西洛他唑改善脑梗死急性期小鼠脑血流量作用机制研究

目的观察西洛他唑（CLZ）改善脑梗死急性期小鼠脑血流量的效果并探讨其作用机制。方法12只8周龄雄性Icr小鼠建立左侧大脑中动脉永久闭塞模型,分为CLZ组、PBS组及HPβCD组,每组4只,给予CLZ悬浊液、PBS溶液及HPβCD溶液后于不同时间点进行ROI区域（L）及对侧映射区域（R）的脑血流量测定。另将CLZ、PBS和HPβCD分别加入培养液中对小鼠大脑由来血管内皮细胞进行培养,6d后采用Westernblot法检测细胞磷酸化内皮型一氧化氮合成酶（p-eNOS）及内皮型一氧化氮合成酶含量（eNOS）并对比分析。结果CLZ组小鼠注射药物后半暗带区域脑血流量较其他两组小鼠有所增加;注射后20、30及60min时的L/R比值均高于0min,且注射后20、30min时的L/R比值均高于PBS组、HPβCD组,差异均有统计学意义（均P＜0.05）。结论CLZ可改善脑梗死急性期梗死区域脑血流量,其作用机制可能为促进p-eNOS活性的表达。     

芹菜素通过VEGF通路对大脑中动脉阻塞后大鼠脑缺血半暗带血管神经再生的影响

目的：探讨芹菜素促大脑中动脉阻塞（MCAO）后大鼠脑缺血半暗带血管生成和神经再生的作用,以及是否与血管内皮生长因子（VEGF）信号通路有关。方法：将144 只SD大鼠分为假手术组（S7、S14）、模型组（M7、M14）、芹菜素模型组（AM7、AM14）和芹菜素+VEGF受体（VEGF-R）抑制剂模型组（AIM7、AIM14）。构建MCAO模型后,采用改良神经行为学评分（mNSS）和TTC染色评估神经功能和脑梗死体积,采用Western blot、免疫组化和免疫荧光评估脑缺血半暗带中VEGF表达水平以及血管生成和新生神经元的变化。结果：与S组比较,M组、AM组和AIM组均出现不同程度的神经行为学异常和白色梗死灶,AM组较同期M组的mNSS评分降低、脑梗死体积减小（P ＜0.05）;缺血半暗带AM组与同期M组比较VEGF表达增加（P ＜0.01）,AM7 组较M7 组的脑微血管密度增高（P ＜0.05）;AM7组BrdU/Nestin和AM14组BrdU/NeuN的双标阳性细胞数目均较同期M组明显升高（P ＜0.01）;但VEGF-R抑制剂均可下调芹菜素的上述作用。结论：芹菜素可通过上调VEGF信号通路促进脑缺血半暗带血管生成和神经再生,改善局灶性脑缺血再灌注所致的神经功能缺损。     

静脉注射骨髓基质细胞对脑梗死大鼠血管内皮生长因子表达的研究

目的：观察静脉注射的骨髓基质细胞（MSC）在大脑中动脉闭塞（MCAO）再灌注模型大鼠脑内分布及血管内皮生长因子（VEGF）表达。方法：采用Longa线栓法建立Wistar大鼠MCAO再灌注模型；从大鼠后肢骨髓分离、培养MSc；CD34、CD44、CD54免疫细胞化学染色鉴定MSC；5-溴脱氧尿苷（BrdU）标记的MSC注射给动物模型，免疫组织化学染色观察BrdU阳性细胞分布及VEGF表达。结果：体外培养的MSC具有多态性和贴壁生长特性，多次传代后细胞生物学特性没有明显改变；MSC表达CD34（-）、CD44（＋）、CD54（＋）；治疗组BrdU阳性细胞主要分布于梗死灶周围。VEGF表达比对照组多（P〈0．05）。结论：静脉注射的MSC能游走、迁移至太鼠缺血脑组织并存活，同时促进VEGF表达。     

