINV倒置显微激光共焦拉曼散射仪进行硅片标准样的实验。利用硅片标准样进行系统校正,并利用标准样进行系统性能参数的测定。结果:实验得到硅片标准样位于520.7cm-1的谱线强度随照射激光功率、共焦孔径和时间的变化曲线。结论:520.7cm-1谱线强度随照射激光功率成线性关系,利用该谱线强度的变化可以间接反映照射到样品处的激光功率;520.7cm-1谱线强度随共焦孔径的变化曲线可用于确定不同实验目的共焦孔径尺寸的设置;520.7cm-1谱线强度随时间的变化曲线反映系统的稳定性,从而判断是否适合进行其他样品的测试。     

褐藻糖胶对人乳腺癌MCF-7细胞的增殖抑制和凋亡诱导作用

目的：研究褐藻糖胶对体外培养的人乳腺癌MCF-7细胞增殖及凋亡的影响，并探讨其凋亡机制。方法:不同浓度(100 mg/L、300 mg/L、500 mg/L、1 000 mg/L)褐藻糖胶处理MCF-7细胞48h后，应用四甲基偶氮唑蓝（MTT）法检测细胞的增殖；Hoechst 33258染色、琼脂糖凝胶电泳法观察细胞的凋亡的形态学及生化改变；逆转录聚合酶链反应（RT-PCR）和蛋白印迹分别测定细胞〖STBX〗bcl-2〖STBZ〗和bax mRNA及其蛋白的表达。结果:不同实验浓度(100 mg/L、300 mg/L、500 mg/L、1 000 mg/L)的褐藻糖胶均能抑制MCF-7细胞的增殖（P<0.01），抑制率呈浓度依赖性增大；褐藻糖胶诱导MCF-7细胞凋亡数增加，且可见明显的、凋亡特有的DNA梯形条带；在褐藻糖胶存在下，〖STBX〗 bcl-2〖STBZ〗 mRNA和蛋白表达减少，而bax mRNA和蛋白表达增加，Bcl-2/Bax比值下降（P<0.05）。结论:褐藻糖胶可抑制MCF-7细胞增殖，且诱导其凋亡，其凋亡机制是下调Bcl-2和上调Bax蛋白的表达。     

数字式无线肌血氧监测仪的研制

目的研制微型肌血氧检测系统，可实时地对人体肌肉组织的血氧含量变化进行无损检测。方法利用与机体氧合血红蛋白和还原血红蛋白对光吸收谱不同，采用数字信号处理技术，系统硬件采用自行设计的反射式传感器、数字信号处理模块和高速率、低功耗小型无线数传模块，设计数字式无线肌血氧监测仪。参数调整全部软件化，数据计算模块化。利用血模型实验和初步人体试验检验该仪器。结果通过血模型实验，在模型中氧合血红蛋白含量不变时，改变血容积，在血通道上出现明显变化；在模型中血容积不变时，通过改变溶液中的氧含量，在氧通道上出现明显变化，且在耗氧结束时能恢复到原始水平。人体试验，静息和运动时，血、氧含量发生变化；停止运动后，肌肉组织耗氧减少，受试者血、氧含量均逐渐恢复到静息水平。结论该仪器具有灵敏度高、性能稳定、抗干扰性好等特点。该设计方案可行，有望用于对运动员有氧代谢功能和运动强度的评定。     

308nm准分子激光对人角质形成细胞活力及bFGF表达与分泌的影响

目的:探讨308 nm准分子激光对人角质形成细胞(hKCs)活力及碱性成纤维细胞生长因子(bF-GF)表达与分泌的影响。方法:体外培养人角质形成细胞,免疫荧光法鉴定细胞。分别予不同剂量308 nm准分子激光及311 nm窄谱中波紫外线(NB-UVB)照射,MTT法检测hKCs细胞活力,RT-PCR法和ELISA法分别检测bFGF表达和分泌水平,并与NB-UVB进行对比。结果:细胞鉴定证实为角质形成细胞。308 nm准分子激光剂量达600 mj/cm2,NB-UVB剂量达500 mj/cm2时hKCs活力下降;2种光源在一定剂量范围内照射均可使hKCs表达、分泌bFGF水平增加,并呈剂量依赖性;308 nm准分子激光诱导hKCs分泌bFGF的水平高于NB-UVB。结论:308 nm准分子激光照射对hKCs活力的影响小于NB-UVB;308 nm准分子激光可以通过剂量依赖的方式诱导hKCs表达与分泌bFGF来促使白癜风复色,与NB-UVB相比效果更佳。     

