小鼠肝枯否细胞的分离、培养与鉴定

目的研究BALB/c鼠肝枯否细胞分离、培养和鉴定。方法选用健康雌性8￣10w龄鼠,麻醉无菌取肝,D-Hank’s液注射冲洗去血并剪去部分结缔组织,剪碎肝组织,加入链霉蛋白酶E和I型胶原酶,水浴振荡消化,再加入DNaseI,共同消化。不锈钢筛网滤过,用0.005%DNaseⅠ的1640液清洗细胞液,加Tris-NH4CL溶解红细胞。30%￣70%Percoll梯度离心,用含10?S的RPMI-1640贴壁培养4￣6h洗去非贴壁细胞。通过吞噬墨水实验鉴定枯否细胞。结果枯否细胞的数量为(1.07±0.87)×107/g,贴壁率40.3%,用0.4%台盼蓝染色鉴定,细胞存活率在95%以上,吞噬实验发现90%的贴壁细胞具有吞噬功能,即KC的纯度可达90%。贴壁的枯否细胞的形态多样,已经完全伸出了伪足并且伸展贴壁,呈多边形、多角形或者梭形等不规则形状,并且胞浆内可见吞噬的微粒状物质。结论酶消化水浴振荡法结合梯度离心和贴壁培养是分离BALB/c鼠枯否细胞的可靠方法,为进一步实验打下了基础。     

应用联合酶建立BALB/c鼠枯否氏细胞体外分离培养方法

目的用作用功效不同的酶联合消化分离BALB/c鼠枯否氏细胞(KC),建立简便易行,产量、活性、纯度较稳定的KC分离培养方法。方法将0.1%型胶原酶、0.2%链霉蛋白酶、0.01%Dnase酶按1∶1∶1比例混合原位灌注和离体消化肝组织,经Percoll梯度离心,贴壁培养纯化细胞,应用荧光显微镜观察、免疫组织化学染色、吞噬功能实验进行鉴定。结果KC获得率为(2~3)×106/鼠肝,细胞存活率达96%,免疫组化和吞噬实验鉴定细胞纯度达92%。KC贴壁后形态大小不规则,呈多角形或星形,免疫组织化学染色显示溶菌酶阳性,胞浆内见吞噬的碳素墨汁颗粒。结论用作用功效不同的酶联合消化KC,经梯度离心和贴壁培养,可获得高产量、高活性、高纯度的BALB/c鼠KC,为进一步研究KC在肝脏疾病中的作用提供可靠的细胞来源。     

小鼠肝枯否细胞分离培养与免疫组化鉴定

门静脉灌注ＧＢＳＳ以清除肝内淤血，链霉蛋白酶Ｅ选择性破坏肝脏实质细胞，可以获得肝间质细胞悬液。经Ｐｅｒｏｃｌｌ梯度离心，贴壁养得到大量纯化枯否细胞，数量大约为４．１＊１０＾６个／克肝组织。     

大鼠肝星形细胞、枯否细胞的同步分离培养与鉴定

目的:建立经济、简便的同步分离培养肝星形细胞及枯否细胞的方法。方法:应用胶原酶和蛋白酶对大鼠肝脏进行原位灌流、消化,获得大鼠肝脏非实质细胞,经18%(W/V)的Nycodenz密度梯度离心,一步分离肝星形细胞及枯否细胞;肝星形细胞处在界面,枯否细胞则处在界面下;台盼蓝染色鉴定细胞活力;Desmin免疫细胞化学染色鉴定肝星形细胞;溶菌酶免疫细胞化学染色鉴定枯否细胞。结果:该方法分离的肝星形细胞和枯否细胞的纯度达95%左右。肝星形细胞得率(3.0±0.5)×107/肝,枯否细胞得率(4.0±0.5)×106/肝。结论:应用Nycodenz一步密度梯度离心法可以很好的同时分离肝星形细胞和枯否细胞。     

SD大鼠肝脏枯否细胞分离方法改进研究

目的：研究一种简单、高效、实用并且稳定的SD大鼠肝脏原代枯否细胞( KCs)的提取及培养方法。方法选择体质量200 g左右的健康SD雄性大鼠,采用0．05% IV型胶原酶在体原位灌注法结合剪碎法,37℃水浴振荡20 min,经30%和60%Percoll密度梯度离心后,再经40% Percoll离心及贴壁法纯化提取KCs。倒置显微镜下观察细胞形态,并用台盼蓝染色实验鉴定细胞活力,吞墨实验观察细胞吞噬功能及流式细胞术鉴定细胞纯度。结果每只大鼠平均肝脏KCs的获得量为(1．00±0．20)×107(n=20),细胞活力为(92．34±0．15)%,细胞纯度均在(93．56±0．24)%。结论改良的SD大鼠肝脏KCs提取方法操作简单,效果可靠,可为今后KCs的研究奠定基础。     

