离体肺组织切片的制备和孵育

目的建立一种简便有效的离体肺组织切片孵育方法.方法利用肺组织精细切片技术制备肺组织切片,以0.5 mL Krebs-Henseleit(K-H)缓冲液作为孵育液,在37 ℃培养箱内分别孵育1、2、3、4 h,测定肺片的四甲基偶氮唑盐(MTT)比色、乳酸脱氢酶(LDH)释放率和三磷酸腺苷(ATP)含量,以反映肺组织活性.结果肺片在37 ℃培养箱内孵育1、2、3、4 h后MTT比色、LDH释放率和ATP含量均无统计学差异(P>0.05).结论应用0.5 mL Krebs-Henseleit(K-H)缓冲液作为孵育液、在37 ℃培养箱对肺片进行孵育时组织活性没有明显变化,可作为一种简便有效的孵育方法用于离体肺片研究.     

过氧化氢对大鼠离体肺组织精细切片活性的影响

目的研究过氧化氢(H2O2)对离体肺组织精细切片活性的影响,建立肺组织氧化损伤的简易模型。方法制备离体肺组织精细切片,以0.5mLKrebsHenseleit缓冲液作为孵育液,在37℃培养箱内进行孵育。在孵育液中加入H2O2,使其浓度为2mmol/L,分别孵育0(对照组)、5、10、15和20min;然后在不含H2O2的孵育液中进行恢复孵育1h。测定肺片的四甲基偶氮唑盐(MTT)染色、孵育液中乳酸脱氢酶(LDH)和肺片组织ATP含量。结果随H2O2损伤时间延长,MTT染色变浅,其吸光度逐渐减小,损伤时间大于10min组与对照组比较有显著差异(P<0.01)。H2O2损伤10min组肺片的LDH释放多于对照组(P<0.05),损伤时间更长者增加更明显(P<0.01)。H2O2损伤5min组的肺片组织中ATP含量明显低于对照组(P<0.05),损伤时间更长者下降更明显(P<0.01)。经偏相关分析发现,肺片MTT染色与LDH释放呈负相关(r=-0.866,P<0.01),与ATP含量呈正相关(r=0.869,P<0.01)。结论离体肺组织切片在含H2O2的孵育液中进行孵育时,随孵育时间延长其活性降低。此模型简单可靠,可用于肺组织氧化损伤的研究。     

过氧化氢对大鼠离体肺组织精细切片活性的影响

目的研究过氧化氢(H2O2)对离体肺组织精细切片活性的影响,建立肺组织氧化损伤的简易模型.方法制备离体肺组织精细切片,以0.5 mL Krebs-Henseleit缓冲液作为孵育液,在37℃培养箱内进行孵育.在孵育液中加人H2O2,使其浓度为2 mmol/L,分别孵育0(对照组)、5、10、15和20 min;然后在不含H2O2的孵育液中进行恢复孵育1 h.测定肺片的四甲基偶氮唑盐(MTT)染色、孵育液中乳酸脱氢酶(LDH)和肺片组织ATP含量.结果随H2O2损伤时间延长,MTT染色变浅,其吸光度逐渐减小,损伤时间大于10 min组与对照组比较有显著差异(P＜0.01).H2 O2损伤10 min组肺片的LDH释放多于对照组(P＜0.05),损伤时间更长者增加更明显(P＜0.01).H2O2损伤5 min组的肺片组织中ATP含量明显低于对照组(P＜0.05),损伤时间更长者下降更明显(P＜0.01).经偏相关分析发现,肺片MTT染色与LDH释放呈负相关(r=-0.866,P＜0.01),与ATP含量呈正相关(r=0.869,P＜0.01).结论离体肺组织切片在含H2O2的孵育液中进行孵育时,随孵育时间延长其活性降低.此模型简单可靠,可用于肺组织氧化损伤的研究.     