雌激素对人乳腺癌细胞株MCF-7 KLK 6 mRNA和蛋白表达水平的影响

 目的： 探讨雌激素与其拮抗剂三苯氧胺对雌激素受体阳性人乳腺癌细胞株MCF-7组织激肽释放酶6（KLK 6）mRNA和蛋白（hK6）表达水平的影响。方法: 取生长良好的MCF-7细胞，在不同浓度的雌激素（17-βE2）和三苯氧胺存在下培养72 h后，收集细胞，分别采用荧光定量逆转录-聚合酶链反应和流式细胞术，检测KLK6 mRNA和蛋白表达水平的变化。结果: 实时荧光定量RT-PCR表明，当17-βE2浓度在10-12 mol/L时，与乙醇对照组无明显差异(P>0.05), 17-βE2浓度在10^-10 mol/L和10-8 mol/L时，KLK6 mRNA的相对含量显著少于乙醇对照组；流式细胞术分析表明, 代表hK6含量的相对荧光强度（AFI）显著低于乙醇对照组（P<0.01）。三苯氧胺（10^-6 mol/L）作用组KLK6 mRNA 的相对含量显著高于对照组（P<0.01）；hK6含量与对照组相比差异显著（P<0.01）。结论: 雌激素对MCF-7 KLK6 mRNA和蛋白表达水平有一定的下调作用，而三苯氧胺作用相反。     

氧化低密度脂蛋白促进血管内皮细胞表达基质金属蛋白酶-2

目的:用氧化低密度脂蛋白干预培养的人脐静脉内皮细胞, 观察内皮细胞表达基质金属蛋白酶-2(MMP-2)的改变。方法:用胶原酶消化法培养人脐静脉内皮细胞, 分组后分别予以25mg/LLDL、25mg/LOX-LDL进行干预。提取细胞总RNA和总蛋白分别进行RT-PCR、Westernblotting检测, 同时收集条件培养基行酶谱法(zymography)检测MMP-2的活性。结果:经mRNA、蛋白和酶活性检测, 发现25mg/LOX-LDL干预时, MMP-2的表达水平明显高于对照和25mg/LLDL。结论:OX-LDL能促进内皮细胞表达MMP-2, 这提示血管细胞外基质的降解在氧化脂蛋白诱发的动脉粥样硬化斑块产生和破裂机制中起着一定作用。     

Study of forensic and clinical source hemoglobin interference with the duPont aca ethanol method

The enzymatic alcohol (ethanol, ALC) analytical pack for the duPont aca is intended primarily for the analysis of serum or plasma samples containing no visibly detectable free hemoglobin (non-hemolyzed samples). It has become apparent, through clinical and medicolegal consultation, that this methodology has been applied for assay of ethanol content in hemolyzed samples (sample sources: clinical and forensic cases). The influence of sample free hemoglobin on the ace ethanol method was investigated. With 'hemolyzed' clinical samples (containing primarily oxyhemoglobin) a significant decrease was found in known ethanol concentration at 0.5 g per dl and greater of hemoglobin with the influence being greatest at lower ethanol concentration. With forensic samples (containing varying amounts of oxy-, reduced, and methemoglobins), the same observation was made with measured decrease in ethanol concentration beginning at 0.1 g per dL hemoglobin and greater. In essence, the higher the sample hemoglobin concentration the greater would be the effect in decreasing the apparent ethanol concentration. Preparation of protein-free trichloroacetic acid supernatants and their assay on the aca and by head-space gas chromatography resulted in excellent recovery of the sample ethanol. It is concluded that the effect of sample free hemoglobin is spectral in nature, and its presence results in potential and real overblanking for the aca ethanol enzymatic reaction sequence.     