肝清除功能与肝切除后残肝损伤的关系

利用肝切除（６８％），肝切除＋ＬＰＳ大鼠，观察枯否细胞吞噬、肝清除功能与肝损伤之关系。结果发现在肝切除后果早期枯否细胞吞噬及肝清除明显受抑，残余肝出现病理改变。肝清除持续低下至术后７天渐趋恢复，肝损伤病理计分结果与之相平等。当肝切除动物 外源性ＬＰＳ攻击时，枯否细胞吞噬上清除降低更为显著，同时肝损伤亦随之加重，结果提示在肝切除病人中适当减少切除量，保护枯否细胞，以及防止肠原性感染具有重要意义。     

鼠肝枯否细胞的分离,培养和鉴定

目的：研究鼠肝Ｋｕｐｆｅｒ细胞的分离、培养和鉴定。方法：用链酶蛋白酶和胶原酶原位灌流，ｎｙｃｏｄｅｎｚ密度梯度离心分离大鼠Ｋｕｐｐｆｅｒ细胞，再经贴壁培养，并应用免疫组织化学、吞噬功能试验，电镜等方法进行鉴定。结果：本法能成功地获得高纯度的Ｋｕｐｆｅｒ细胞，Ｋｕｐｆｅｒ细胞得率为３～５×１０６／肝，贴壁后呈典型的星形及多角形，免疫组化染色示ｌｙｓｏｚｙｍｅ阳性，胸浆内见吞噬的ｉｎｄｉａｉｎｋ及ｌａｔｅｘｂｅａｄｓ颗粒，电镜观察细胞表面有发达的伪足、微绒毛，胞浆内含大量溶酶体及吞噬ｌａｔｅｘｂｅａｄｓ颗粒。结论：本实验所用的Ｋｕｐｆｅｒ细胞分离培养方法，简单易行，可靠，细胞纯度高，可用于进一步研究Ｋｕｐｆｅｒ细胞的生物学功能     

二羧乙基锗倍半氧化物（Ge－132）对肝枯否细胞免疫及酶活性的影响

探讨了二羧乙基锗倍半氧化物（Ｇｅ－１３２）对肝枯否细胞免疫及酶活性的影响，二羧乙基锗倍半氧化物在体外实验中明显地对抗四氯化碳对大鼠肝枯否细胞的损害、明显增加花环形成率、吞噬率及吞噬指数，而且显著增加过氧化物小体、过氧化物酶、琥珀酸脱氢酶活性．实验结果表明，四氯化碳可以损伤肝枯否细胞，二羧乙基锗倍半氧化物（Ｇｅ－１３２）可保护和修补四氯化碳的细胞损伤．     

光动力疗法诱导Hela细胞凋亡的实验研究

1 一般资料 1．1准备工作：细胞复苏、培养、传代、冻存：人宫颈癌细胞系Hela为贴壁细胞，从液氮中取出冻存的细胞后立即置于37℃水浴中融解。1000r／min离心5分钟，弃上清后加入RP—M11640培养基离心洗涤1次。加入含10％小牛血清的RP—M11640培养基5ml悬浮起细胞后置于25ml细胞培养瓶中，     

草苁蓉对损伤肝枯否细胞免疫活性的影响

将Ｗｉｓｔａｒ系大鼠乳鼠肝枯否细胞体外培养，选生长良好的细胞培养瓶分为正常组、损伤组、实验组及阳性对照组，除正常组外均以四氯化碳损伤细胞，实验组再加草苁蓉乙醇提取物，阳性对照组加ＰＨＡ。然后在各培养瓶中加入已吸附兔抗绵羊红细胞抗体ＩｇＧ和ＳＲｂＣ，做花环形成实验和吞噬试验，镜下计数并算出枯否细胞在特异抗体介导下的花环形成率、蚕噬率及吞噬指数。结果，实验组比损伤组均显著增（Ｐ＜０．０１）。     

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.     

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.     

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…     

安心颗粒对急诊经皮冠状动脉介入术后生活质量影响的研究

目的：评价使用安心颗粒对急诊经皮冠状动脉介入术（PPCI）术后生活质量的影响.方法：将160例接受PPCI的急性ST段抬高型心肌梗死患者随机分为安心颗粒组（术前顿服安心颗粒8.8g,术后安心颗粒4.4 g/次,每日2次）和对照组（仅接受基础药物治疗）.所有患者均服用阿司匹林、氯吡格雷和阿托伐他汀.分别在入院时、出院前1d、出院后180 d时,应用心肌梗死多维度量表（MIDAS）、中文版SF-36评价量表对患者生活质量评分.并观察术后30 d以内的出血并发症、血小板减少症发生情况.结果：入院时和出院前1d,两组患者的心肌梗死MIDAS、SF-36量表评分比较无差异（P＞0.05）;出院后180 d时,与对照组比较,安心颗粒组MIDAS、SF-36评分明显减低（P＜0.05）;组内与入院时比较,两组出院前1d、出院后180 d时,MIDAS、SF-36评分均降低（P＜0.05）.两组患者在随访期间均无大量出血、少量出血、重度和极重度血小板减少症发生,安心颗粒组有4例、对照组有7例发生不明显出血（P＞0.05）.两组发生轻度血小板减少症的患者数比较无差异（P＞0.05）.结论：PPCI使用安心颗粒,能改善急性ST段抬高型心肌梗死患者的生活质量,且不增加出血风险.     

