Cx43基因剔除小鼠心脏锥干部的异常发育

目的探讨Cx43基因缺陷小鼠心脏锥干部发育异常。方法选用胚胎（embryonic day,E）11．5d至出生后1dC57／BL6小鼠作为研究对象,根据基因型分为Cx43基因剔除纯合子（Cx43-／-）、杂合子（Cx43＋／-）及野生型（Cx43＋／＋）,采用聚合酶链反应方法鉴定基因型。免疫组化法测定横纹肌肌动蛋白α-SCA、平滑肌肌动蛋白α-SMA、神经嵴细胞的标志物AP-2α的表达;原位杂交方法检测AP-2α mRNA的表达。结果Cx43-/-出生后24h内即死亡。大体解剖见明显的右室流出道圆锥部异常膨隆。HE染色示右心室流出道壁大量异常小梁状组织增生突起,右室流出道腔明显狭窄。Cx43＋／-无明显异常。Cx43-／-近端流出道隔中央区域α-SCA的表达明显滞后。Cx43＋／-与Cx43-／-小鼠右室流出道与Cx43＋／＋比较可见比较强的α-SMA的表达,主要位于右侧锥干部的异常增生部位。Cx43-/-小鼠在E13．5流出道隔AP-2α蛋白及其mRNA水平表达均增多,且表达位置异常。结论Cx43 KO小鼠以右室流出道异常增生引起的梗阻性畸形为主要特征。Cx43 KO小鼠胚胎期近端流出道隔心肌化迟滞。Cx43 KO小鼠锥干部α-SMA的表达不能正常消退,心肌细胞发育不成熟。神经嵴细胞的发育异常可能参与了Cx43基因缺陷小鼠锥干部畸形的发病机制。     

连接蛋白40、45在连接蛋白43基因敲除小鼠胚胎心脏的时空表达

目的 检测连接蛋白(Cx)40、45在Cx43基因敲除小鼠胚胎心脏的时空表达.方法 选取Cx43基因敲除胎鼠,通过PCR方法鉴定其基因型,以Cx43基因敲除胎鼠纯合子作为研究对象,野生型为对照组,获取胎龄10.5、11.5、12.5、13.5、14.5、15.5 d纯合子及野生型各2只胎鼠,免疫组化检测Cx40、Cx45在心脏各部位的表达,SCIM显微图像分析系统对染色强度进行定量分析.结果 (1)野生型小鼠心室原始小梁网在胎龄10.5 d就已经有Cx40表达(A值为8.6),随后在心房、心室、小梁网、房室间隔和主、肺动脉壁表达逐日增加,胎龄14.5 d在心室部位达到高峰(A值为94.8),然后逐渐减少;Cx43基因敲除小鼠胚胎心脏发育中Cx40在各部位的时空表达趋势与野生型相似,胎龄10.5 d在肌小梁A值为7.9,胎龄14.5 d心室部位为75.8.纯合基因敲除型胎鼠Cx40的表达量明显弱于野生型.(2)野生型小鼠肌小梁部在胎龄10.5 d出现Cx45的表达(A值为20.0),随后在房室各部位相继表达,胎龄12.5 d在心房部位达到高峰(A值为49.6),然后表达逐渐减少;Cx43基因敲除小鼠胚胎心脏发育中Cx45在各部位的时空表达趋势与野生型相似,胎龄10.5 d在肌小梁A值为17.8,胎龄12.5 d心房部位为31.5.纯合基因敲除型胎鼠Cx45的表达量明显弱于野生型.结论 Cx40和Cx45在Cx43基因敲除小鼠纯合子胎龄10.5～15.5 d这一心脏分隔、瓣膜发育的关键时期表达异常,可能与Cx43基因敲除小鼠心脏的异常发育有关.     

