急性β射线皮肤损伤创面愈合过程中相关凋亡基因的表达

目的 研究大鼠急性β射线皮肤损伤创面愈合过程中相关凋亡基因Bcl-2、Bax和P53表达的动态变化,探讨急性β射线皮肤损伤创面难以愈合的机制.方法 雌性SD清洁级大鼠54只,随机分为3组:照射组(n=24)为急性深Ⅱ度β射线皮肤损伤动物模型,烫伤组(n=24)为深Ⅱ度热力烫伤动物模型,对照组(n=6)为正常大鼠.取不同时期创面组织,采用原位末端标记(TUNEL)、免疫组织化学等方法检测不同时相点各组大鼠创面局部相关凋亡基因Bcl-2、Bax和P53的变化.结果 (1)TUNEL结果显示,照射组大鼠创面出现后0、1、2、3周的积分吸光度(IA)值明显高于烫伤组,差异有统计学意义(P＜0.05).(2)免疫组化检测结果显示,照射组创面出现后0、1、2、3周Bax、P53的IA值明显高于烫伤组,差异有统计学意义(P＜0.05);而烫伤组Bcl-2的IA值明显高于照射组,差异有统计学意义(P＜0.05).结论 急性β射线皮肤损伤创面细胞凋亡出现较早、频度较高、消失推迟,可能是导致创面难以愈合的重要原因之一.     

大鼠β射线皮肤损伤创面愈合过程中Tie-2 mRNA的表达

目的研究大鼠急性β射线皮肤损伤创面愈合过程中酪氨酸激酶免疫球蛋白和表皮生长因子同源受体-2(Tie-2)mRNA的表达情况。方法将54只雄性SD清洁级大鼠随机分为3组。照射组(=24),直线加速器产生射线(45Gy)单次照射大鼠臀部皮肤40 mm×40 mm,建立急性射线皮肤损伤模型;烧伤组(=24),将直径30mm铜柱于100℃沸水中放置15 min,取出后在无外界压力下置于大鼠臀部皮肤8 s,建立深Ⅱ°热力烧伤模型;对照组(=6),正常大鼠。取不同时期创面皮肤组织,采用原位逆转录聚合酶链反应(RT-PCR),检测不同时点各组大鼠创面组织中Tie-2mRNA的表达。结果 A组大鼠照射后第2天皮肤出现充血性红斑,2周后脱毛,4周后出现溃疡创面,5周后创面仍未愈合。B组大鼠烧伤后创面即刻出现水泡,3d后肉芽增生,3～4周后创面瘢痕愈合。大鼠正常皮肤内Tie-2mRNA为可疑阳性;A组大鼠创面Tie-2mRNA表达伤后1周略有升高,并持续低表达;B组Tie-2 mRNA明显上调,2周达峰,3周时仍高于A组(均<0.05)。结论急性射线皮肤损伤创面愈合过程中Tie-2的持续低表达,不能形成峰值,可能是急性射线皮肤损伤创面难愈合的机制之一。     

β射线放射损伤后胶原代谢的变化

目的　探讨 β射线放射损伤后胶原代谢的改变情况。 方法　采用直线加速器照射制作动物的β射线损伤模型 ,同时对NIH 3T3细胞进行β射线外照射 ,观察胶原总量及Ⅰ、Ⅲ型胶原含量的改变 ;观察胶原降解酶MMPs- 1活性变化 ;同时测定损伤后细胞因子TGF - β1、IL - 6的变化情况。结果　β射线损伤后胶原总量变化不大 ,而Ⅰ型胶原含量下降 ,Ⅲ型胶原含量上升 ;MMPs- 1活性上升 ;TGF - β1、IL - 6表达量增加。结论　胶原代谢的变化在放射性皮肤损伤中起关键作用 ,TGF - β1、IL - 6在损伤后胶原代谢的调控机制中起重要作用     

