低强度He-Ne激光对红细胞胞浆内游离钙离子浓度的影响

低强度He-Ne激光对红细胞变形性的影响已得到广泛的认可和应用,但其具体调节机制尚不明确,通过研究低强度He-Ne激光对红细胞胞浆内钙离子浓度的影响,探讨其对红细胞变形性影响的机制。采用A23187处理过的红细胞,分别用5 mW和9 mW激光照射后,观察红细胞胞浆内钙离子浓度变化。结果显示:低强度He-Ne激光照射后的红细胞与无照射组的红细胞相比,红细胞胞浆内钙离子浓度显著降低,但在本实验中钙离子浓度的降低与激光照射剂量无显著相关。由此得出结论:降低红细胞胞浆内钙离子浓度可能是低强度He-Ne激光调节红细胞变形性的重要机制。     

小鼠网织红细胞微观流变学特性的研究

按Bishop方法,在小鼠血液里诱导生成大量网织红细胞,然后提取网织红细胞,对其电泳率、渗透脆性、膜的流动性、细胞的变形能力和取向性进行了系统研究。研究结果表明网织红细胞在转变为成熟红细胞的短短时间内,其微观流变学特性发生了明显的变化：电泳率变小、渗透脆性变好、膜的流动性变大、细胞的变形能力变强、取向性变好,最终发育成具有全面功能的成熟红细胞。     

低强度氦氖激光对缺血性疾病患者红细胞变形性影响的临床研究

目的：探讨低强度氦氖激光对临床中风等缺血性疾病患者红细胞变形能力影响情况。方法：临床采集不同缺血性疾病患者新鲜血样,检测各组红细胞变形能力情况。对中风患者血样进一步分组,给予不同剂量的氦氖激光处理,讨论临床低强度氦氖激光量效关系,探讨具有临床疗效的有效剂量情况。结果：各疾病病组红细胞变形能力较正常对照组红细胞变形能力为低;缺血疾病组样品经低强度氦氖激光照射后,其红细胞变形能力较未经激光照射组有显著不同。结论：显示体外人红细胞变形能力因患者疾病情况、红细胞状态、低强度氦氖激光临床治疗输出功率等因素有不同。     

低强度He-Ne激光血管内照射影响糖尿病兔血液流变性质的可能因素分析

本实验在前工作的基础上,观察对实验性糖尿病兔进行两次低强度He-Ne激光血管内照射后,血中甘油三酯、总胆固醇、红细胞膜总磷脂的动态变化,分析有关参数与红细胞变形能力之间的关系,以期探讨低强度激光血管内照射影响血液流变性质的可能因素。实验结果表明,经两次激光照射后,1．第1天糖尿病兔的红细胞变形能力即见改善,4天-7天后接近注药前水平。2．与其相应,甘油三酯和胆固醇在激光照射后第4天即回复至正常水平。3．红细胞膜总磷脂激光照射后渐升高,至第7天与注药前无明显差异。文章讨论了有关实验指标影响红细胞变形能力的可能性。     

影响低强度He-Ne激光对红细胞生物效应诸因素分析与有关问题讨论

在分析有关低强度He-Ne激光对与红细胞的作用的临床报道与细胞生物学研究相关文献基础上,研究影响低强度激光临床疗效和红细胞对生物效应的相关因素,讨论机体状态、病变因素、激光剂量强弱等影响因素的实际临床意义,低强度He-Ne激光在临床安全使用等问题亦被讨论。     

低功率He-Ne激光照射下细胞内活性氧自由基的产生和游离Ca^2＋浓度的研究

在临床应用中,低功率He-Ne激光(632.8 nm)能促进骨骼肌修复,加速创伤愈合,降低牙齿的超敏感性,减缓疼痛等.大量研究表明:低功率He-Ne激光能调节细胞的众多行为,如细胞增殖、分泌、迁移、粘附、蛋白质合成和基因表达等.但低功率He-Ne激光调节细胞行为的分子机制并未阐明,考察低功率He-Ne激光照射后细胞内活性氧自由基的产生水平和游离ca2 浓度是否会发生变化,通过激光扫描共聚焦显微镜,分别利用H:DCFDA和Fluo-3/AM这两种荧光探针,检测到经He-Ne激光照射后,肺腺癌细胞内活性氧自由基的水平上调以及游离Ca2 浓度增加.该研究为低功率He-Ne激光的生物光刺激效应提供了可能的分子机理.     