Hemoglobin determination in plasma or serum by first-derivative recording spectrophotometry. Evaluation of the procedure of Soloni, Cunningham, and Amazon

The authors tested the plasma hemoglobin measurement procedure of Soloni, Cunningham, and Amazon (Am J Clin Pathol 1986;85:342-347), which uses recording derivative spectrophotometry. By this technique, the authors were able to measure plasma or serum hemoglobin down to a level of 10 mg/L (1 mg/dL). The method was found to be quantitative and not affected by bilirubin or lipemia. Methemoglobin did not interfere with the assay. The observed first derivative maxima and minima were reproducible but instrument dependent. The authors applied this technique to the measurement of serum hemoglobin in 100 healthy human subjects. They observed a geometric mean value of 68 mg/L (6.8 mg/dL) with the actual range 21-189 mg/L (2.1-18.9 mg/dL). The authors confirm that this technique is a simple, rapid, and reproducible method for determining plasma or serum hemoglobin.     

Treatment of homozygous sickle cell disease with pentoxifylline

This case report describes the sustained symptomatic and hematologic improvement in a 21-year-old woman with homozygous sickle cell (ss) disease during treatment with pentoxifylline, 400 mg three times daily after meals. Pain crises decreased from six to zero per year, hemoglobin level rose from 8.4 g/dL to 11.4 g/dL, hematocrit rose from 24.8% to 34.8%, lactate dehydrogenase level decreased from 375 IU/L to 322 IU/L, and total bilirubin level decreased from 1.8 mg/dL to 1.6 mg/dL. Mean corpuscular hemoglobin increased from 21.6 pg to 30 pg and mean corpuscular hemoglobin concentration increased from 24.1 g/dL to 34.5 g/dL. These changes were sustained for seven years except for a brief self-imposed hiatus in therapy during which period a pain crisis occurred. Further increase in pentoxifylline dosage to 400 mg four times daily did not result in any further improvement in these hematologic parameters. These results suggest that pentoxifylline reduces hemolysis in SS patients with a resulting improvement in anemia and a reduction in or elimination of pain crises.     

Hematology of the Mediterranean population of sea turtle (Caretta caretta): comparison of blood values in wild and captive, juvenile and adult animals

In order to establish baseline hematological and biochemical values in loggerhead turtles from the Mediterranean Sea, 84 specimens were sampled, comprising 24 wild turtles in good health at the time of capture and 60 turtles tested after indoor rehabilitation at the Sea Turtle Rescue and Rehabilitation Centre of the Zoological Station Anton Dohrn in Naples, Italy. The following parameters were evaluated: red cell counts (RBC, 488–575?×?10³/μL), white cell counts (WBC, 17–24?×?10³/μL) and thrombocyte counts (TBC, 19–49?×?10²/μL), hemoglobin (Hb, 8–14?g/dL), hematocrit (Ht, 23–34%), mean corpuscular volume (MCV, 487–723?fL), mean corpuscular hemoglobin (MCH, 170–261?pg), mean corpuscular hemoglobin concentration (MCHC, 34–42%), white and red blood cell differential counts, and a panel of hematochemical tests, composed of glucose (97–164?mg/dL), cholesterol (74–144?mg/dL), blood urea nitrogen (35–200?mg/dL), uric acid (1–2.7?mg/dL), total bilirubin (0.20–0.40?mg/dL), GOT (44–184?IU/L) and GPT (6?IU/L) transaminases, calcium (6.7–8.7?mg/dL), and magnesium (3.6–5.4?mEq/L). Comparisons of the statistically analyzed data from the turtles which were divided into groups on the basis of age and/or lifestyle (wild or captive) revealed that erythroid parameters attained higher values in captive turtles. This suggested a positive influence of the rich and complete diet fed in captivity upon the hemopoietic process of the turtle. On the other hand, data suggest a more intense and active hemopoiesis in young turtles, compared to adult specimens.     