Ⅳ型胶原酶在糖尿病大鼠肺组织表达及PKC活性变化的研究

目的：探讨Ⅳ型胶原酶(MMP2及MMP9)在糖尿病大鼠肺组织表达的变化及蛋白激酶C(PKC)的作用。方法：STZ腹腔注射制作糖尿病大鼠模型，4周后观察肺组织的病理改变，免疫组化方法检测MMP2及MMP9在糖尿病大鼠肺组织表达的变化，采用改良的Takay法测定PKC活性，蛋白质免疫印迹分析(Western—blot)及免疫组化方法检测TGF—β1表达含量的变化。结果：DM大鼠4周后肺泡间隔及毛细血管壁增厚，肺间质胶原成分增多，肺组织PKC活性增强，TGF—β1表达增多，MMP2及MMP9表达下降。结论：在糖尿病大鼠肺组织高糖环境下，PKC被激活导致TGF—β1表达增高，MMP2、MMP9表达下降，引起细胞外基质(ECM)合成降解失调，可能参与了糖尿病肺部并发症的发生及发展。     

血管生成素1在大鼠急性射线皮肤损伤创面愈合过程中的表达

目的研究大鼠急性射线皮肤损伤创面愈合过程中血管生成素1（Ang-1）的表达情况。方法将雄性SD清洁级大鼠54只分为3组。照射组（=24）,直线加速器产生射线（45 Gy）单次照射大鼠臀部皮肤40 mm×40 mm,建立急性射线皮肤损伤模型;烧伤组（=24）,将直径30 mm铜柱于100℃沸水中放置15min,取出后在无外界压力下置于大鼠臀部皮肤8 s,建立深Ⅱ°热力烧伤模型;对照组（=6）,无特殊处理。取不同时期创面皮肤组织,采用免疫组织化学（SP法）检测不同时相点各组大鼠创面组织中Ang-1的表达。结果免疫组化显示,烧伤组在伤后1、2及3周Ang-1表达积分吸光度值均要大于照射组,差异均有统计学意义（〈0.05）。结论急性射线皮肤损伤创面愈合过程中Ang-1持续低表达,不能形成峰值,可能是其损伤创面难愈合的机制之一。     

β射线放射损伤后胶原代谢的变化

目的 探讨β射线放射损伤后胶原代谢的改变情况。方法 采用直线加速器照射制作动物的β射线损伤模型,同时对ＮＩＨ３Ｔ３细胞进行β射线外照射,观察胶原总量及Ⅰ、Ⅲ型胶原含量的改变;观察胶原降解酶ＭＭＰｓ－１活性变化;同时测定损伤后细胞因子ＴＧＦ－β１、ＩＬ－６的变化情况。结果 β射线损伤后胶原总量变化不大,而Ⅰ型胶原含量下降,Ⅲ型胶原含量上升;ＮＭＰｓ－１活性上升;ＴＧＦ－β１、ＩＬ－６表达量增加。结论     

老龄大鼠肺内基质金属蛋白酶2、9及基质金属蛋白酶组织抑制因子1、2、3表达变化

目的:对比观察老龄大鼠和成年大鼠肺胶原蛋白的改变、转化生长因子β(transforming growth factor β,TGF-β)水平的变化、基质金属蛋白酶(matrix metalloproteinases,MMPS)MMP2,MMP9以及基质金属蛋白酶组织抑制因子(tissue inhibitor of metalloproteinases,TIMPS)表达调控的变化,寻找肺间质纤维化发病率随年龄增加的可能原因.方法:通过羟脯氨酸定量测定肺组织胶原含量,ELISA方法测定TGF-β1蛋白水平,荧光定量PCR(realtime PCR)技术测定MMP2,MMP9,TIMP1,TIMP2,TIMP3 mRNA表达量的变化.结果:与成年大鼠组相比,老龄大鼠肺细胞外胶原成分增加,TGF-β1蛋白水平升高[(756±160)pg/g vs(1 000±246)pg/g,P＜0.05],MMP2 mRNA(3.15±1.76 vs 0.17±0.13,P＜0.01)表达水平降低,MMP9 mRNA(1.66±0.67 vs 1.74±0.87,P＞0.05)表达水平没有明显变化.老龄大鼠组TIMP1 mRNA(2.00±1.74 vs 0.11±0.06,P＜0.01),TIMP2 mRNA(7.60±2.51 vs 2.69±1.76,P＜0.01),TIMP3 mRNA(1.32±0.46 vs 0.29±0.16,P＜0.01)表达水平均低于成年大鼠组.结论:肺老化后细胞外胶原比例增加,TGF-β1蛋白水平升高,MMP2和MMP9转录水平改变,TIMPS mRNA转录水平降低等多重因素共同构成肺间质纤维化发病率随年龄增加的基础.     

