小鼠囊胚细胞DNA损伤与维甲酸的诱导作用

目的观察维甲酸(RA)对小鼠囊胚细胞DNA的损伤,进一步证实RA对囊胚细胞凋亡效应。方法获取妊娠3.5d小鼠囊胚,分别培养在含0、1μmol/l和10μmol/l的RA的M199培养基中24h,用单细胞凝胶电泳技术检测RA对体外培养小鼠囊胚细胞DNA的损伤。结果 RA可破坏囊胚细胞DNA的结构,诱导细胞凋亡,导致囊胚滋养层和内细胞群细胞减少。结论维生素A酸对小鼠胚胎的生长发育具有细胞毒性作用。     

不同饲养层细胞对胚胎干细胞和内细胞团的影响

目的探讨不同饲养层细胞对小鼠胚胎干细胞（ESC）和兔囊胚内细胞团（ICM）的影响。方法分别用成系小鼠胚胎成纤维细胞（SNL）、兔胚胎成纤维细胞（REF）、兔肾脏成纤维细胞（RRF）作为饲养层饲养小鼠ESC和兔囊胚，观察它们的生长状态，并分别检测其碱性磷酸酶活性。结果生长在REF上的小鼠ESC具有最显著的未分化形态，保持未分化时间最长．碱性磷酸酶活性最强。在REF上呈聚集生长的兔囊胚ICM的比例最高。结论REF能维持小鼠胚胎干细胞的未分化状态及促进兔囊胚ICM聚集生长，可以作为ESC的饲养层细胞。     

用饲养层分离胚胎干细胞集落的实验初探

目的 用饲养层分离胚胎干细胞集落。方法 用胚龄为13~14 d的小鼠胚胎分离原代成纤维细胞，制成饲养层，用于囊胚的培养。结果 小鼠原代胚胎成纤维细胞（PMEF）贴壁能力较好，增殖快，易铺层。囊胚和内细胞团（ICM）在饲养层上贴壁生长良好，当培养4~5 d时，其增殖率为16/28（57%）。在ICM离散48 h后，各种胚胎干细胞（ES）集落开始出现。此种集落经碱性磷酸酶染色成阳性。结论 用饲养层分离胚胎干细胞获得初步成功。     

应用卵裂期废弃胚胎培养建立人胚胎干细胞系的方法探索

目的：探讨应用卵裂期废弃胚胎,采用两种不同培养法建立人胚胎干细胞（hESC）系的方法。方法：将临床IVF及ICSI周期中剩余废弃的卵裂期胚胎继续培养至扩张囊胚,分别用免疫切割法及全囊胚培养法分离出内细胞团（ICM）,接种在用丝裂霉素处理过的小鼠胚胎成纤维细胞饲养层上,用hESC培养液培养、传代,建立胚胎干细胞系。通过胚胎干细胞生物学特性、特异性表面标志物、核型分析及体内外分化全能性等传统方法进行鉴定。结果：132枚废弃的卵裂期胚胎在体外继续培养后,有28枚发育为囊胚,随机分组14枚囊胚采用免疫切割法分离ICM,未得到干细胞系;另14枚采用全囊胚培养法,最终得到1个干细胞系,经鉴定,这些细胞都具有hESC的生物学特性。结论：①部分废弃卵裂期胚胎能够发育至囊胚期,可作为建立hESC系的材料来源之一;②全囊胚培养法建系成功率高于免疫切割法,尤其适用于评分低的囊胚。     

胰岛素对不同时期ICR小鼠胚胎体外发育的影响

目的研究胰岛素对不同时期小鼠胚胎体外发育的影响.方法收集1-细胞、2-细胞、4-细胞及桑椹胚期鼠胚,分别在含0（对照组）、0.005、0.05、0.5、5、10μg/mL胰岛素的KSOM中培养.结果在1-细胞及2-细胞阶段添加系列浓度时,0.005μg/mL及0.05μg/mL胰岛素组不仅提高囊胚率、孵化率,而且提高囊胚细胞数.4-细胞阶段添加系列浓度时0.05μg/ml胰岛素组能够提高囊胚率、孵化率以及囊胚细胞数;而至桑椹胚阶段再添加系列浓度胰岛素时,囊胚率、孵化率以及囊胚细胞数均无统计学意义（P〉0.05）.结论在ICR小鼠胚胎体外培养时适宜的胰岛素浓度为0.05μg/mL;桑椹胚之前添加胰岛素对小鼠胚胎发育是必要的.在一定发育阶段内,随着胚胎的生长对胰岛素的需求也相应增加.     