The comparative studies of the influences of Urapidil and Nicardipine on sino-atrial node function, atrio-ventricular node function and hemodynamics

Objective:To investigate the influences of urapidil and nicardipine on rabbit sinus function,atrio-ventricular node function and hemodynamics.Methods:Thirty-two Angora's rabbits were selected and randomly divided into four groups.U1 group:urapidil 0.25 mg/kg;U2 group:urapidil 0.5 mg/kg;N1 group:nicardipine 10 μg/kg;N2 group:nicardipine 20 μg/kg.All these medicine were administrated within 30 seconds.Measurements were taken before and after the administration of urapidil or nicardipine for the following data:mean blood pressure(MAP),heart rate(HR),sino-atrial conduction time(SACT),maximal sinoatrial recovery time(SNRTmax)corrected sinus node recovery time(CSNRT),index of sinus node recovery time(SNRTI),Wenckebach A-V conduction frequency (WB),and P-R interval.Results:Significant MAP and HR changes were identified in all of the four groups before and after administration of both urapidil and nicardipine.No significant changes could be found in the rest of the parameters.Intergroup analysis showed that SACT and CSNRT of N1 and N2 groups were shorter than those of the U2 group(P＜0.01);the MAP decreased(P＜0.01)and the HR increased drastically(P＜0.01).Conclusions:Neither urapidil(0.25 mg/kg,0.5 mg/kg)nor nicardipine(10μg/kg,20μg/kg)has any significant influence on rabbit sinus function or rabbit atrio-ventricular node function.Nicardipine could be a better choice than urapidil for parafunctional sinus node patients.     

Dynamic Changes in Serum Estradiol and Progesterone levels in Patients of Premenstrual Syndrome with Adverse Flow of Liver-qi

Objective:To probe into the influence of changes of ovarian hormones on the pathogenesis of the specific sub-type premenstrual syndrome(PMS)and reveal partial microcosmic mechanisms of adverse flow of liver-qi.Methods:Estradiol(E2)and progesterone(P)levels in serum were determined at different phases of menstrual cycle by radioimmunoassay.Results:In the group of PMS with adverse flow of liver-qi.the secretive peak value Of E2 and P at the follicular phase significantly decreased,and the secretive peak value at the luteal phase did not come into being.Conclusions:Low E2 and P secretive peak at the follicular phase and absence of secretive peak at the luteal phase is one of the microcosmic mechanisms of PMS with adverse flow of liver-qi.One of the pathophysiologic mechanisms of specific sub-type PMS is probably the continuous low level of E2and P.     

Real-time Three-dimensional Echocardiography in Assessment of Left Ventricular and Right Ventricular Volumes

Real-time three-dimensional echocardiography (RT3DE)is a new ultrasound technique that enables dynamic threedimensional visualization and quantification of the heart in real time. Investigation of feasibility and methodology of RT3DE in determining left ventricular (LV) and right ventricular (RV) volumes, RT3DE was performed in 35 normal adults using Philips SONOS 7500 system with a 2-4 MHz matrix array transducer. The 60°×60° "pyramid" volume database was obtained and analyzed on a TomTec echo workstation. Both LV and RV volumes were calculated with four 3DE methods (i.e. apical 2, 4, 8, and 16-plane) through manually tracing ventricular endocardial borders in end diastole and end systole. Stroke volumes were then calculated. LV volume was also measured by 2DE Simpson's rule using GE VIVID 7 ultrasound machine.     

SCREENING FOR A 21-CHROMOSOME ABNORMALITY IN PREIMPLANTED EMBRYOS OF ELDERLY WOMEN

Increasing maternal age is the only etiological factor unequivocally linked to Down's syndrome in humans. The occurrence rate of newborns with Down's syndrome is about 1/220 in women over 35 years old. However, the occurrence rate in embryos fertilized in vitro, of the elder woman is unclear. Using FISH we screened the number of chromosome 21 in preimplanted embryos of 5 elderly women (average age, 38.4 years) to study the feasibility and necessity of screening trisomy 21 in embryos in patients over 35 years old at the in vitro fertilization (IVF) center.     

Scientific principles and rigorous processes should be followed in developing clinical guidelines for therapeutic interventions of integrative medicine

A clinical guideline for the therapeutic interventions of integrative medicine may be defined as a written document which states a series of recommendations on therapeutic interventions of integrative medicine for a special disease or condition. The guideline may provide assistance to medical professionals in making clinical decisions aimed at improving the clinical outcome of patients and reducing the costs of medical care（~＇4~. Recommendations issued by a guideline should be based on the best available evidence in both Western and Chinese medicine. For fulfilling this purpose, the development of clinical guidelines for therapeutic interventions in the field of integrative medicine should follow scientific principles and undergo a rigorous processes.