高能电子线皮肤辐射损伤大鼠自由基的变化

为探讨电子射线放射损伤大鼠局部与全身自由基的改变情况与机理，采用直线加速器局部皮肤照射制作SD大鼠的放射损伤模型，观察全血与局部皮肤MDA、NOS、SOD、GSH-PX含量的改变。结果表明，电子射线损伤后大鼠血清自由基含量发生明显变化，其中MDA、NOS在实验剂量范围内呈上升趋势；SOD、GSH-PX则呈下降趋势。而局部皮肤受照后以上指标虽有改变但呈不规则变化。结果提示，电子射线损伤后能产生自由基，但其作用效应主要在全身，对受照局部皮肤组织自由基改变的作用不明显。     

中波紫外线诱导人皮肤成纤维细胞衰老及机制研究

目的 探讨中波紫外线(UVB)对人皮肤成纤维细胞的诱导衰老作用及可能机制.方法 使用组织块法分离培养原代人皮肤成纤维细胞,通过观察不同剂量UVB照射后的细胞形态学改变以及β-半乳糖苷酶(p-gal)细胞衰老染色情况,确定后续实验诱导皮肤成纤维细胞衰老的UVB单次照射剂量.于经确定剂量UVB照射后的不同时点(12、24、48、72 h)收集细胞及其细胞培养上清液,Western blotting检测诱导衰老的皮肤成纤维细胞衰老相关蛋白P16表达;ELISA法检测诱导衰老细胞Ⅰ、Ⅲ型胶原及基质金属蛋白酶1、3(MMP1、MMP3)表达.结果 经40 mJ/cm2 UVB单次照射的人皮肤成纤维细胞呈现典型的衰老细胞形态学改变,β-gal细胞衰老染色阳性细胞百分比为(88.75±5.32)%,确定诱导衰老UVB单次照射剂量为40 mJ/cm2.Western blotting显示,诱导衰老皮肤成纤维细胞P16蛋白表达随UVB照射时间的延长而明显升高;ELISA法检测发现诱导衰老细胞Ⅰ、Ⅲ型胶原表达随UVB照射时间的延长而明显减少,MMP1和MMP3表达则随UVB照射时间的延长而显著增加.结论 UVB照射能诱导人皮肤成纤维细胞发生衰老,其机制可能与细胞胶原合成减少而降解增加有关.     

新疆紫草脂质体凝胶剂对放射性皮肤损伤的作用

目的考察新疆紫草脂质体凝胶剂对放射性皮肤损伤创面及愈合相关因素的影响。方法用不同体积的脂质加入到适量的凝胶基质中制备脂质体凝胶剂,考察其体外释药特性;以直线加速器G射线照射Wistar大鼠后臀部,建立皮肤放射性损伤模型,观察模型用药后的愈合时间、创面组织病理改变、皮肤羟脯氨酸（hyp）含量、血清MDA、SOD、IL-6、TGF-β变化情况。结果脂质体凝胶剂组（AELG）体外释药曲线符合Higuchi方程,透皮速率比普通凝胶低。40Gy剂量单次照射后20d左右出现脱毛、溃烂现象。经照射后皮肤组织中羟脯氨酸含量,血清中MDA、IL-6、TGF—G水平明显下降,SOD含量显著升高（P〈0．05）。创面用药后,各给药组溃疡4～5w基本愈合,愈合时间较阴性组明显缩短（P〈0．05）。皮肤组织中羟脯氨酸含量所升高,血清中MDA、IL～6、TGF-β含量升高,SOD含量下降,脂质体凝胶剂效果优于普通凝胶剂。结论新疆紫草脂质体凝胶剂对放射性皮肤损伤的愈合有一定的促进作用,其机理可能与促进胶原蛋白合成有关。     