碱性成纤维细胞生长因子拮抗大鼠心肌细胞缺氧/复氧损伤的机制

目的 :研究碱性成纤维细胞生长因子 (basicfibroblastgrowthfactor ,bFGF)拮抗大鼠心肌细胞缺氧 /复氧(hypoxia/reoxygenation ,H/R)损伤的机制。 方法 :H/R处理的大鼠心肌细胞的孵育液中 ,对照组不加任何处理因素 ,其它各组分别加bFGF(10 μg·L-1)、bFGF与丝裂素活化蛋白激酶 (mitogen activatedproteinkinase,MAPK)抑制剂PD980 5 9(5 0 μmol·L-1)、蛋白激酶C(proteinkinaseC ,PKC)抑制剂H7(40 μmol·L-1)、一氧化氮合酶 (nitricoxidesynthase ,NOS)抑制剂左旋硝基精氨酸甲酯 (NG nitro L argininemethylester,L NAME ,5 0 μmol·L-1)。孵育完毕 ,测定细胞活力、ATP含量及孵育液中乳酸脱氢酶 (lactatedehydrogenase,LDH)含量。 结果 :H/R组心肌细胞活力较对照组降低 32 % (P <0 .0 1) ,细胞ATP含量减少 5 8% (P <0 .0 1) ,孵育液中LDH活性增加 5 .8倍 (P <0 .0 1) ;bFGF组心肌细胞活力较H/R组增高 17% (P <0 .0 1) ,细胞ATP含量增加 45 % (P <0 .0 1) ,孵育液中LDH活性降低 31% (P <0 .0 1) ,细胞PKC、MAPK活性分别增加 32 % (P <0 .0 1)和 10 % (P <0 .0 5 )。PD980 5 9、H7、PD980 5 9+H7及L NAME各组心肌细胞活力较bFGF组分别降低 13%、17%、2 4%和 2 7% (均P <0 .0 1) ,细胞ATP含量分别减少 2     

百日咳菌液对大鼠培养脑切片兴奋性氨基酸释放的影响

目的:了解百日咳菌液对培养的大鼠脑切片兴奋性氨基酸(EAAs) 释放的影响及11 双氢5H二苯(a ,d)环庚烯亚胺( MK801) 对脑切片有无保护作用。方法:SD 大鼠随机分成正常脑切片培养组(NC组) ,10 % 百日咳菌液脑切片培养组(PBC 组) 和MK801 预处理后10 % 百日咳菌液脑切片培养组( MPBC组) 。用高效液相色谱法测脑切片培养上清液中的EAAs 释放量。结果:PBC 组的谷氨酸(Glu) 、门冬氨酸(Asp) 及乳酸脱氢酶(LDH) 的释放量明显高于MPBC 组及NC 组。结论:百日咳菌液可使培养的脑切片组织EAAs 和LDH 的释放量增多;MK801 预处理可降低脑切片组织EAAs 及LDH 的释放量。提示MK801 对离体脑组织有保护作用     

聚乙烯吡咯烷酮防止酶扩散的观察

骤冷组织的恒冷切片如不经固定,直接孵育,可出现酶的扩散,影响酶的定位,甚至出现假阴性反应。为加深对酶扩散的后果和聚乙烯吡咯烷酮(PVP)对防止酶扩散作用的认识,作者对胞液内的可溶性酶,LDH和G-6-PD在溶液中的扩散作用和聚乙烯吡咯烷酮的防扩散效果进行了观察。 取SD大鼠肝、肾及横纹肌组织,合包骤冷后,切片分组作如下处理:Ⅰ组在缓冲液内,37℃预处理10分钟;Ⅱ组缓冲液内加入PVP     

褪黑素受体亚型蛋白的免疫组化检测方法

1　方法介绍1.1　组织标本制备　新鲜待检组织立即用 4%多聚甲醇固定 ,常规脱水、透明、浸蜡 ,石蜡包埋组织 ,4℃冰箱贮存待检测。1.2　主要步骤　将石蜡包埋组织作 4μm的连续切片 ,在 37℃烘箱过夜。石蜡切片用二甲苯脱腊 10 min× 3,梯度乙醇洗 2min× 3,3%过氧化氢溶液处理 10 m in,以阻断内源性过氧化物酶活性。蒸馏水洗 2 min× 3,0 .0 1mol/ L ,p H 6 .0柠檬酸缓冲液微波修复抗原 10 min,冷却后蒸馏水洗 2 m in× 3,PBS洗 2 m in× 3。每张切片加 5 0 ml非免疫性动物血清温室孵育 10 min,每张切片加 5 0 ml鼠抗人 mt1 和 MT2 受体…     

心肌细胞培养方法和活细胞噻唑兰法测定

目的:介绍心肌细胞培养方法和研究比色分析法测定活心肌细胞.方法:接种不同数量细胞培养后行噻唑兰(MTT)比色分析;正常培养、含不同浓度维拉帕米培养的细胞行MTT代谢比色分析,并测乳酸脱氢酶(LDH)活性.结果:比色值与接种细胞数量、细胞释放的LDH活性呈高度相关(r=0.996、-0.986,P<0.01).结论:MTT比色分析法可用于测定活心肌细胞.     