血管内皮祖细胞对共同植入缺血再灌注模型大鼠脑内的神经干细胞增殖、凋亡和血管形成的调节

目的研究血管内皮祖细胞(endothelial progenitor cells,EPCs)与神经干细胞(neuron stem cells,NSCs)构建的复合移植体对移植入缺血再灌注模型大鼠缺血半暗带的神经干细胞增殖、凋亡和血管重建的影响。方法线栓法建大鼠脑缺血再灌注模型;血管内皮祖细胞、层粘连蛋白和神经干细胞构建的移植复合体移入模型鼠脑缺血半暗带;免疫组化观察Brdu标记的移入脑内神经干细胞和缺血半暗带血管新生,免疫荧光双标观察神经干细胞的凋亡情况。结果移植后各时间点Brdu标记的神经干细胞数NSCs+EPCs组明显高于NSCs组(P<0.05);NSCs+EPCs组在各时间点的凋亡率明显低于NSCs组(P<0.05);各时间点NSCs+EPCs组血管新生数量明显多于EPCs组、缺血对照组和NSCs组(P<0.05)。结论血管内皮祖细胞可以促进移植入缺血再灌注模型大鼠缺血半暗带的神经干细胞增殖,减少其凋亡;2种细胞共移植可以促进缺血半暗带的血管新生。     

大鼠局灶脑缺血后内皮前体细胞的相关实验研究

目的探讨缺血性卒中后外周血CD34^＋细胞和脑组织AC133抗原的变化。方法本研究采用大鼠大脑中动脉缺血再灌注线栓法模型,缺血2h。将成年SD雄性大鼠随机分为正常组、假手术组、动物模型组,分别取缺血再灌注1、3、6、12、24、48、72h、4、7、14d等作为观察点。取再灌注1、3、6、12、24、48、72h、4、7、14d外周血标本,采用流式细胞术检测外周血单个核CD34＋细胞平均荧光强度。采用免疫组化染色法检测2,3、4、7d脑组织AC133抗原表达。每个观察点5只大鼠,共60只大鼠,根据神经功能计分纳入实验。结果①大鼠脑缺血／再灌注3d外周血单个核CD34^＋细胞平均荧光强度明显降低,直到7d。14d恢复到基线水平。②AC133抗原在再灌注4d梗死半球半暗带区域的血管内皮细胞表达阳性,3、7d组无阳性表达。结论脑缺血再灌注后早期外周血CD34^＋干细胞明显下降,脑梗死半暗带区域部分血管内皮细胞出现AC33抗原的表达,提示内源性血管内皮干细胞可能参与脑缺血损伤的血管修复。     

骨髓基质细胞治疗大鼠局灶性脑缺血的疗效评价

目的：观察静脉注射骨髓基质细胞（BMSC）治疗大鼠局灶性脑缺血的疗效。方法：取健康Wistar大鼠45只,雌雄不限,随机分为骨髓基质细胞组（治疗组）,生理盐水组（实验对照组）和空白对照组。Fridenshtein方法分离培养骨髓基质细胞,免疫细胞化学法鉴定骨髓基质细胞,改良Longa线栓法制作大鼠大脑中动脉栓塞（MCAO）1h模型。栓塞24h后,治疗组股静脉注射经BrdU标记的终浓度为3×10^6个/ml骨髓基质细胞1ml,实验对照组股静脉注射1ml生理盐水,空白对照组股静脉不注射。采用免疫组化法检测脑内BrdU阳性细胞的表达,神经功能损伤评分（NSS）检测大鼠神经系统功能,进行疗效评价。结果：免疫细胞化学方法鉴定骨髓基质细胞为CD34（-）,CD54（＋）、CD106（＋）、CD44（＋）。栓塞后3,7,14d在缺血侧半球可以发现大量BrdU阳性细胞,对侧半球也可见少量BrdU阳性细胞。治疗组和两个对照组相比各时间点的神经功能评分均偏小,在栓塞14d时有显著性差异（P〈0.05）。结论：骨髓基质细胞（BMSC）静脉注射后大鼠的神经功能明显得到了恢复,故认为静脉注射骨髓基质细胞治疗局灶性脑缺血简便易行,并且有效。     