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.     

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.     

安心颗粒对急诊经皮冠状动脉介入术后生活质量影响的研究

目的：评价使用安心颗粒对急诊经皮冠状动脉介入术（PPCI）术后生活质量的影响.方法：将160例接受PPCI的急性ST段抬高型心肌梗死患者随机分为安心颗粒组（术前顿服安心颗粒8.8g,术后安心颗粒4.4 g/次,每日2次）和对照组（仅接受基础药物治疗）.所有患者均服用阿司匹林、氯吡格雷和阿托伐他汀.分别在入院时、出院前1d、出院后180 d时,应用心肌梗死多维度量表（MIDAS）、中文版SF-36评价量表对患者生活质量评分.并观察术后30 d以内的出血并发症、血小板减少症发生情况.结果：入院时和出院前1d,两组患者的心肌梗死MIDAS、SF-36量表评分比较无差异（P＞0.05）;出院后180 d时,与对照组比较,安心颗粒组MIDAS、SF-36评分明显减低（P＜0.05）;组内与入院时比较,两组出院前1d、出院后180 d时,MIDAS、SF-36评分均降低（P＜0.05）.两组患者在随访期间均无大量出血、少量出血、重度和极重度血小板减少症发生,安心颗粒组有4例、对照组有7例发生不明显出血（P＞0.05）.两组发生轻度血小板减少症的患者数比较无差异（P＞0.05）.结论：PPCI使用安心颗粒,能改善急性ST段抬高型心肌梗死患者的生活质量,且不增加出血风险.     

The comparative studies of the influences of Urapidil and Nicardipine on sino-atrial node function, atrio-ventricular node function and hemodynamics

Objective:To investigate the influences of urapidil and nicardipine on rabbit sinus function,atrio-ventricular node function and hemodynamics.Methods:Thirty-two Angora's rabbits were selected and randomly divided into four groups.U1 group:urapidil 0.25 mg/kg;U2 group:urapidil 0.5 mg/kg;N1 group:nicardipine 10 μg/kg;N2 group:nicardipine 20 μg/kg.All these medicine were administrated within 30 seconds.Measurements were taken before and after the administration of urapidil or nicardipine for the following data:mean blood pressure(MAP),heart rate(HR),sino-atrial conduction time(SACT),maximal sinoatrial recovery time(SNRTmax)corrected sinus node recovery time(CSNRT),index of sinus node recovery time(SNRTI),Wenckebach A-V conduction frequency (WB),and P-R interval.Results:Significant MAP and HR changes were identified in all of the four groups before and after administration of both urapidil and nicardipine.No significant changes could be found in the rest of the parameters.Intergroup analysis showed that SACT and CSNRT of N1 and N2 groups were shorter than those of the U2 group(P＜0.01);the MAP decreased(P＜0.01)and the HR increased drastically(P＜0.01).Conclusions:Neither urapidil(0.25 mg/kg,0.5 mg/kg)nor nicardipine(10μg/kg,20μg/kg)has any significant influence on rabbit sinus function or rabbit atrio-ventricular node function.Nicardipine could be a better choice than urapidil for parafunctional sinus node patients.     