可乐定置换物对正常小鼠离体胰岛功能的影响

目的　探讨可乐定置换物 (CDS)对胰岛功能的影响。方法　胶原酶和DNA酶联合消化法分离NMRI小鼠胰岛 ,RPMI1 640组织培养液过夜培养 ,采用MilliporeMultiscreen系统观察CDS对胰岛功能的影响。放免法测定孵育液中胰岛素及胰高糖素的浓度。结果　 (1 )离体胰岛胰高糖素的分泌明显受葡萄糖浓度的抑制 ,CDS显著抑制胰高糖素的分泌 ,且抑制强度明显依赖于CDS的浓度。 (2 )离体胰岛胰岛素的分泌依赖于孵育液中的葡萄糖浓度 ,CDS明显刺激胰岛素的释放。Ca2 + 通道阻滞剂硝苯吡啶 (2 5μmol·L- 1 )及K+ 通道开放剂二氮嗪(1 0 0 μmol·L- 1 )可显著抑制胰岛素释放 ,但其作用可被CDS废除。结论　作为咪唑啉受体的内源性配体 ,CDS可刺激离体胰岛胰岛素的释放 ,抑制胰高糖素的分泌     

甘氨酸在神经母细胞瘤细胞ATP耗竭损伤中的保护作用

目的:观察甘氨酸(glycine,Gly)对ATP耗竭的神经母细胞瘤细胞SH-SY5Y的保护作用及其可能机制.方法:通过乳酸脱氢酶(LDH)释放、四甲基噻唑蓝(MTT)实验及细胞内ATP含量的检测,观察ATP耗竭时Gly对SH-SY5Y细胞的保护作用.通过Western blot检测甘氨酸受体的表达情况.结果:耗竭1 h内,Gly保护组细胞内的ATP含最高于耗竭组.耗竭6 h后.Gly保护组细胞的LDH释放率较耗竭组明显下降,MTT吸光度也明显上升.同时,ATP耗竭后细胞甘氨酸受体表达上调,这种上调趋势在使用甘氨酸后有所缓解.结论:甘氨酸町保护ATP耗竭的SH-SY5Y细胞,并缓解细胞应激时甘氨酸受体的表达上调.     

Value of D-Dimers in patients with acute aortic dissection

Objective: To evaluatel the value of D-dimers in patients with acute aortic dissection (AAD). Methods: This study consisted of 16 patients with AAD and 27 non-AAD patients. Serum D-dimets were measured by Sta-Liatest D-DI immunoturbidimetric assay. Results: D-dimer level was higher (P ＜ 0.001) in patients with AAD(7.91 ± 5.52 μg/ml) than that in non- AAD group(1.57±1.24 μg/ml). D-dimer was positive (＞0.4 μg/ml) in all patients with AAD and in 10 control group patients (37%). Among patients with acute AAD, D-dimers tended to be higher in Stanford A than in Stanford B (8.67 ± 4.31 μg/ml vs. 3.24±1.27 μg/ml, P ＜0.01). D-dimer values tended to be higher in more extended disease(3.84 ± 1.65 μg/ml, 8.57 ± 3.58 μg/ml and 11.87 ± 5.69 μg/ml in thoracic aorta, thoracic and abdominal aorta, thoracic and abdominal aorta and iliacal arteries, respectively, P ＜ 0.05 for both 8.57 ± 3.58 and 11.87 ± 5.69 vs. 3.84 ± 1.65 ). Including the control group into the analysis, we found a sensitivity of 100%, a negative predictive value of 100%, and a specificity of 66% and a positive predictive value of 64% for D-dimer in diagnosis of AAD in our patients with suspected AAD. Conclusion: D-dimer was elevated in patients with AAD. A negative D-dimer test result could be useful in excluding AAD.     

Research of combined liver-kidney transplantation model in rats

Objective： To set up a simple and reliable rat model of combined liver-kidney transplantation. Methods： SD rats served as both donors and recipients. 4℃ sodium lactate Ringer＇s was infused from portal veins to donated livers,and from abdominal aorta to donated kidneys, respectively. Anastomosis of the portal vein and the inferior vena cava （IVC） inferior to the right kidney between the graft and the recipient was performed by a double cuff method, then the superior hepatic vena cava with suture. A patch of donated renal artery was anastomosed to the recipient abdominal aorta. The urethra and bile duct were reconstructed with a simple inside bracket. Results： Among 65 cases of combined liver-kidney transplantation, the success rate in the late 40 cases was 77.5%. The function of the grafted liver and kidney remained normal. Conclusion： This rat model of combined liver-kidney transplantation can be established in common laboratory conditions with high success rate and meet the needs of renal transplantation experiment.     