低强度激光照射对老龄小鼠的抗氧化系统的影响

目的：研究低强度激光照射对机体抗氧化系统的影响。方法：采用不同剂量的低强度He-Ne激光照射老龄小鼠肝区,照射后观察T－SOD,Cn-SOD,Mn-SOD,T-AO,GSH-PX,GSH,MDA等指标。结果：能量密度分别为2．48J／cm^2,12．48J／cm^2,18．72J／cm^2的低强度He-Ne激光可提高,Cu,Zn-SOD,Mn-SOD,总SOD,及GSH－PX活性,可提高GSH含量和总抗氧化能力,降低MDA含量,其中以能量密度为18．72J／cm^2最为显著,使老龄小鼠以上各项指标恢复接近正常水平。结论：低强度激光照射可以提高机体抗氧化酶的活性和抗氧化剂的含量,增强机体抗氧化能力。     

He-Ne激光照射对血液及其组分荧光光谱影响的实验研究

为研究弱激光照射对人血液携氧能力的影响及机制,我们用荧光仪分别测量了He-Ne激光照射前后正常血液及其组分（血浆、红细胞）的荧光光谱,研究了激光照射导致的光谱变化,并分析了光谱变化与血液携氧能力改变的关系。实验结果显示：全血液标本在490nm及614nm附近有荧光峰值;血浆的荧光则主要分布在420－500nm之间;红细胞在500nm及614nm附近有荧光。He-Ne激光照射后,全血液及红细胞在614nm处的荧光谱都有较明显的变化,且较相似。由此可得出结论,He-Ne激光照射可影响血液的携氧能力。     

低功率He-Ne激光照射黑曲霉的方法与效应

目的:探索低功率He-Ne激光对柠檬酸生产菌黑曲霉诱变育种的简易方法。方法:应用带扩束镜的低功率He-Ne激光装置,在无菌条件下对柠檬酸生产菌黑曲霉的单孢子膜进行不同时间的垂直照射,无菌水洗脱,指示性平板筛选,液体培养,测定黑曲霉诱变菌株的柠檬酸产量。结果:不同时间的激光照射黑曲霉单孢子,其存活率与激光照射时间没有线性关系。激光照射6min,黑曲霉M2代产酸的正变率高达37.5%,而此时的存活率也高达40.0%。获得了黑曲霉柠檬酸产酸率提高了10%的突变菌株,为黑曲霉的遗传育种提供了材料。结论:这种方法便于在无菌条件下操作,保证了激光照射黑曲霉单孢子的均匀性,是一种简便易行的微生物诱变育种方法。     

不同发育阶段红系造血细胞的生物力学特性和血液流变特性的变化

用可诱发小鼠贫血的病毒(anemia-inducing strain friend’s virus, FVA)感染BALB/c小鼠, 15 d后取其脾脏, 分离出原红细胞, 在加促红细胞生成素(EPO)等的培养基中培养12, 24, 48 h, 获得大量相对同步化的早幼红细胞、中幼红细胞、晚幼红细胞, 研究了不同发育阶段的上述早期红系造血细胞的生物力学及血液流变学特性的变化规律. 发现不同发育阶段的早期红系造血细胞, 随着发育其电泳率、渗透脆性、膜的流动性和黏弹性都发生了改变, 这种改变与膜脂的组成、膜蛋白、细胞膜的骨架蛋白、脂质分子与蛋白质分子的相互作用有关.     

Alterations of erythrocyte structure and cellular susceptibility in patients with chronic renal failure: effect of haemodialysis and oxidative stress

The aim of this study was to investigate erythrocytes rheological behaviour, membrane dynamics and erythrocytes susceptibility to disintegration upon strong oxidative stress induced by dialysis or by external H(2)O(2) among patients with CRF. EPR spectrometry was used to investigate alterations in physical state of cellular components. Generated ROS production induced: (1) significant increase of membrane fluidity in CRF erythrocytes treated with H(2)O(2) (p<0.005) and at 60 min of haemodialysis (p<0.05), (2) significant decrease of cytoskeletal protein-protein interactions (p<0.005) and (3) cellular osmotic fragility (p<0.0005). H(2)O(2) exacerbated these changes. Erythrocytes from CRF patients have changed rheological behaviour and present higher susceptibility to disintegration. Erythrocytes membrane characteristics indicate that CRF patients possess younger and more flexible cells, which are more susceptible to oxidative stress. This may contribute to the shortened survival of young erythrocytes in CRF patients.     