Expression of OPGmRNA and ODFmRNA in the patients of hip fracture due to osteoporosis

Objective：To investigate the gene expression of osteoprotegerin（OPG） and osteoclast differentiation factor（ODF） in the bone tissue of patients with hip fracture due to osteoporosis. Methods：OPGmRNA and ODFmRNA in the bone tissue in 50 cases of osteoporosis sufferers（over 50 years old） with hip fracture（Observer Group） and 30 cases of hip facture sufferers with no osteoporosis（Control group） were analyzed with the Semi-Quantitative RT-PCR method. Results：The mRNA expressed of ODF, OPG were both high in the patients with hip fracture. In the control group, the expression of OPG mRNA was observed, while the expression of ODF mRNA was very slight. Conclusion：Aged patients contained all signals including OPG, ODF that are essential for inducing osteoclastogenesis and promoting bone resorption.     

Dynamic Changes in Serum Estradiol and Progesterone levels in Patients of Premenstrual Syndrome with Adverse Flow of Liver-qi

Objective:To probe into the influence of changes of ovarian hormones on the pathogenesis of the specific sub-type premenstrual syndrome(PMS)and reveal partial microcosmic mechanisms of adverse flow of liver-qi.Methods:Estradiol(E2)and progesterone(P)levels in serum were determined at different phases of menstrual cycle by radioimmunoassay.Results:In the group of PMS with adverse flow of liver-qi.the secretive peak value Of E2 and P at the follicular phase significantly decreased,and the secretive peak value at the luteal phase did not come into being.Conclusions:Low E2 and P secretive peak at the follicular phase and absence of secretive peak at the luteal phase is one of the microcosmic mechanisms of PMS with adverse flow of liver-qi.One of the pathophysiologic mechanisms of specific sub-type PMS is probably the continuous low level of E2and P.     

Real-time Three-dimensional Echocardiography in Assessment of Left Ventricular and Right Ventricular Volumes

Real-time three-dimensional echocardiography (RT3DE)is a new ultrasound technique that enables dynamic threedimensional visualization and quantification of the heart in real time. Investigation of feasibility and methodology of RT3DE in determining left ventricular (LV) and right ventricular (RV) volumes, RT3DE was performed in 35 normal adults using Philips SONOS 7500 system with a 2-4 MHz matrix array transducer. The 60°×60° "pyramid" volume database was obtained and analyzed on a TomTec echo workstation. Both LV and RV volumes were calculated with four 3DE methods (i.e. apical 2, 4, 8, and 16-plane) through manually tracing ventricular endocardial borders in end diastole and end systole. Stroke volumes were then calculated. LV volume was also measured by 2DE Simpson's rule using GE VIVID 7 ultrasound machine.     

SCREENING FOR A 21-CHROMOSOME ABNORMALITY IN PREIMPLANTED EMBRYOS OF ELDERLY WOMEN

Increasing maternal age is the only etiological factor unequivocally linked to Down's syndrome in humans. The occurrence rate of newborns with Down's syndrome is about 1/220 in women over 35 years old. However, the occurrence rate in embryos fertilized in vitro, of the elder woman is unclear. Using FISH we screened the number of chromosome 21 in preimplanted embryos of 5 elderly women (average age, 38.4 years) to study the feasibility and necessity of screening trisomy 21 in embryos in patients over 35 years old at the in vitro fertilization (IVF) center.     

Scientific principles and rigorous processes should be followed in developing clinical guidelines for therapeutic interventions of integrative medicine

A clinical guideline for the therapeutic interventions of integrative medicine may be defined as a written document which states a series of recommendations on therapeutic interventions of integrative medicine for a special disease or condition. The guideline may provide assistance to medical professionals in making clinical decisions aimed at improving the clinical outcome of patients and reducing the costs of medical care（~＇4~. Recommendations issued by a guideline should be based on the best available evidence in both Western and Chinese medicine. For fulfilling this purpose, the development of clinical guidelines for therapeutic interventions in the field of integrative medicine should follow scientific principles and undergo a rigorous processes.