Diffuse reflectance spectra of the palpebral conjunctiva and its utility as a noninvasive indicator of total hemoglobin

The palpebral conjunctiva is an attractive location to qualitatively examine for the presence of anemia; however, this method of diagnosis has not been shown to be accurate. A spectroscopic examination of the palpebral conjunctiva enables the use of a quantitative parameter as a basis for diagnoses. Visible range diffuse reflectance spectra from the palpebral conjunctiva are examined from 30 patients and hemoglobin levels are extracted from these signatures using both a partial least-squares (PLS) multivariate regression model and a discrete spectral region model. Hemoglobin concentration derived from both these models is compared to an in vitro measurement of hemoglobin. Root mean squared errors of cross validation for the two analytical methods are 0.67 g/dL and 1.07 g/dL, respectively. Conjunctival reflectance spectra coupled with a PLS analysis achieve an enhanced specificity and sensitivity for anemia diagnoses over reported observational studies using the palpebral conjunctiva and achieve improved accuracy to other reported methods of noninvasive hemoglobin measurement.     

Data preprocessing and partial least squares regression analysis for reagentless determination of hemoglobin concentrations using conventional and total transmission spectroscopy

Visible-near infrared spectroscopy was successfully used for the determination of total hemoglobin concentration in whole blood. Absorption spectra of whole blood samples, whose hemoglobin concentrations ranged between 6.6 and 17.2 g/dL, were measured from 500 to 800 nm. Two different types of transmission were measured: conventional transmission spectroscopy which collected primarily collimated radiation transmitted through the sample, and total transmission spectroscopy which used an integrating sphere to collect all scattered light as well. Different preprocessing techniques in conjunction with a partial least squares regression calibration model to predict hemoglobin concentrations were applied to the above two types of transmission. Depending on different preprocessing methods, the standard error of predictions ranged from 0.37 to 2.67 g/dL. Mean centering gave the most accurate prediction in our particular data set. Preprocessing methods designed for compensation of the scattering effect produced the worst results contrary to expectations. For univariate analysis, better prediction was achieved by total transmission measurement than by conventional transmission measurement. No significant difference was observed for multivariate analysis on the other hand. Careful selection of the data preprocessing methods and of the multivariate statistical model is required for reagentless determination of hemoglobin concentration in whole blood.     

Effect of hemoglobin concentration variation on the accuracy and precision of glucose analysis using tissue modulated, noninvasive, in vivo Raman spectroscopy of human blood: a small clinical study

Tissue modulated Raman spectroscopy was used noninvasively to measure blood glucose concentration in people with type I and type II diabetes with HemoCue fingerstick measurements being used as reference. Including all of the 49 measurements, a Clarke error grid analysis of the noninvasive measurements showed that 72% were A range, i.e., clinically accurate, 20% were B range, i.e., clinically benign, with the remaining 8% of measurements being essentially erroneous, i.e., C, D, or E range. Rejection of 11 outliers gave a correlation coefficient of 0.80, a standard deviation of 22 mg/dL with p<0.0001 for N=38 and places all but one of the measurements in the A and B ranges. The distribution of deviations of the noninvasive glucose measurements from the fingerstick glucose measurements is consistent with the suggestion that there are at least two systematic components in addition to the random noise associated with shot noise, charge coupled device spiking, and human factors. One component is consistent with the known variation of fingerstick glucose concentration measurements from laboratory reference measurements made using plasma or whole blood. A weak but significant correlation between the deviations of noninvasive measurements from fingerstick glucose measurements and the test subject's hemoglobin concentration was also observed.     

