用分子克隆技术构建D12S391基因座等位基因分型标准物及群体遗传学研究

目的 解决长期困扰短串联重复序列（short tandem repeat,STR）分型上存在的准确性和标准化问题。方法 先用PCR扩增出D12S391基因座的9个等位基因片段，将其插入pUC重组质粒中，经DNA测序分析证实插入片段的结构及大小，用国际标准将插入的等位基因片段进行命名，最后经转染、扩大培养、扩增及再鉴定后，制备出标准的D12S391等位基因分型标准物。结果 应用此法制备出大量的D12S391基因座等位基因分型标准物，并将其用于调查该基因座在德国Mainz地区、日本Miyazaki地区及中国成都汉族、北京汉族、新疆维吾尔族和甘肃回族6个群体中的基因型分布频率。D12S391基因座在各群体中均有较高的多态性，其非父排除概念及个人识别能力分别为0.609-0.786和0.940-0.952。结论 该法制备的STR基因座等位基因分型标准物在法医科学实践中应用价值极高，D12S391基因座是一个非常适合于群体遗传学研究和法医科学应用的遗传标记。     

1H-NMR技术结合多元统计分析鉴别降香及其伪品

目的：采用核磁共振氢谱(¹H-NMR)技术结合多元统计分析建立降香及其伪品的鉴别方法。方法：获得降香及其伪品的¹H-NMR图谱,建立全成分信息并转化为数据矩阵,检测条件为以含0.03%四甲基硅烷(TMS)的氘代二甲基亚砜(DMSO-d₆)为溶剂,恒温298 K(1 K=-272.15℃),脉冲间隔1.00 s,谱宽12 019.23 Hz,扫描数16次,采样时间1.08 s。对降香及其伪品数据矩阵进行相似度考察和层次聚类分析(HCA),采用正交偏最小二乘法-判别分析(OPLS-DA)对数据矩阵进行分析,找出降香与其伪品之间的差异性成分;建立OPLS-DA类别变量值模型,设定降香类别变量值为1,降香伪品类别变量值为0,阈值±0.3,用于市售降香的真伪鉴别;以OPLS-DA得分图确定市售降香中伪品的种类,并通过薄层色谱法(TLC)进行验证。结果：相似度分析与HCA结果表明,降香与其伪品之间差异明显;OPLS-DA发现降香与其伪品显著性差异成分为反式-橙花叔醇;建立的类别变量值模型可成功对市售降香进行真伪鉴别;OPLS-DA得...     

病人知情同意权的几个辩证法问题

以知情同意为核心的病人自主权是医疗服务和现代医患关系新的支撑点。在人体实验、放弃治疗等领域,充分尊重病人的知情同意权具有极大的现实意义。然而,医生的解释说明、医疗干预和亲属代理同意则构成了对知情同意的限制。因此,必须辩证理解和知情同意的内涵,以最大限度地保障病人利益。     

重度颅脑损伤病人康复期智力和记忆力及情感变化

目的探讨重度颅脑损伤病人康复期智力、记忆力及情感等方面的改变。②方法根据格拉斯哥昏迷指数（ＧＣＳ）所规定的等级,对４１例重度颅脑损伤病人进行智力、记忆力能力测验,并与我国常模比较。③结果康复优良组病人的操作智商、语言商数均在正常范围。重残组７５．０％的病人语言商数明显降低,３６．８％轻残组病人和１００％重残病人操作智商降低。④结论严重颅脑损伤病人存在着智力、记忆力和情感障碍,重度残疾病人以操作智商降低为主,康复优良病人常出现智力与记忆力缺陷。     

益骨胶囊对成骨细胞增殖影响的时效及量效关系研究

目的 :运用中药血清药理学方法 ,观察益骨胶囊含药血清对新生大鼠颅骨成骨细胞 (OB)体外增殖的影响 ,并探讨用于OB药效观察的益骨胶囊含药血清制备条件。方法 :雌性 12月龄SD大鼠 80只 ,随机分为生理盐水组、益骨胶囊低剂量 (11 6g/kg)、中剂量 (34 8g/kg)、高剂量 (10 4 4g/kg)组 ,每日灌胃一次 ,连续 12天 ,分别在灌胃后的 0 5h、1h、1 5h、2h采血 ,分离血清 ,用含不同浓度益骨胶囊的血清培养SD新生大鼠颅骨OB ,MTT法检测细胞增殖情况。结果 :末次灌胃后 1 5h采血组的吸收度 (A)值明显高于各组其它时间 (P <0 0 5 ) ;不同剂量各浓度的益骨胶囊含药血清对体外培养的OB增殖均有显著的促进作用 ,其中以高剂量组的稀释比为 2 5 %的血清浓度最佳。结论 :益骨胶囊含药血清对OB增殖具有明显的促进作用 ;益骨胶囊用于OB药效观察的大鼠含药血清制备条件以人等效剂量 9倍连续 12天灌胃、末次灌胃后 1 5h采血 ,血清添加浓度以 2 5 %为宜。     

