p16蛋白及其基因在原发性肝细胞癌中的表达缺失性研究

目的通过p16蛋白及其基因在原发性肝细胞癌中的表达缺失研究,探讨p16基因与肝癌的关系.方法收集我院原发性肝癌32例,用免疫组织化学SP法、聚合酶链式反应(PCR)方法分别检测肝癌及癌旁组织中p16蛋白的表达及其基因的缺失情况.结果32例肝癌中,p16蛋白阴性表达率为62.5%(20/32),癌旁组织中p16蛋白阴性表达率为3.13%(1/32),两者差异有显著意义(P＜0.01).p16蛋白的表达与肝癌分化程度关系密切(P＜0.05),而与癌灶体积、甲胎球蛋白(AFP)值的差异无显著意义(P＞0.05).p16基因在癌组织中的缺失率25%(8/32),在癌旁组织为3.13%(1/32),相差显著(P＜0.05).p16基因缺失与肝癌分化程度、癌灶体积、AFP值无关(P＞0.05).结论p16基因缺失与肝癌发生关系密切,提示p16蛋白可能是预测病人预后的一个指标,但仍需进一步确证.     

PTEN和p53在肝细胞癌中的表达及其相关性

目的：探讨肝细胞癌（hepatocellular carcinoma,HCC）第10号染色体缺失的磷酸酶张力蛋白同源物基因（gene of phosphate and tension homology deleted on chromsome ten,PTEN）和p53蛋白的表达及其临床意义.方法：采用免疫组织化学EliVision方法测定70例HCC组织和40例相应的癌旁组织中PTEN和p53蛋白的表达,分析两个指标与临床病理特征之间的关系以及它们之间的相关性.结果：PTEN在HCC组织中的阳性率为57.1%,显著低于癌旁组织的95.0%（P＜0.01）.在肿瘤分期晚和有淋巴结转移的患者中,PTEN表达均降低（P〈0.01）.p53蛋白在HCC组织中表达率为60.0%,显著高于癌旁组织的15.0%（P＜0.01）.在肿瘤大小、病理分级、TNM分期和淋巴结转移等临床病理指标上,p53蛋白表达差异均无统计学意义（P〉0.05）.PTEN与p53蛋白表达在HCC组织中呈负相关关系（P＜0.05）.结论：PTEN低表达与HCC进展有关,有可能作为评价肝癌预后的指标.     

蛋白翻译起始因子C2在原发性肝细胞肝癌中表达的意义

目的：研究C2 mRNA及其蛋白在原发性肝细胞肝癌（HCC）中的表达及意义。方法：用原位杂交检测C2 mRNA在21例HCC及其癌旁组织中的表达,ABC免疫组化法检测C2蛋白60例HCC及其42例癌旁组织中的表达,Western blot检测C2蛋白在HCC及其癌旁组织中的表达。结果：21例HCC及其癌旁组织中,C2 mRNA阳性率分别占23.8%（5/21）和85.7%(18/21)。60例HCC及42例癌旁组织中,C2蛋白阳性分别占27.3%(17/60)和83.3%（35/42）。27例肝硬变中,C2蛋白阳性率为77.8%(21/27)。χ^2检验：C2 mRNA及其蛋白在HCC癌旁组织中表达明显高于癌组织（P＜0.001);C2蛋白在肝硬变中的表达明显高于HCC癌组织（P＜0.001);C2的表达与患年龄、HBsAg及AFP有明显关系（P＜0.001);碉与癌组织的分化程度、肿瘤大小、淋巴转移无并;Western bolt与免疫组化结果一致。结论：C2基因表达下调可能与HCC的发生、发展及早期诊断。     

原发性肝细胞癌中p16和cyclinD1蛋白表达及p16突变研究

目的：研究p16和cyclinD1蛋白表达及p16基因第2外显子突变与原发性肝细胞癌（HCC）发生的关系。方法：利用SP免疫组织化学方法与多聚酶链反应－单链构象多态性（PCR－SSCP）分析技术分别检测44例HCC及癌旁肝组织p16和cyclinD1蛋白表达和12例新鲜手术肝癌组织的p16基因突变。结果：HCC中p16蛋白阳性率和阳性信号强度明显低于癌旁肝组织（P＜0．01）,而cyclinD1蛋白阳性率和阳性信号强度明显高于癌旁肝组织,12例新鲜HCC标本中p16基因第2 子突变率为16．7％（2／12）,癌旁组织未发现p16基因第2外显子突变;有p16基因突变的HCCp16蛋白表达呈阴性。结论：P16蛋白失活或／和缺乏可能是肝细胞增殖失控的重要因素之一;HCC存在P16基因第2外显子突变,但不是频发事件;在HCC形成过程中,P16基因异常与cyclinD1过度表达可能存在协同作用。     