连接蛋白43基因敲除小鼠胚胎心脏流出道组织中转录因子的表达及意义

目的 研究连接蛋白43（Cx43）基因敲除小鼠胚胎心脏近端流出道组织中转录因子的改变,从分子水平探讨Cx43基因敲除小鼠胚胎心脏近端流出道发育异常的原因。方法 以胎龄13．5d和14．5d的Cx43基因敲除、Cx43杂合子和Cx43野生型鼠胚心脏近端流出道部分为研究对象。分别提取总RNA,反转录成cDNA;并在体外转录为cRNA,同时进行生物素标记及片段化;再与Affymetrix 4302．0小鼠全基因组芯片进行杂交。杂交信号经扫描后,应用相关生物信息软件分析基因表达情况。用实时定量逆转录（RT）PCR的方法对基因芯片筛选出的与心脏发育相关的部分转录因子进行验证。结果 与Cx43野生型组相比,Cx43基因敲除组表达差异的基因中,有6个是与心脏发育相关的转录因子。实时定量RT—PCR验证了其中3个基因：Soxll、Foxpl和Tbx20。在胎龄13．5d,Cx43基因敲除鼠胚Sox11、Foxp1的表达量均明显低于Cx43野生型组（4．76±0．19 vs 5．34±0．25,5．08±0．28 vs 5．64±0．15,均P〈0．01）;Tbx20在各组间差异不明显。胎龄14．5d,各基因Sox11、Foxp1以及Tbx20的表达量在基因敲除组均明显低于Cx43野生型组（4．71±0．27 vs 5．00±0．19,5．25±0．31 vs 5．77±0．16,7．05±0．17 vs 7．43±0．25,均P〈0．05）。其变化趋势与基因芯片结果基本一致。结论 心脏特异性的转录因子Foxp1、Sox11和Tbx20等的表达异常可能与Cx43基因敲除小鼠流出道的异常发育有关。     

Connexin43基因缺陷新生小鼠心脏组织病理学研究

目的观察不同程度Connefin43基因缺陷对新生小鼠心脏发育的影响。方法采用Cx43基因敲除(Cx43 KO)和CMV43^CT两种Cx43基因缺陷的小鼠模型,聚合酶链反应鉴定基因型。然后将新生小鼠心脏分离固定,HE染色观察心脏的形态结构。普通C57BL6／SJ小鼠作为对照。结果所有11只纯合型Cx43 KO小鼠生后1d内均死亡,心脏表现为严重右室流出道梗阻。在20只纯合型CMV43^CT新生小鼠中,有12只生后2d内死亡,其心脏不仅表现有右室流出道梗阻,还出现了房间隔缺损、室间隔缺损等其他心脏畸形;而另外8只未见心脏异常。所有杂合型Cx43 KO和CMV43^CT基因缺陷的小鼠生后均存活,心脏病理学检测未见异常。结论Cx43基因缺陷与心脏发育异常有密切的关系,但不同类型和不同程度的Cx43基因功能缺失对心脏发育的影响也不尽相同。     

应用PCR测定Cx43转基因小鼠的基因型

目的：应用聚合酶链反应(PCR)技术，检测Cx43(connexin 43)转基因小鼠的基因型。方法：采用CMV43转基因小鼠和Cx43剔除小鼠分别与野生型小鼠杂交繁殖，切取子代小鼠尾巴制备DNA，应用PCR技术扩增DNA，观察目的基因的阳性率。结果：CMV43转基因小鼠子代66个，dhfr PCR检测阳性率为31．8％。Cx43基因剔除小鼠子代21个，分别与Cx43野生型引物和Cx43剔除引物进行PCR，两者均阳性( ／ )12个，百分率为57．1％，前者阳性而后者阴性(杂合子 ／-)7个，百分率为33．3％，两者均阴性(纯合子-／-)2个，百分率为9．6％。野生型小鼠经Cx43野生型引物PCR检测结果均为阳性。结论：PCR技术用于Cx43转基因小鼠基因型的检测敏感度高，特异性强，易操作，可用作Cx43转基因小鼠心血管畸形模型研究的初选方法。     

右肺动脉异常起源于主动脉的诊断治疗分析

目的 评价右肺动脉异常起源于主动脉(AORPA)的诊断、治疗方法及效果.方法 回顾性分析1992年5月至2007年7月北京安贞医院14例AORPA患者术前诊断及手术治疗疗效资料.12例患者分别采用右全肺切除术、右肺动脉经人工血管或同种主动脉与主肺动脉吻合术、右肺动脉与主肺动脉直接吻合术、双片法右肺动脉与主肺动脉吻合术,同期修补合并畸形.结果 8例单纯超声心动图检查者中漏诊2例,6例右心室及主动脉造影和多排CTA/MRI检查者中无漏诊.2例因艾森曼格综合征未行手术,12例手术者围术期死亡2例.术后随访(5.15±3.48)年,晚期死亡1例,生存9例患者超声心动图复查左心室射血分数43%～52%,2例右肺动脉吻合口狭窄.心功能Ⅰ级5例,Ⅱ级4例.结论 右心室及主动脉造影或多排CTA/MRI检查可减少AORPA漏诊室.手术治疗AORPA的早期及中长期效果较好,但应尽早手术以提高疗效.自体组织加宽或加长右肺动脉进行右肺动脉重建术式吻合口狭窄的概率优于应用非自体组织重建右肺动脉术式.     