产后出血的高危因素分析

目的　分析产后出血发生的相关危险因素 ,探讨预防和减少产后出血的措施。方法　对我院1 990年 1月～ 2 0 0 2年 1 2月间住院分娩的产妇出现产后出血的产妇 31 9例 ,对照 6 38例。通过多元Logistic回归分析评价各危险因素的相对危险性。结果　产后出血的发病率为 2 0 9%。相对独立的多个危险因素作用大小的顺位为 :胎盘粘连或植入、前置胎盘、多胎妊娠及胎儿体重≥ 4 0 0 0g、重度妊高征、流产次数≥ 2次、胎盘早剥 ;在阴道分娩的产妇中 ,作用相对独立的危险因素的顺位为胎盘粘连或植入、胎儿体重≥ 4 0 0 0g、妊高征、第三产程 >1 0min、流产次数≥ 2次。结论　避免产前非意愿妊娠以减少前置胎盘、胎盘粘连或植入 ;减少巨大儿、多胎的发生 ,有助于预防和减少产后出血     

重度先兆子痫终止妊娠孕周和方法与围生结局再评估

目的　探讨重度先兆子痫 (severe pre- eclam psia)终止妊娠时间和方法与围生结局的关系。　方法　系统性回顾分析 1988年 9月至 2 0 0 2年 2月我院收治的 30 4例重度先兆子痫终止妊娠时间、方法与围生结局。　结果　孕 33周前胎死宫内发生率及围生儿病死率明显高于孕 33周后者 ,差异有显著性 (χ2 =6 9.89,P<0 .0 0 1;χ2 =10 .5 0 ,P<0 .0 5 )。孕 33周前和 33周后阴道分娩与剖宫产的新生儿死亡率比较 ,差异均无显著性 (P>0 .0 5 )。　结论　重度先兆子痫发生时间和终止妊娠时间与产科结局和围生结局密切相关 ,在 <34孕周发生的重度先兆子痫 ,适时终止妊娠 ,慎行剖宫产 ,有利提高母婴生存质量。     

Positive correlation of insulin-like growth factor-II with proliferating cell index in patients with colorectal neoplasia.

BACKGROUND: Insulin-like growth factor-II (IGF-II) stimulates cell proliferation and is considered a potential risk factor for colorectal cancer. Tumor levels of IGF-II seem to positively correlate with colorectal cancer cell proliferation. This investigation examined the association of circulating IGF-II to the proliferating cell index (PCI) of tumor and matched normal mucosa in patients with colorectal neoplasia. METHODS: Circulating IGF-II level (ng/mL) was determined in the peripheral blood plasma by ELISA. The proliferating cells in tumor or matched normal mucosa were immunohistochemically stained using the primary antibody against Ki-67. Computer image analysis was used and PCI was expressed as the percentage of Ki-67-positive cells/total counted cells. RESULTS: Sixty-four patients were evaluated; 45 had colorectal neoplasia (27 males/18 females; mean age, 66.8 +/- 11.8 years) and 19 had hyperplastic polyps (6 males and 13 females; mean age, 58.4 +/- 14.4 years). Among patients with colorectal neoplasia, blood IGF-II levels were positively correlated with PCI in the matched normal mucosa (r = 0.46, P < 0.05) but not in the tumor. In patients with hyperplastic polyps, blood IGF-II levels were not correlated with the PCI in the polyps. Blood IGF-II levels were higher in colorectal cancer patients with Dukes' C/D stage (P < 0.01) or with positive lymph nodes (P < 0.01). CONCLUSION: Circulating IGF-II positively correlated with PCI in normal colonic mucosa of patients with colorectal neoplasia, suggesting that IGF-II may have a role in initiating the carcinogenic pathway by stimulating cell proliferation. Blood IGF-II was increased in advanced colorectal cancer, indicating that it might enhance colorectal cancer progression and be a useful marker of poor prognosis.     