APPLICATION OF LORNOXICAM TO PATIENT-CONTROLLED ANALGESIA IN PATIENTS UNDERGOING ABDOMINAL SURGERIES

FOR anesthesiologis s ,treatingpostoperativepainhas alwaysbeen a problem.Althoughopioidshave been provedtobe effective,theirsideeffectscouldnotbeignored.With thedevelopmentofscienceand pharmacology,many drugs with aspectsof satisfactoryanalgesicefficacyand couldbe welltoleratedby patientshave been developed.And lornoxicamisone of them, which isa non-steroidalanti-inflammatorydrug (NSAID ), with analgesic, anti-infl-ammatory,andantipyreticproperties.Itseliminationhalf-time(3 to 5 hours) isle…     

BLOOD PRESSURE CHANGE WITH AGE IN SALT-SENSITIVE TEENAGERS

Objective To observe blood pressure change with age in salt-sensitive teenagers whose salt sensitivity were determined by repeated testing.Methods Salt sensitivity was determined through intravenous infusion of normal saline combined with volume-depletion by oral diuretic furosemide in 55 teenagers. After five years, salt sensitivity was re-examined and subject blood pressure was followed up. Blood pressure changes in salt-sensitive teenagers were compared to that of non-salt sensitive teenagers over five years.Results After 5 years, the repetition rate of salt sensitivity determined by intravenous saline loading is 92.7%. In teenagers with salt sensitivity on the baseline, both the systolic blood pressure increments and increment rates were much higher than non-salt sensitive teenagers (12.7±12.1 mmHg vs. 2.8±5.2 mmHg, P＜ 0.01; 12.2%± 12.0% vs. 2.5% ±4.4%, P＜ 0.001,respectively). There was a similar trend for diastolic blood pressure (8.4 ± 6.4 mmHg vs. 3.7 ± 6.4 mmHg, P = 0.052; 13.2% ±10.6 % vs. 6.8%± 10.1%, P = 0.053, respectively).Conclusions Salt sensitivity determined by intravenous saline loading showed good reproducibility. Blood pressure increments with age were much higher in salt-sensitive teenagers than non-salt sensitive teenagers, especially in terms of systolic blood pressure.     

Development on Adjustable Calibration Marker for Shock Wave Focus

Shock wave lithotripsy （SWL） is a treatment of choice for upper urinary stones. However, this procedure is inappropriate for obese patients because the focus is often unable to reach the target owing to the limited focal distance in shock wave source. Although treating such patients in a blast path may increase the application length of shock wave source, it＇s difficult to find this path on the lithotripter monitor. For this reason, we invented an adjustable calibration marker in order to set an effective focus in the shock wave hath.     

Study on the TCM Pathogenesis of Angiopathic Complications of Type II Diabetes

Excess production of reactive oxygen species(ROS)of mitochondrion mediated by hyperglycemia is the common pathogenesis of angiopathic complications of diabetes.TCM holds that the damp from the dysfunction of spleen.kidney and liver is the causative factor of complications of diabetes.This is similar to the mechanism of Ros resulting in angiopathic complications of diabetes.When the angiopathic complications of type II diabetes mellitus(T2DM)are difierentiated as caused by turbid damp in TCM can be explained as ROS.Since the obstruction of pathogenic damp in channels and collaterals is said to be the main pathogenesis,the treating principle should be dissolving the damp to remove the obstruction.     

Leptin, Ghrelin and TNF-α before and after Hypo-caloric Traditional Chinese Diet and Auricular Acupuncture

INTRODUCTION Obesity is a complex emergent problem, which can be possibly solved not only by the diet but also by the life style and promotion of a constant physical exercise. 1, 2 No doubt careful attentions must be given to the nutritional condition of obese people, the dietary habits, the somatic build (i.e. distribution of fat mass) and the organic functions linked to formation of the fat mass. All the parameters should be constantly monitored before, during and after a diet treatment. 3, 4, 5     

Relationship between Dysglycemia and Carotid Atherosclerosis in Tibetan Population

People with dysglycemia are at high risk for atherosclerotic diseases. This study aims at investigating the atherosclerotic vascular damage in dysglycemia and its metabolic origin in Tibetan population.     

Acupuncture Combined with Spinal Tui Na for Treatment of Primary Dysmenorrhea in 30 Cases

Objective: To observe the therapeutic effects in acupunture treatment of primary dysmenorrhea combined with spinal Tui Na, and study its mechanism. Methods: Thirty cases of the treatment group were treated by acupuncture combined with spinal Tui Na, and thirty cases in the control group were treated by routine acupuncture. Results: The total effective rate was 93.3% in the treatment group, and 73.3% in the control group, with a significant difference between the two groups (P<0.05). Conclusions: Acupuncture combined with spinal Tui Na has good prospects for treatment of primary dysmenorrhea.     

Luo Lingjie's Experience in Treating Chronic Nephropathy

In treating chronic nephropathy,Luo Lingjie,a chief physician,pays attention to regulating the balance between yin and yang,treating infection if present,and removing pathogenic factors.He prescribes gentle drugs and uses carefully strongly warming-tonifying ones,emphasizes the importance of persuading the patient to persist in treatment with medication and nurse one's health for recuperation,and is good at combined use of TCM and western medicine therapy and brings the merits of various therapies into full play,with obvious theraoeutic effects.