Six-month hemoglobin concentration and its association with subsequent mortality among adults on antiretroviral therapy in lusaka, zambia

ABSTRACT:: Little is known about changes in hemoglobin concentration early in the course of antiretroviral therapy and its subsequent relation to survival. We analyzed data for 40,410 HIV-infected adults on antiretroviral therapy in Lusaka, Zambia. Our main exposure of interest was 6-month hemoglobin, but we stratified our analysis by baseline hemoglobin to allow for potential effect modification. Patients with a 6-month hemoglobin <8.5 g/dL, regardless of baseline, had the highest hazard for death after 6 months (hazard ratio: 4.5; 95% confidence interval: 3.3 to 6.3). Future work should look to identify causes of anemia in settings such as ours and evaluate strategies for more timely diagnosis and treatment.     

Anemia in HIV-infected patients receiving highly active antiretroviral therapy.

BACKGROUND: Anemia is common in HIV infection, particularly in advanced disease states. We wished to determine how highly active antiretroviral therapy (HAART) and other factors affected the level of hemoglobin in HIV infection. METHODS: We analyzed data from 905 patients receiving care at Johns Hopkins in Baltimore, Maryland after July 1, 1996. Analyses were done of hemoglobin levels obtained at baseline and during 1 year of follow-up in patients who received and did not receive a HAART regimen. Use of HAART and other demographic and clinical factors were examined. RESULTS: Eleven percent of patients had a hemoglobin count <10 g/dL, 27% had a hemoglobin count 10 to 12 g/dL, and 21% had a hemoglobin count of >14 g/dL at baseline before HAART was started. During 1 year of follow-up, use of HAART was associated with a hemoglobin levels >14 g/dL in 42% of patients, irrespective of use of zidovudine as part of HAART regimen, compared with 31% of patients who did not use HAART. In multivariate analysis, use of HAART was strongly associated with not having anemia during 1 year of follow-up, adjusting for patient gender, race, injection drug use history, baseline CD4 and HIV-1 RNA levels, and anemia treatments. CONCLUSIONS: HAART is an effective treatment of the anemia of HIV infection. Patients who continue to have symptomatic anemia while receiving HAART may need additional intervention.     

Uncertainty in estimating blood ethanol concentrations by analysis of vitreous humour

AIMS: To determine the concentrations of ethanol in femoral venous blood (FVB) and vitreous humour (VH) obtained during forensic necropsies. The ratios of ethanol concentrations in VH and FVB, the reference interval, and the associated confidence limits were calculated to provide information about the uncertainty in estimating FVB ethanol concentrations indirectly from that measured in VH. METHODS: Ethanol concentrations were determined in specimens of FVB and VH obtained from 706 forensic necropsies. The specimens were analysed in duplicate by headspace gas chromatography (HS-GC), with a precision (coefficient of variation) of 1.5% at a mean ethanol concentration of 500 mg/litre. The limit of detection of ethanol in body fluids by HS-GC in routine casework was 100 mg/litre. RESULTS: In 34 instances, ethanol was present in VH at a mean concentration of 154 mg/litre, whereas the FVB ethanol concentration was reported as negative (< 100 mg/litre). These cases were excluded from the statistical analysis. The concentration of ethanol in FVB was higher than in VH in 93 instances, with a mean difference of 160 mg/litre (range 0 to 900). The mean concentration of ethanol in FVB (n = 672) was 1340 mg/litre (SD, 990) compared with 1580 mg/litre (SD, 1190) in VH. The arithmetic mean VH/FVB ratio of ethanol was 1.19 (SD, 0.285) and the 95% range was 0.63 to 1.75. The mean and SD of the differences (log VH - log FVB) was 0.063 (SD, 0.109), which gives 95% limits of agreement (LOA) from -0.149 to 0.276. Transforming back to the original scale of measurement gives a geometric mean VH/FVB ratio of 1.16 and 95% LOA from 0.71 to 1.89. These parametric estimates are in good agreement, with a median VH/FVB ratio of 1.18 and 2.5th and 97.5th centiles of 0.63 and 1.92. CONCLUSIONS: The ethanol distribution ratios (VH/FVB) show wide variation and this calls for caution when results of analysing VH at necropsy are used to estimate the concentration in FVB. Dividing the ethanol concentration in VH by 2.0 would provide a very conservative estimate of the ethanol content in FVB, being less than the true value, with a high degree of confidence.