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研究一:不同年龄和性别中老年2型糖尿病合并高尿酸血症的特点 研究二:富白细胞-血小板凝胶治疗糖尿病足溃疡作用机制与临床研究
作者: 温天杨
答辩日期: 2013-05-20
导师: 许樟荣
专业: 内科学
授予单位: 北京大学
授予地点: 北京大学解放军306医院教学医院
学位: 硕士
关键词: 糖尿病 ; 2型 ; 高尿酸血症 ; 性别 ; 年龄 ; 富白细胞-血小板血浆 ; 凝胶 ; 糖尿病足溃疡 ; 白细胞 ; 炎性因子 ; 生长因子
其他题名: Study Ⅰ: Clinical characteristics of hyperuricemia in the Chinese patients with type 2 diaebetes----impact of different age and gender Study Ⅱ: A mechanism and clinical study of leucocyte and platelet-rich plasma gel in treatment of diabetic foot ulcer
分类号: R587.1
摘要:

研究一:

目的:了解不同年龄不同性别中老年2型糖尿病合并高尿酸血症的特点。

方法20039月至201112月于解放军306医院就诊的3768例年龄≥40岁的2型糖尿病患者临床资料进行分析,根据尿酸水平将患者分为高尿酸组(A,男性尿酸>420μmol/L,女性尿酸>357μmol/L)和尿酸正常组(B),排序后再进行五分位分组;年龄按40~70岁每5岁分组共分为7组,比较各组高尿酸血症患病率、BMI、腰臀比(WHR)、血压和生化指标。运用Pearson相关分析尿酸与血糖、HbA1c、胰岛素抵抗指数(IR)的相关性,采用logistic回归方法分析尿酸的影响因素。

结果:男性尿酸水平明显高于女性,随年龄变化为先下降后上升;女性尿酸随着年龄增长而上升。男性A组较女性A组平均年龄小、高密度脂蛋白胆固醇(HDL-C)低。女性A组空腹、餐后血糖和男性AHbA1c均低于B组,男女A组空腹、餐后胰岛素、HOMA-IRTGBUNScr均明显高于B组,而HDL-Ce-GFR明显低于B组。男性AAlb/Cr明显高于B组,女性两组无差异。logistic回归分析示男性尿酸与BMITGAlb/Cr呈正相关,与HbA1ce-GFR呈负相关;女性尿酸与BMITGHDL-C呈正相关,与e-GFR呈负相关。 

结论糖尿病合并高尿酸血症的患者有更严重的胰岛素抵抗、血脂异常以及e-GFR下降。男女糖尿病患者的尿酸水平及其年龄、体重、血压等对尿酸水平的影响有所不同。 

研究二:

第一部分  健康志愿者富白细胞-血小板凝胶成分、炎性因子及生长因子的变化

目的:通过测定富白细胞-血小板凝胶中释放的炎性因子IL-1IL-4IL-6和生长因子PDGF-BBTGF-ß1VEGF,观察富白细胞-血小板凝胶的结构,探讨其抗菌作用和促进溃疡愈合的作用机制。

方法:12名健康志愿者,取空腹静脉血经2次离心、分离得到富白细胞-血小板血浆(L-PRP),再以10:1 的比例加入激活剂后混匀得富白细胞-血小板凝胶。将凝胶保存在DMEM培养基中,分别于1371421d后收集含有渗出液的培养基,采用酶联免疫吸附法检测炎性因子IL-1IL-4IL-6和生长因子PDGF-BBTGF-ß1VEGF的浓度。随机选择一名志愿者,制得凝胶后在光镜和扫描电镜下观察凝胶的结构。

结果:富白细胞-血小板凝胶的血小板计数和白细胞总数分别为(813.0±334.5×109/L、(10.5±3.1×109/L,与全血血小板计数(190.0±41.9×109/L、白细胞计数(5.4±1.4×109/L比较差异均有统计学意义(p=0.000)。血小板、白细胞浓缩倍数分别为4.3±1.5)倍、(2.0±0.6)倍,血小板和白细胞回收率分别为(62.4±19.9%、(28.9±7.8%。凝胶中各类白细胞所占比例与全血有所不同,淋巴细胞所占比例最大。IL-10-1d达到峰值,之后逐渐下降。IL-4各时间段释放量无差异。IL-6的释放量在0-1d2-3d4-7d保持较高水平,8-14d15-21d释放量明显下降。PDGF-BBTGF-ß10-1d释放量最大,之后迅速下降。VEGF表现为0-7d逐渐上升,7-21d缓慢下降。光镜和扫描电镜下可见,凝胶中大部分白细胞为淋巴细胞。

结论:富白细胞-血小板凝胶的促进创面愈合的机制除与生长因子PDGF-BBTGF-ß1英文摘要:

Study Ⅰ:

ObjectiveTo evaluate the characteristics of hyperuricemia in the diabetic patients aged ≥ 40 yrs.