原发性胰腺癌中p16基因突变及其蛋白表达的研究

目的：探讨p16基因与胰腺癌发生的关系。方法：采用PCR基础缺失分析，PCR－SSCP及免疫组织化学方法检测28例胰腺癌组织，相应癌旁组织和4例正常胰腺组织中p16基因缺失，突变及其蛋白表达状况，并结合临床病理资料进行分析，结果：28例原发性胰腺癌组织中4例发生p16基因纯合缺失。5例发生p16基因突变，p16基因变异频率为32．1％，28例癌旁组织和4例正常胰腺组织中未发现纯合缺失及突变。p16基因变异频率与淋巴结转移（P＜0．025），转移淋巴结个数（P＜0．01）及临床分期（P＜0．05）密切相关。28例原发性胰腺癌的癌组织，癌旁组织和4例正常胰腺组织p16蛋白阳性表达率分别为57．1％（16／28），85．7％（24．28）和100％（4／4），癌组织中p16蛋白阳性表达率与组织学分化（P＜0．05），淋巴结转移（P＜0．05），转移淋巴结个数（P＜0．05）及临床病理分期（P＜0．05）密切相关。结论：p16基因变异及其蛋白失表达可能是胰腺癌发生发展过程中重要因素之一，而且二者均可作为评估胰腺癌恶性程度及其侵袭能力的有用指标，存在p16基因变异及蛋白失表达的原发性胰腺癌具有更强的恶性潜能。     

p16蛋白及其基因在原发性肝细胞癌中的表达缺失及意义

目的:通过p16蛋白及其基因在原发性肝细胞癌中的表达缺失研究,探讨p16基因与肝癌的关系.方法:收集我院原发性肝癌32例,用免疫组织化学SP法、聚合酶链式反应(PCR)方法分别检测肝癌及癌旁组织中p16蛋白的表达及其基因的缺失情况.结果:32例肝癌中,p16蛋白表达缺失率为2.5%(20/32),癌旁组织中p16蛋白表达缺失率为3.13%(1/32),两者差异有显著意义(P<0.01).P16蛋白的表达与肝癌分化程度关系密切(P<0.01).P16基因在癌组织中的缺失率25%(8/32,均为p16蛋白表达缺失者),在癌旁组织为3.13%(1/32),相差显著(P<0.05).结论:p16基因缺失与肝癌发生关系密切,提示p16蛋白可能是预测患者预后的一个指标,但仍需进一步确证.     

原发性肝细胞癌P53、P16和c-erbB-2蛋白表达意义

目的：研究P53、P16和c-erbB-2蛋白表达与原发性肝细胞癌发生物学行为的关系。方法：应用免疫组化链霉素抗生素蛋白－过氧化物酶连结法（LSAB）检测60例原发性肝细胞癌和癌旁肝组织P53、P16和c-erbB-2蛋白表达。结果：P53、P16和c-erbB-2在原发性肝细胞癌中的阳性率分别为41．6％、35％和58．33％，癌旁肝组织的阳性率分别为8．33％、88．33％和21．67％，差异有显著性（P＜0．01或P＜0．05）。P53和c-erbB-2与肝细胞癌的病理组织学分级具有一定程度的相关性，P16蛋白与病理组织学分级有显著差异（P＜0．05）。P16、c-erbB-2和P53均与转移有关。P53蛋白表达与c-erbB-2蛋白表达具有显著的协同性（P＜0．05），与P16蛋白表达呈负相关性（P＜．05）。结论：肿瘤的多因素分析比单因素分析更有价值，癌基因c-erbB-2和抑癌基因P53、P16蛋白的表达异常及协调作用在原发性肝癌的发生、发展中起重要作用。     