Comparison of membrane structure,osmotic fragility,and morphology of multiple sclerosis and normal erythrocytes

Erythrocyte membranes from multiple sclerosis (MS) patients and normal individuals were studied by electron spin resonance spectroscopy, osmotic fragility tests, scanning electron microscopy (SEM) and fatty acid analysis of membrane lipids. There was no significant difference in the membrane fluidity between MS and normal erythrocytes using fatty acid spin labels with the nitroxide moiety on carbons 5, 12, or 16 from the carboxyl group. Linoleic acid, which has been reported to decrease the absolute electrophoretic mobility of only MS erythrocytes, increased the fluidity of MS and normal erythrocyte membranes to a similar extent. The osmotic fragility of MS erythrocytes obtained from outpatients was similar to normal control cells but the osmotic fragility of erythrocytes obtained from hospitalized MS patients was greater than normal. Scanning electron microscopy of MS erythrocytes revealed no gross abnormalities. Cells incubated with linoleic acid had transformed from discocytes into sphero-echinocytes with prominent membrane surface indentations but MS and normal erythrocytes appeared identical. Of the fatty acid content of the total lipid extract, erythrocytes from most, but not all, MS hospitalized patients and some patients with other demyelinating diseases had relatively less (P<.001) 18:2 than the normal cells. These results indicate that at least some of the abnormalities reported in MS erythrocytes may only be found in hospitalized patients and may be due to other complications of the disease. They also indicate that the reported abnormal effects of linoleic acid on the electrophoretic mobility of MS erythrocytes may be caused by some other mechanism than an effect on the fluidity of the bilayer.     

Carbamylation of proteins leads to alterations in the membrane structure of erythrocytes

The effect of the sodium cyanate-induced carbamylation (carbamoylation) of proteins in erythrocytes was studied using spin labelling and spectrophotometric methods. The experiments were conducted in whole blood and in erythrocytes in phosphate buffer using 25 mmol/L of sodium cyanate. Lipid membrane fluidity was determined using three spin-labelled fatty acids: 5-, 12- and 16-doxylstearic acids (5-DS, 12-DS, 16-DS). Internal viscosity was measured with Tempamine, using also EPR spectroscopy. Osmotic fragility was determined spectrophotometrically. Incubation of whole blood with sodium cyanate led to an increase in lipid membrane fluidity in the deeper region of the lipid layer, indicated by 12- and 16-doxylstearic acid, and a decrease near the surface (5-DS). Statistically significant results were obtained for the internal viscosity and osmotic fragility of erythrocytes. An increase in internal viscosity and increase in osmotic fragility were found in erythrocytes after incubation of whole blood, as well as in erythrocytes incubated with sodium cyanate in buffer. Alterations in internal viscosity were stronger in erythrocytes incubated with sodium cyanate in blood than in erythrocytes in the buffer. On the other hand, higher osmotic fragility was observed for erythrocytes in the buffer.     

Effects of Proteus mirabilis lipopolysaccharides with different O-polysaccharide structures on the plasma membrane of human erythrocytes

The effects of O33 and O49 P. mirabilis lipopolysaccharides (LPSs) on human erythrocyte membrane properties were examined. Physical parameters of the plasma membrane, such as membrane lipid fluidity, physical state of membrane proteins, and osmotic fragility, were determined. The fluidity of the lipids was estimated using three spin-labeled stearic acids of doxyl derivatives: 5-doxylstearic acid, 12-doxylstearic acid, and 16-doxylstearic acid. All the applied labels locate to different depths of the lipid layer and provide information on the ordering of phospholipid fatty acyl chain mobility. LPSs O49 increased the membrane lipid fluidity in the polar region of the lipid bilayer as indicated by spin-labeled 5-doxylstearic acid. An increase in fluidity was also observed in the deeper region using 12-doxylstearic acid only for O33 LPSs. The highest concentration of O33 LPSs (1 mg/ml) increased the motion of membrane proteins detected by the spin-label residue of iodoacetamide. These results showed different actions of O33 and O49 LPSs on the plasma membrane due to the different chemical structures of O-polysaccharides. P. mirabilis O33 and O49 LPSs did not induce changes in the membrane cytoskeleton, osmotic fragility and lipid peroxidation of erythrocytes. On the other hand a rise in the content of carbonyl compounds was observed for the highest concentrations of O33 LPS. This result indicated protein oxidation in the erythrocyte membrane. Lipid A, the hydrophobic part of LPS, did not change the membrane lipid fluidity and osmotic fragility of erythrocytes. Smooth and rough forms of P. mirabilis LPSs were tested for their abilities for complement-mediated immunohemolysis of erythrocytes. Only one out of seven LPSs used was a potent agent of complement-mediated hemolysis. It was rough, Ra-type of P. mirabilis R110 LPS. The O-polysaccharide-dependent scheme of reaction is presented.     