MethodsType 2 diabetic patients aged ≥ 40yrs in 306th Hospital were divided into 2 groups based on the blood uric acid (BUA) level, group A with hyperuricemia (> 420 umol/l for male, >357 umol/l for female) and group B with normal BUA level. They were divided into 5 groups based on the BUA level from lowest to highest. These patients were also divided into 7 groups according to age (every 5 yrs differences from 40 to 70). The prevalence of hyperuricemia, BMI, WHR (waist-to-hip ratio), BP (blood pressure) and biochemical parameters were compared among these groups. The spearman analysis was used to analyze the relationship between BUA and glucose, HbA1c, insulin-resistance index. The logistic regression analysis was applied to investigate the related risk factors of hyperuricemia.

ResultsBUA was obviously higher in male than female patients. BUA level was decreased with age and then increased in the male when aged over 65 yrs, while kept increasing with ages for females. In group A, compared with females, males was younger, and with lower HDL-C. Compared with group B, the female patients in group A had lower fasting and post-prandial glucose level, and the males had lower HbA1c. All patients in Group A had higher fasting and post-prandial insulin level, and higher insulin-resistance index than those in Group B. The blood TG, BUN, and creatinine levels were higher and HDL-C and e-GFR were lower in group A than those in group B in both genders. There was significant higher urine Alb/Cr ratio in Group A than Group B for males, but not for females. Logistic regression analysis showed that the BUA was positively related with BMI, TG, Alb/Cr, and negatively related with HbA1c and e-GFR for male, but positively related with BMI, TG, HDL-C and negatively related with e-GFR for female.

Conclusions: The patients with diabetes and hyperuricemia had more severe insulin resistance, lipids disorder and lower e-GFR, but the impacting factors to BUA level different in some way between male and female.

Study Ⅱ:

Part Changes of components, inflammatory factors and growth factors in the leucocyte and platelet-rich plasma gel of healthy volunteers

Objective: To investigate the mechanism of leucocyte and platelet-rich plasma (L-PRP) gel in antibacterial effect and promoting ulcer healing by measuring the concentration of inflammatory factors IL-1, IL-4, IL-6 and growth factors PDGF-BB, TGF-ß1, VEGF released from L-PRP gel and observing the ultrastructure of L-PRP gel.

Methods: Blood samples were collected from 12 healthy volunteers, and L-PRP was obtained by two-step centrifugation, and L-PRP gel was preopared by combining the L-PRP with thrombin and calcium gluconate (10:1). The L-PRP gel was preserved in Dulbecco’s modified eagle’s medium (DMEM), and then the medium containing exudates was collected on days 1, 3, 7, 14 and 21. Quantifications of inflammatory factors and growth factors were performed by using enzyme linked immunoabsorbent assay (ELISA). The ultrastructure of L-PRP gel was observed under the scanning electron microscope (SEM).

Results: The platelet and leucocyte concentration in L-PRP gel were (813.0±334.5)×109/L and (10.5±3.1)×109/L , respectively, and those in whole blood sample were (190.0±41.9)×109/L and (5.4±1.4)×109/L, respectively, with significant difference between L-PRP gel and whole blood sampleP=0.000. The concentration of platelet and leucocyte multiple was (4.3±1.5) times and (2.0±0.6) times , and their recovery rate was (62.4±19.9) %, (28.9±7.8) % , respectively. The leukocyte components were different between L-PRP gel and whole blood sample, and the lymphocyte proportions in L-PRP gel were highest. L-PRP gel released the highest amounts of IL-1 at the first day, and slowly decreased afterward. There were no differences of IL-4 levels among different days. The amount of IL-6 before day 7 was statistically higher than that in other days. The peaks of PDGF-BB and TGF-ß1 emerged at the first day, followed by significantly decreased at later time points. The levels of VEGF increased gradually at first and decreased quietly later. Microscopy and SEM showed that most of leucocytes were lymphocyte in the L-PRP gel.