肝细胞癌及癌旁肝组织中PTEN表达与MAPK磷酸化的相关性

目的：研究肝细胞癌（HCC）及癌旁肝组织中PTEN表达与丝裂素激活的蛋白激酶（mitogen-activated protein kinase,MAPK)磷酸化的相关性。方法：利用SP免疫组织化学检测75例HCC及其癌旁肝组织中PTEN蛋白和p42/44^MAPK的表达。结果：①HCC中PTEN蛋白的阳性率（62．7％）和阳性强度明显低于癌旁肝组织（89．3％）（P＜0．01），且阳性信号强度与HCC分化程度相关（P＜0．01），即HCC分化愈差，PTEN蛋白表达愈弱。②HCC中p42/44^MAPK的阳性率（85．3％）和表达强度明显高于癌旁肝组织（34．7％）（P＜0．01），但阳性信号强度与癌细胞分化程度无关（P＞0．05）。③相关性分析表明，HCC及癌旁肝组织中PTEN蛋白表达强度和MAPK磷酸化程度呈明显负相关（P＜0．01）。结论：提示在HCC发生中，可能是由于PTEN蛋白表达降低或缺失，不能有效抑制由致癌因子异常激活的Ras/Raf/MAPK信号转导通路，从而使肝细胞异常增殖和发生恶性转化。     

高表达的eEF-2K基因沉默对肝细胞癌增殖、侵袭和凋亡的影响

目的：检测eEF-2K基因在肝细胞癌（HCC）组织内的表达并分析其表达水平对HCC细胞增殖、侵袭和凋亡的影响。方法：用RT-qPCR及Western blot方法检测了64例肝癌组织及癌旁肝组织、五种HCC细胞系LM3, Hep3B, Huh7, 97H, HepG2及人正常肝细胞系 LO2中eEF-2K基因在mRNA及蛋白水平的表达。用eEF-2K 特异性siRNA沉默LM3及Hep3B细胞性中eEF-2K基因后，经qPCR检测确认。随后用CCK-8，Transwell方法与流式细胞技术检测eEF-2K基因沉默对增殖、侵袭与凋亡的影响。结果：相较癌旁组织，HCC组织内eEF-2K mRNA与蛋白表达显著上调（P＜0.05）；与正常人肝细胞系LO2相比，肝癌细胞系LM3, Hep3B, 97H, HepG2, Huh7中的eEF-2K mRNA和蛋白表达分别升高（Ｐ＜0.05）。在LM3及Hep3B细胞系中用特异性siRNA沉默eEF-2K后，与非特异性siRNA对照组相比，细胞增殖活性和侵袭能力显著降低（P＜0.05），凋亡率显著升高（P＜0.05）。TCGA数据库分析结果表明eEF-2K高表达与肝癌预后不良相关（P＜0.05）。结论：eEF-2K蛋白在肝癌组织和细胞中高表达且与预后不良相关；沉默eEF-2K后肝癌细胞增殖和侵袭能力降低并诱发凋亡，提示eEF-2K可用作HCC潜在的预后标志物和治疗靶点。     

Wnt-5a在原发性肝细胞癌发生中的表达及临床意义

目的 检测Wnt-5a基因和基因产物在原发性肝细胞癌中的表达,探讨Wnt-5a基因在肝癌发生过程中的作用和临床意义.方法 采用实时定量PCR技术,检测Wnt-5a mRNA在26例新鲜肝细胞癌和癌旁组织中的含量;用免疫组化法观察Wnt-5a蛋白在85例肝细胞癌和癌旁组织、15例肝硬化组织中的表达.结果 Wnt-5a mRNA在肝癌及癌旁组织中的相对含量分别为0.102 127±0.158 620和0.020 106±0.022 075,两者间差异有统计学意义（P＜0.05）;Wnt-5a蛋白在肝细胞癌、癌旁组织和肝硬化中的阳性率分别为21.2％ （18/85）、81.26％ （69/85）和86.7％ （13/15）,Wnt-5a在肝癌组织中的阳性表达显著低于肝硬化和癌旁组织,差异有统计学意义（P ＜0.01）;Wnt-5a蛋白在肝癌中的表达与肿瘤的TNM分期和血清甲胎蛋白（AFP）含量有关（P＜0.05）.结论 Wnt-5a基因在HCC基因转录中呈上调表达,而在蛋白水平显示为缺失和下调表达,提示肿瘤细胞是在蛋白翻译水平而不是转录水平干扰了Wnt-5a基因功能.     

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.