Alterations of erythrocyte structure and cellular susceptibility in patients with chronic renal failure: Effect of haemodialysis and oxidative stress

The aim of this study was to investigate erythrocytes rheological behaviour, membrane dynamics and erythrocytes susceptibility to disintegration upon strong oxidative stress induced by dialysis or by external H₂O₂ among patients with CRF. EPR spectrometry was used to investigate alterations in physical state of cellular components. Generated ROS production induced: (1) significant increase of membrane fluidity in CRF erythrocytes treated with H₂O₂ (p<0.005) and at 60 min of haemodialysis (p<0.05), (2) significant decrease of cytoskeletal protein–protein interactions (p<0.005) and (3) cellular osmotic fragility (p<0.0005). H₂O₂ exacerbated these changes. Erythrocytes from CRF patients have changed rheological behaviour and present higher susceptibility to disintegration. Erythrocytes membrane characteristics indicate that CRF patients possess younger and more flexible cells, which are more susceptible to oxidative stress. This may contribute to the shortened survival of young erythrocytes in CRF patients.     

Estimation of cell membrane properties and erythrocyte red-ox balance in patients with metabolic syndrome

Metabolic syndrome (MS) is associated with occurrence of the many cardiovascular risk factors such as atherogenic dyslipidemia, visceral fat distribution, arterial hypertension and pro-thrombotic and pro-inflammatory status. In our study the effect of disorders that appear in MS on red-ox balance and erythrocyte cell membrane properties were estimated. The study comprised 50 patients with diagnosed MS and in 25 healthy subjects. Content of thiobarbituric acid reactive substances (TBARS) and catalase, superoxide dismutase and glutathione peroxidase activity were estimated in red blood cells. Moreover, conformation status of membrane proteins, membrane fluidity and osmotic fragility were evaluated. MS was found to manifest: (1) the increase of the concentration of TBARS in erythrocytes with no statistically significant differences in antioxidant enzymes activity, (2) disorders in the structure of erythrocyte cytoskeleton proteins, (3) the increase in membrane lipids fluidity at the depth of 5th and 12th carbon atom of fatty acid hydrocarbon chain and significantly decreased fluidity at the depth of 16th carbon atom, (4) increased erythrocyte osmotic fragility.     

Reactive effect of low intensity He-Ne laser upon damaged ultrastructure of human erythrocyte membrane in Fenton system by atomic force microscopy

To find out the mechanism of modulating the deformability of erythrocytes with low intensity He-Ne laser action, we studied the effect of low intensity He-Ne laser on the ultrastructure of human erythrocyte membrane. Erythrocytes were treated with free radicals from a Fenton reaction system before exposing them to low intensity He-Ne laser. The ultrastructure of damaged erythrocyte membrane was examined by atomic force microscopy. The results showed that the erythrocyte membrane became very rough and the molecules on the surface of the membrane congregated into particles of different magnitudes sizes after treating with free radicals. Comparing the degree of congregation of the molecular particles in the non-irradiated group and the He-Ne laser irradiated （9 mW and 18 mW） group, we found the average size of molecular particles in the laser irradiated group was smaller than that in the non-irradiated group, indicating that the low intensity laser had repairing function to the damage of erythrocyte membrane produced by the free radicals.     

Improvement of stored turkey semen quality as a result of He-Ne laser irradiation

Two experiments were carried out to evaluate the effects of He-Ne laser irradiation at various energy doses on the quality of stored turkey semen. Four semen pools were used in Experiment 1. Each pool was divided into 10 aliquots, nine of which were irradiated with energy doses ranging from 0.144 to 10.8 J/cm2 while the tenth one was not irradiated (control). Each sample was evaluated for motility immediately after irradiation, 24 and 48 h later. Energy doses ranging from 3.24 to 5.4 J/cm2 had higher (P <0.01) sperm motility index (SMI) value compared to the control and samples irradiated with lower and higher laser doses. The energy dose of 3.96 J/cm2 was selected for Experiment 2 to obtain further insight on its effects on turkey sperm preservation for up to 60 h. Each pool of four semen was divided into two aliquots: one represented the control and the other one was irradiated with He-Ne laser at an energy dose of 3.96 J/cm2. Each sample was evaluated for motility and viability immediately after irradiation and then at 12 h intervals up to 60 h. The cell energy charge was also measured by HPLC. Exposure to 3.96 J/cm2 increased the SMI and viability of turkey semen stored for 60 h compared to the control (P <0.05). The cell energy charge of irradiated samples was 200% higher than in the control. Laser irradiation increased the longevity of stored turkey spermatozoa, and might be a useful technique to enhance semen quality in long-term storage.