Conclusions: Besides the expression of growth factors such as PDGF-BB, TGF-ß1 and VEGF, the concentration and component changes of lymphocyte and the release of inflammatory factors such as IL-1, IL-4 and IL-6 might be involved in the mechanism of L-PRP gel in promotions wound healing.

Part Comparison of components, inflammatory factors and growth factors of platelet-rich plasma gel between patients with diabetic foot ulcer and controls

Objective: To investigate the component differences of leucocyte and platelet-rich plasma gel from patients with diabetic foot ulcer and healthy volunteer’ and to measure the factors released from L-PRP gel.

Methods: Blood samples were collected from patients with diabetic foot ulcer and healthy volunteers, L-PRP were obtained by two-step centrifugation, and L-PRP gel was prepared by combining the L-PRP with thrombin and calcium gluconate (10:1). Platelet count, leucocyte count and leucocyte components of L-PRP gel were determined. PDGF-BB, TGF-ß1, VEGF, IL-1, IL-4 and IL-6 released from L-PRP gel were measured using the enzyme-linked immunosorbent assay method. The structure of L-PRP gel was observed by light microscope and scanning electron microscope (SEM).

Results: The platelet and leucocyte count, recovery rate and concentrations of patients’ L-PRP gel were higher than that of healthy volunteers. There were no differences in leukocyte components between patients’ L-PRP gel and whole blood sample, but healthy volunteers’ L-PRP gel had higher lymphocytes and lower neutrocyte proportion than those in whole blood sample. The leucocyte count of patients and healthy volunteers’ L-PRP gel decreased gradually with days passed. SEM showed most of platelets in patients and healthy volunteers’ L-PRP gel had been activated on the day of L-PRP gel prepared, and the contents of platelets in patients’ L-PRP gel were released completely at the end of one week, much earlier than that in healthy volunteers. The peaks of PDGF-BB and TGF-ß1 released from patients’ L-PRP gel emerged at the first day, followed by significantly decreased at later time points. PDGF-BB and TGF-ß1 released from patients’ L-PRP gel were lower than those of healthy volunteers over each period of time, but no statistically significant differences in VEGF levels. VEGF released from patients and healthy volunteers’ L-PRP gel increased gradually at first and decreased quietly later. Patients’ L-PRP gel released the highest amounts of IL-1 at the first day, and decreased afterward. There were no differences of IL-4 levels among days. The amounts of IL-6 released before day 14 was statistically higher than that of the last period. Except IL-6 of 1-3 d, IL-1, IL-4 and IL-6 released from patients’ L-PRP gel were higher than those of healthy volunteers whole period.

Conclusions: There are differences in composition and structure of L-PRP gel and the factors released from L-PRP gel from both patients with diabetic foot ulcer and healthy volunteers. Patients’ L-PRP gel contains more platelet and leucocyte, releases more inflammatory factors such as IL-1, IL-4, IL-6, and less growth factors PDGF-BB and TGF-ß1 than those of healthy volunteers’ L-PRP gel.

Part Ⅲ Typical case report: Treatment of complicated healing-foot-ulcer with leucocyte and platelet-rich plasma gel in patients with diabetes

This part introduced two cases of diabetic foot ulcer treated with leucocyte and platelet-rich plasma gel, including one case of refractory foot ulcer and other with recurrent foot uclcer . The result showed that L - PRP gel resulted in satisfactory efficacy in promoting ulcer healing in those two patients.

语种: 中文
出处: http://xuewei.bjmu.edu.cn/simpsearch.action?keyword=研究一:不同年龄和性别中老年2型糖尿病合并高尿酸血症的特点 研究二:富白细胞-血小板凝胶治疗糖尿病足溃疡作用机制与临床研究&dbid=72
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内容类型: 学位论文
URI标识: http://ir.bjmu.edu.cn/handle/400002259/105438
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作者单位: 北京大学解放军306医院教学医院

Recommended Citation:
温天杨. 研究一:不同年龄和性别中老年2型糖尿病合并高尿酸血症的特点 研究二:富白细胞-血小板凝胶治疗糖尿病足溃疡作用机制与临床研究[D]. 北京大学解放军306医院教学医院. 北京大学. 2013.
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