小鼠胚胎干细胞建系及GFP标记

目的：为组织工程提供新型种子细胞,建立胚胎干（ES）细胞系。方法：取129／svj白色小鼠4.5d的囊胚,分离囊胚的内细胞团（ICM）细胞;在条件培养液及小鼠胚胎成纤维细胞（MEF）作铺层下,细胞扩增传代,碱性磷酸酶（AKP）染色及oct4、ssea-1等特异性抗体鉴定细胞,并将细胞注射至裸鼠皮下观察细胞在动物体内的生长变化。结果：细胞体外培养呈“巢状”生长,可以持续传代;AKP染色及oct4、ssea-1等特异性抗体免疫反应均阳性;裸鼠皮下注射的细胞发育生长为包含各胚层细胞成分的畸胎瘤。在此基础上以基因转染方法将细胞标记绿色荧光蛋白（GFP）。结论：从小鼠ICM所分离的细胞为小鼠胚胎干细胞,且标记GFP的ES细胞在传代及形成胚体的过程中均能显示GFP,对将来组织构建的种子细胞起到示踪作用。     

由囊胚内细胞团分离人胚胎干细胞

胚胎干（Embryonic stem，ES）细胞是由着床前胚胎内细胞团（Inner cell mass，ICM）或原始生殖细胞（Primordial germ cells，PGC）经体外分离培养而建立的克隆细胞系。ES细胞的特点是只生长、不分化，并保持早期胚胎发育的多能性。本文比较了人囊胚在G２．２培养液和含DMEM培     

小鼠胚胎成纤维细胞与人胚共培养对胚胎体外发育影响

[目的]评价小鼠胚胎成纤维细胞共培养是否促进d3质量差的人胚胎的囊胚发育。[方法]使用小鼠胚胎成纤维细胞与受精后第3天(d 3)质量差的胚胎共培养,对照组使用Quins囊胚培养液进行序贯培养,观察各阶段囊胚形成率,检测囊胚总细胞数。[结果]d5共培养组12.3％胚胎开始出现早期囊腔化,d 6 7.4％胚胎出现囊腔扩张,4.2％的胚胎d 7孵出。而序贯培养组d 5仅4.2％胚胎开始早期囊腔化,d 6仅1.5％胚胎出现囊腔扩张,d 7仅0.6％的胚胎孵出,表明共培养组囊胚形成率更高,囊胚发育速度快于对照组。此外,共培养组囊胚总细胞数更多,d 6扩张囊胚总细胞数为62.6±13.4,而未扩张囊胚总细胞数26.1±8.9,分别高于序贯培养组。[结论]小鼠胚胎成纤维细胞共培养与 Quins囊胚培养液序贯培养比较,前者更能促进d 3质量差的人胚胎体外发育。     

子宫内膜上皮细胞与小鼠胚胎共培养对小鼠胚胎早期发育的影响

目的:采用子宫内膜上皮细胞与小鼠胚胎共培养,观察其对早期胚胎体外发育的影响。方法:对小鼠子宫内膜上皮细胞进行分离、培养鉴定,然后与体外受精的小鼠胚胎进行共培养,观察共培养对小鼠胚胎卵裂率、孵化率以及囊胚期胚胎总细胞数的影响。结果:与无上皮细胞相比,共培养明显促进了小鼠桑椹胚率(71.3%和61.2%,P<0.05),囊胚率(50.0%和39.7%,P<0.05),囊胚孵化率(34.8%和24.3%,P<0.05)和胚胎总细胞数(44.1和35.6,P<0.05)。结论:子宫内膜上皮细胞能够促进小鼠胚胎的体外发育,对早期胚胎的体外发育环境具有优化作用。     

用低形态学评分D3胚胎建立人胚胎干细胞系

寻找人胚胎干细胞（hESC）建系材料来源。选用体外受精（IVF）低形态学评分的D3胚胎行序贯囊胚培养，用免疫外科的方法去除滋养细胞，将得到的内细胞团（ICM）接种于丝裂霉素C灭活的小鼠胚胎成纤维细胞（MEFs）上培养5～8d，每4～7d传代1次，分别取不同代的hESC进行碱性磷酸酶（AKP）染色、转录因子OCT-4、阶段特异性胚胎抗原（SSEA）SSEA-4、SSEA-1、肿瘤排斥抗原（TRA）TRA-1-60、TAR-1-81、核型及体内外分化全能性鉴定。     

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.