In vitro effects of active components of Polygonum Multiflorum Radix on enzymes involved in the lipid metabolism

Ethnopharmacology relevance

Raw and processed Polygoni Multiflori Radix (PMR and PMRP) are used in the prevention and treatment of non-alcoholic fatty liver disease (NAFLD), hyperlipidemia or related diseases. In our previous research, 2, 3, 5, 4′-tetrahydroxy-stilbene-2-O-β-d-glucoside (TSG) displayed the most important role in the total cholesterol (TC) lowering effect among all the chemical constituents of Polygonum multiflorum. Emodin and physcion displayed more favorable triglyceride (TG) reducing effects than TSG. However, there are few researches focus on the approach and mechanism of how do Polygonum multiflorum exhibit good lipid regulation activity. The targeted sites of active substances of Polygonum multiflorum are still not clearly elucidated. This research pays close attention to how major chemical components of Polygonum multiflorum affect the TC and TG contents in liver cells.

Materials and methods

In this research, a sensitive, accurate and rapid in vitro model, steatosis hepatic L02 cell, was used to explore target sites of active chemical substances of Polygonum multiflorum for 48 h. Steatosis hepatic L02 cell was exposed to emodin, physcion and TSG, respectively. The contents of four key enzymes in the pathway of synthesis and decomposition of TC and TG were investigated after exposure. Meanwhile, the contents of lipid transfer protein were also tested. The diacylgycerol acyltransferase 1 (DGAT1) controlled the biosynthesis of TG from free fatty acids while 3-hydroxy-3-methyl-glutaryl-CoA reductase (HMG-CoA reductase) limited the biosynthesis of TC. Hepatic triglyceride lipase (HTGL) and cholesterol 7α-hydroxylase (CYP7A) played the key role in the lipolysis procedure of TG and TC.

Results

The synthesis of TC and TG in steatosis L02 cells were apparently increased in the model group compared to the control group. Intracellular contents of HMG-CoA reductase and DGAT1 increased 32.33% and 56.52%, while contents of CYP7A and HTGL decreased 21.61% and 47.37%. Emodin, physcion and TSG all showed down-regulation effects on HMG-CoA reductase, while up-regulation effects on CYP7A. The most remarkable effect on HMG-CoA reductase was found on emodin. Emodin could reduce the DGAT1 content from 438.44±4.51 pg/mL in model group to 192.55±9.85 pg/mL (100 μm). The content of HTGL in 300 μm physcion group was 3.15±0.15 U/mL, which was more significantly effective than the control, lovastatin and fenofibrate group.

Conclusions

TSG could raise the content of CYP7A and then promote the lipolysis of cholesterol. Moreover, TSG also showed the best LDL-reducing effect. Emodin could inhibit HMG-CoA reductase and DGAT1, which were key enzymes in the synthesis of TC and TG. Physcion increased the content of HTGL, and then could boost the lipolysis of triglyceride. At the same time, physcion showed the best VLDL-reducing effect. In view of the above conclusions, we contributed the lipid regulation activity to an overall synergy of TSG, emodin and physcion.     

肌肉注射维生素D3对哮喘儿童血清25-羟基维生素D的影响

目的:探讨儿童哮喘在吸入沙美特罗替卡松同时一次性肌肉注射维生素D37.5 mg治疗前后血清25-羟基维生素D[25(OH)D]的变化。方法:选取2012年10月至2014年6月在成都市妇女儿童中心医院哮喘门诊就诊的哮喘急性发作初治患儿69例,采用随机数字表法分为对照组36例和观察组33例。两组均给予沙美特罗替卡松每次一吸(50μg/100μg),2次/天;观察组仅第1天加用维生素D330万IU肌肉注射一次。两组均在治疗前及治疗3个月后取静脉血清采用化学发光法测定25(OH)D水平。结果:观察组治疗后血清25(OH)D水平升高,虽未达到理想水平,但可纠正哮喘患儿存在的25(OH)D不足或缺乏,与治疗前比较差异有统计学意义(P<0.05);对照组治疗前后血清25(OH)D水平比较差异无统计学意义(P>0.05);治疗后观察组血清25(OH)D水平高于对照组(P<0.05)。结论:对哮喘患儿给予一次性维生素D330万IU肌肉注射可以纠正其存在的25(OH)D不足或缺乏,可能是改善哮喘患儿临床预后的积极补充治疗措施。