Cx43在神经管畸形发生中神经上皮的表达及意义

目的观察缝隙连接蛋白43(Connexin43, Cx43)在神经管畸形(neural tube defects, NTDs)发生中神经上皮的表达,以探讨Cx43与NTDs发生的关系。方法用过量RA建立小鼠胚胎NTDs模型。用SABC免疫组化方法和图像处理系统,分析比较实验组和对照组神经管发育情况以及Cx43蛋白表达的位置和强度。结果对照组神经管发育良好并适时闭合,实验组神经管发育不良而且不能闭合,脑泡不能发育;Cx43蛋白表达于神经上皮细胞的细胞膜和细胞质内,而且实验组明显高于对照组（P＜0.05）;Cx43蛋白的表达表现出时间的差异性。结论Cx43蛋白定位于神经管上皮细胞的胞膜和胞质内,且有时间差异性。Cx43蛋白的过高表达可能与RA致NTDs密切相关。     

Cx43基因启动子甲基化下调乳腺癌Cx43的表达

目的：探讨乳腺癌细胞中Cx43蛋白的表达与基因启动子甲基化的关系。方法：免疫组化检测乳腺标本中Cx43蛋白的表达,甲基化PCR检测Cx43启动子甲基化的频度。5-氮杂脱氧胞苷培养乳腺癌MD-MBA-231细胞后,RT-PCR乳腺癌细胞中Cx43 mRNA,免疫组化、Western Blot检测Cx43蛋白表达,甲基化PCR检测启动子甲基化的变化。结果：与正常乳腺组织相比,乳腺癌中Cx43蛋白的表达下调,启动子甲基化频度高。在应用5-Aza-dC处理前,MD-MBA-231 Cx43基因呈甲基化状态,经4.0μmol/L的5-Aza-dC作用48h后,乳腺癌细胞Cx43基因甲基化逆转,Cx43基因mRNA及蛋白表达增加。结论：Cx43在乳腺癌组织中表达下调可能与启动子甲基化有关。5-Aza-dC能逆转乳腺癌细胞MD-MBA-231的Cx43基因异常甲基化,促进Cx43基因的再表达。     

双腔右心室的超声心动图诊断

目的：探讨双腔右心室(DCRV)的超声心动图特点及误诊原因。方法：总结自1994年5月至2002年6月问经手术证实为DCRV患者13例的病例资料，分析DCRV超声诊断的特点，并与手术结果对比。结果：通过不同切面显示右心室腔内室上嵴或嵴下水平至右室游离壁异常肌束横跨；右室肥厚呈楔形向右室腔凸起，室间隔呈舌形凸向右室腔，两者相对，中间形成狭窄口，彩色多普勒血流显像示经狭窄处的五彩镶嵌血流信号，连续多普勒检测跨狭窄处压力阶差可反映梗阻的程度；超声心动图检查结果与手术相符者有11例，误诊2例，符合率为84．6％。结论：超声心动图在诊断DCRV及合并畸形等方面具有可靠价值。     

EPS对肺动脉高压大鼠右心室重构的影响及机制

探讨依帕司他(EPS)对肺动脉高压大鼠右心室重构的影响及机制。采用野百合碱(MCT)诱导的方法建立肺动脉高压大鼠模型。分别给予为50 mg/kg和100 mg/kg EPS连续干预4周,观察大鼠血流动力学指标、右心室重构及其心肌形态和心肌Cx43表达。结果显示,EPS能够降低右心室收缩压(RVSP)、平均肺动脉压(mPAP)、肺动脉收缩压(PASP)、右心室肥厚指数(RVHI),右心室质量指数(RVMI)及增加心肌Cx43表达,并呈剂量依赖效应。结果说明,EPS能改善MCT诱导的肺动脉高压大鼠右心室重构,机制可能与其增加心肌Cx43表达有关。     

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.