人内皮抑素基因的克隆与表达及其抑瘤作用的实验研究

目的 克隆人内皮抑素基因,检测其表达蛋白的生物学活性,应用该蛋白治疗裸鼠皮下人脑胶质瘤。方法 从人肝组织提取mRNA,用反转录聚合酶链反应技术克隆人内皮抑素基因,将其重组人pUC19,双氰末端终止法测定其核苷酸序列。构建非融合表达载体pDH-endo,使其在DH5α内经温度诱导表达,纯化该表达蛋白并用鸡胚绒毛膜尿囊膜实验和内皮细胞抑制实验检测其活性。经荷瘤裸鼠皮下注射该蛋白治疗人脑胶质瘤。结果 成功获得551bp的人内皮抑素基因,测序正确。该诱导表达蛋白分子量为20000,具有抑制血管生成活性。该蛋白每天5、10或20mg/kg裸鼠皮下注射可抑制荷瘤裸鼠皮下人脑胶质瘤血管生成和肿瘤生长（抑瘤率分别为34．5％,76．1％和80．2％）。结论 人内皮抑素基因和表达和初步应用,为采用抗血管生成方法治疗脑胶质瘤等实体瘤研究奠定了实验基础。     

小鼠endostatin基因的克隆、表达及表达产物的活性鉴定

目的 克隆并表达小鼠endostatin基因,并初步检测其血管生长抑制功能,为胶质瘤的基因治疗探索新的途径。方法 从小鼠肝组织用RT-PCR法扩增endostatin基因,重组人pUC19质粒,经测序证实无误后,重组人表达载体pDH,经温度诱导表达,并用鸡胚绒毛尿囊膜（CAM）实验检测表达产物活性。结果 从小鼠肝组织中成功扩增到endo-statin基因,测序与报道序列一致,重组进pDH后经温度诱导表达,在SDS-PAGE电泳上得到一条新的蛋白表达带。分子质量为20ku,纯化后的蛋白能明显抑制CAM的血管生长。结论 成功构建了小鼠endostatincDNA的重组克隆pDH-Endo,并在大肠杆菌中获得高效表达。在CAM实验中表明纯化后的蛋白具有明显血管抑制作用。为利用endostatin进行脑胶质瘤等实验 全瘤的基因治疗奠定了实验基础。     

小鼠endostatin基因的克隆与C6鼠脑胶质瘤的治疗

目的 克隆小鼠ｅｎｄｏｓｔａｔｉｎ基因,检测其表达蛋白的生物学活性,应用该蛋白治疗大鼠Ｃ６脑胶质瘤。方法 从小鼠胶原ＸⅧｃＤＮＡ用ＰＣＲ法克 ｅｎｄｏｓｔａｔｉｎ基因,重组入ＰＵＣ１９,测序后构建非融合表达载体ｐＤＨ－ｅｎｄｏｓｔａｔｉｎ,使其在ＤＨ５α内经温度诱导表达,纯化该表达蛋白并用鸡胚绒 囊膜 内皮细胞抑制实验检测其活性,经荷瘤大鼠皮下注射该蛋白治疗其脑胶质瘤,结果 成功获得５５１ｂｐ的ｅ     

Endostatin基因真核表达载体的构建及活性鉴定

目的：构建endostatin基因的真核表达载体,并将其转染C6脑胶质瘤 细胞,探讨其体外抑制血管内皮细胞生长的活性。方法：利用PCR将大鼠血清蛋白分泌信号连接在endostatin基因的碳末端,构建成真核表达载体pcDNA3-Sendostatin,利用Lipofectamine法将其转染C6脑胶质瘤细胞,采用细胞增殖实验进行endostatin表达蛋白体外活性鉴定。结果：成功构建了endostatin基因真核表达载体,转染该载体的C6脑胶质瘤细胞可分泌表达抑制血管内皮细胞增生的endostatin蛋白。结论：endostatin是一种有效的血管生成抑制剂,可进一步用于在体肿瘤的抗血管生成治疗。     

原核表达的血管抑素K (1-3)蛋白对脑胶质瘤的生长抑制作用

目的获得具有抗血管生成活性的 angiostatin K(1-3)[AK(1-3)]基因的原核表达蛋白,应用该蛋白治疗裸鼠皮下人脑胶质瘤.方法构建和鉴定 pDH-AK(1-3)载体并在大肠杆菌DH5α内表达,诱导后用 SDS-PAGE分析、经亲和层析纯化后行Western Blot鉴定,以人脐带静脉血管内皮细胞抑制实验和鸡胚绒毛膜尿囊膜实验验证其生物学活性,应用AK (1-3) 蛋白治疗裸鼠皮下人脑胶质瘤.结果获得抑制人脐带静脉血管内皮细胞生长活性的AK(1-3) 原核表达蛋白(Mr为35×103),其蛋白总量占菌体总蛋白的12 %,该纯化表达蛋白5 mg·kg-1·次-1和10 mg·kg-1·次-1组均可抑制裸鼠皮下人脑胶质瘤生长,抑瘤率分别为34.5%和64.6%.结论该实验为应用抗血管生成药物治疗人脑胶质瘤等实体瘤研究奠定了初步基础.     

Endostatin裸DNA对人脑胶质瘤的抑瘤效应

目的：探讨endostatin裸DNA对裸鼠皮下脑胶质瘤生长的影响特点。方法：用脂质体包埋法将endostatin基因的真核表达载体pcDNA-S-Endo注入裸鼠皮下SHG44人脑胶质瘤内,观察肿瘤生长情况并计算抑瘤率,结合免疫组化、电镜、微血管和瘤体坏死区计数,确定endostatin裸DNA治疗人脑胶质瘤的特点。结果：Endostatin裸DNA在荷瘤裸鼠体内获得表达,裸鼠皮下胶质瘤血管生成能力显下降,肿瘤生长受到抑制,抑瘤率达到41．6％。结论：Endostatin裸DNA能够抑制人脑胶质瘤生长41．6％。结论：Endostatin裸DNA能够抑制人脑胶质瘤生长,它是通过抑制肿瘤血管生成,导致肿瘤坏死实现的。本研究为应用裸DNA药物治疗人脑质瘤奠定了初步基础。     

小鼠内皮抑素基因的克隆及序列测定

目的 获取小鼠内皮抑素（endostatin)基因并进行序列测定,为今后研究其机制及应用创造条件。方法 以小鼠肝脏总RNA为模板,用RT-PCR法从中扩增endostatin基因,并将所得基因重组入pUC19载体质粒,采用自动测定仪及荧光素标记引物,测定目的基因序列。结果 从小鼠肝组织中成功扩增endostatin基因,莘成功克隆入pUC19载体。经酶切鉴定正确后,命名为pUC-Endo。经测序证实所获基因序列与已知的endostatin基因序列一致。结论 成功构建小鼠endo-statin基因的重组克隆pUC-Endo,为今后利用endostatin进行胶质瘤的抗血管生成治疗研究奠定了实验基础。     

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…     

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.     

Dr.Zhang Ren's Experience in the Acupuncture Treatment of Different Diseases with the Same Therapeutic Principle

Dr.Zhang Ren,the chief physician,is the chairman of Shanghai Acupuncture and Moxibustion Association.Having been engaged in medicine for about 40 years,he is experienced in treating various intractable diseases.In his long years of clinical practice,he advocates taking the TCM differentiation as the basis to seek for the acupuncture method for treatment of modern intractable diseases.The author of this essay had the fortune to follow Dr.Zhang in study.The following is a summary of Dr.Zhang's experience in the acupuncture treatment for different intractable diseases with the same therapeutic principle.