人源脐带间充质干细胞对脑缺血再灌注小鼠神经功能及神经炎症的影响

目的探究人源脐带间充质干细胞对脑缺血再灌注损伤小鼠神经功能及神经炎症的影响。方法将96只野生型C57BL/6小鼠随机分为假手术组、磷酸盐缓冲液(PBS)处理组、WJ-MSCs细胞移植组,每组32只。PBS处理组和细胞移植组小鼠通过线栓法构建大脑中动脉阻塞再灌注模型;术后24h,通过脑立体定位技术注射细胞悬液或PBS溶液。每组取6只小鼠,于细胞移植后对小鼠腹腔注射BrdU溶液,持续5d。于术前及术后1、6、12、24d对各组小鼠进行改良神经功能损伤评分;术后6d时,采用神经元标志物NeuN免疫荧光单标记染色技术检测缺血侧大脑梗死灶及梗死灶周围区皮质神经元的变性丢失情况;采用RT-qPCR技术检测各组小鼠缺血侧大脑半球炎症因子的表达量;采用小胶质细胞/巨噬细胞标志物Iba1及增殖标记物BrdU免疫荧光双标记染色技术检测小胶质细胞/巨噬细胞的活化和增殖情况。结果与PBS处理组相比,WJ-MSCs细胞移植组小鼠神经功能显著改善(P<0.01),且位于缺血侧大脑梗死灶及梗死灶周围区皮质的存活神经元数目显著增多(P<0.01)。与PBS处理组相比,WJ-MSCs细胞移植组小鼠缺血侧脑组织的抗炎因子表达量升高,促炎因子表达量下降,差异具有统计学意义(P<0.05)。与PBS处理组相比,WJ-MSCs细胞移植组小鼠缺血侧梗死灶和梗死灶周围区皮质小胶质细胞/巨噬细胞的活化显著抑制(P<0.05),且小胶质细胞/巨噬细胞的增殖明显降低(P<0.05)。结论人源脐带间充质干细胞可抑制神经炎症反应并改善脑缺血再灌注小鼠的神经功能。     

西洛他唑与阿司匹林对脑保护作用的实验研究

目的探讨西洛他唑在脑梗死急性期的脑保护作用和对出血的影响。方法 100只小鼠分为假手术组、对照组、西洛他唑组(10 mg/kg)、阿司匹林组(30 mg/kg)。建立小鼠大脑中动脉梗死3 h再灌注21 h模型,梗死后24 h进行脑含水量、脑梗死体积、脑出血量测定和神经功能学评分。结果与对照组相比,西洛他唑组脑出血量、脑含水量无显著增加,梗死面积显著减小,神经功能学评分显著改善。而阿司匹林组脑水肿、脑出血量显著增加,并未改善神经功能学评分(P<0.05)。结论脑梗死急性期应用西洛他唑可以减小脑水肿和梗死体积,改善神经功能且并不增加出血风险;而阿司匹林则无明显脑保护作用且增加出血风险。     

小鼠同种异体内皮祖细胞移植治疗急性肺损伤的相关研究

目的：观察外源性内皮祖细胞（endothelial progenitor cells,EPCs）移植对急性肺损伤（Au）小鼠受损肺组织的保护作用并探讨其机制。方法：选用转基因EGFP-BALB／c小鼠作为细胞移植供体,获得骨髓单核细胞,体外培养扩增获得足够数量的表达绿色荧光蛋白的EPCs（EGFP-EPCs）。选用野生型BALB／c小鼠18只,随机分为假手术组、AU组及EPC治疗组,每组6只。假手术组气管内滴注PBS,其后经尾静脉注射PBS;Au组气管内滴注脂多糖溶液,其后经尾静脉注射PBS;EPC治疗组气管内滴注LPS,其后经尾静脉注射EGFP-EPCs。各组小鼠术后3d处死,留取肺组织标本。荧光显微镜下观察各组肺组织中EGFP-EPCs表达情况,HE染色评价各组肺组织损伤程度并进行肺损伤指数评分,比较各组肺组织湿干重比,行肺组织CD34免疫组化染色并测定肺微血管密度（MVD）。结果：各组小鼠建模过程顺利,无围手术期死亡。EPC治疗组小鼠肺组织冰冻切片中可观察到均匀附着于血管内皮的绿色荧光表达,而假手术组及ALI组则未见明显的绿色荧光。HE切片显示：LPS可致严重的肺组织损伤,而EPC治疗组肺损伤程度较AU组明显减轻,肺损伤指数明显低于ALI对照组（P〈0．001）。肺组织湿干重比Au组明显高于假手术组（P〈0．001）,而EPC治疗组较ALI组明显降低（P〈0．05）。免疫组化染色显示：EPC治疗组肺组织切片中CD34的表达明显增加,MVD值均明显高于假手术组及ALI组（P〈0．001）。结论：对ALI小鼠行外源性EPC移植治疗,输入体内的EPC可定向迁移并黏附于受损的肺毛细血管内皮,直接参与肺毛细血管内皮修复及重建,维持肺毛细血管内皮完整性,减少炎症细胞向肺间质、肺泡内的浸润及对肺组织的炎性损伤。同时,外源性EPC治疗提高了EPC向受损肺组织归巢及受损肺组织内的血管新生。     

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.