电泳法检测尿液中海洋硫酸多糖聚甘古酯原型物

目的 :建立检测尿液中海洋硫酸多糖类物质聚甘古酯 (911)的电泳方法 ,并用于药代动力学研究。方法 :以氯化十六烷基吡啶 (cetylpyridiniumchloride ,CPC)沉淀法提取尿液中 911,醋酸锌和醋酸钡两步电泳检测尿液中 911,同时测定回收率、电泳测定的检测限。结果 :实验证明该电泳法可以用于检测尿样中的 911原型物 ,911在尿液中的回收率约为 70 % ;水样中 911的检测限为 10 0 μg·L- 1,尿液中检测限为 2 5 0 μg·L- 1,超滤尿中检测限为 5 0 0 μg·L- 1,大鼠口服给药后可以从其尿液中检测到 911原型物。结论 :该电泳法是检测尿液中 911稳定、灵敏的方法     

盐酸达克罗宁凝胶的制备

目的：制备盐酸达克罗宁凝胶。方法：用CMC-Na作基质，制备盐酸达克罗宁凝胶，并建立质控方法。结果：处方设计合理，制备工艺可行，平均回收率为100．4％。RSD=O．8％。结论：该凝胶剂质量稳定可靠，可满足临床使用。     

Utility of peptide–protein affinity complexes in proteomics: identification of interaction partners of a tumor suppressor peptide*

Abstract: We used a N‐biotinylated peptide analog of the C‐terminal domain of the tumor suppressor protein, p21^cip1/waf1 to elucidate peptide/protein interacting partners. The C‐terminal domain of p21^cip1/waf1 protein spanning 141–160 amino acid residues is known to bind PCNA and this interaction is important in many biological processes including cell‐cycle control. This C‐terminal 20‐mer efficiently extracts PCNA in the presence of a variety of N‐ or C‐terminally attached affinity tags. Using difference silver stained 2D gels combined with in‐gel tryptic digests, we identified the difference spots using MALDI‐TOF mass spectrometry‐based peptide mass fingerprinting followed by a database search using profound against NCBIs human nonredundant protein sequence data bank. Identified spots include the p48 subunit of chromatin assembly factor‐1, the heat shock 70 protein analog BiP, calmodulin, nucleolin and a spot similar in size to dimeric PCNA. In contrast, microcapillary ion‐trap LC‐MS/MS analysis of a tryptic digest of entire affinity extracts derived from both control and experimental runs followed by database searches using sequest confirmed the presence of most of the above proteins. This strategy also identified hnRNPA1, HPSP90α, HSP40 and T‐complex protein 1, a protein similar to prothymosin, and a possible allelic variant of the p21^cip1/waf1 protein. The use of N‐biotinylated peptide derived from the C‐terminal domain of p21^cip1/waf1 protein in proteomic analysis exemplified here suggests that peptides obtained from intracellular functional screens could also potentially serve as efficient baits to discover new drug targets.     

微流芯片检测流感病毒多重逆转录聚合酶链反应产物的实验研究

[目的 ]　建立应用微流芯片检测甲 1型、甲 3型、乙型流感病毒多重逆转录聚合酶链反应 (mRT -PCR)产物的方法 ,在mRT -PCR扩增核酸的基础上自动化灵敏的定量检测扩增产物 ,快速检测甲亚型、乙型流感病毒 ,帮助临床快速诊断和鉴别诊断其他呼吸道病毒感染 ,以及明确流感病毒在人群中感染、流行情况。　 [方法 ]　采用经MDCK细胞分离培养的甲 1型、甲 3型、乙型流感病毒毒株病毒液 ,使用 3组特异引物经mRT -PCR扩增核酸 ,扩增产物分别采用毛细管电泳技术经Caliper10 0 0微流芯片分析仪自动化检测和经 2 %琼脂糖凝胶电泳法检测。　[结果 ]　设计三组引物对相应甲 1型、甲 3型、乙型流感病毒靶基因的mRT -PCR扩增产物片段分别为 43 0bp、2 10bp、3 91bp ,本实验mRT -PCR扩增产物经 2 %琼脂糖凝胶电泳 ,与Marker比照 ,DNA条带基本在此位点附近 ;产物采用毛细管电泳法经Caliper10 0 0微流芯片分析仪后 ,分别于 413bp、2 0 3bp、3 79bp处出现陡峭的峰 ,如图所示。　[结论 ]　应用微流芯片采用毛细管电泳法可对流感病毒mRT -PCR产物进行定位及相对定量 ,有助于快速诊断流感病毒感染 ,有助于对流感的监测。     

Molecular epidemiological evidence for ascending urethral infection in acute bacterial prostatitis

PURPOSE: To test the ascending urethral infection in the pathogenesis of acute bacterial prostatitis, we assessed the clonality of Escherichia coli strains isolated from urine and rectal swab of patients with acute bacterial prostatitis using molecular typing methods. MATERIALS AND METHODS: A total of 50 E. coli strains each isolated from urine and rectal swabs of 9 men with acute bacterial prostatitis at diagnosis were examined for 6 urovirulence determinant profiles and pulsed field gel electrophoresis patterns. In 1 case E. coli isolates from the rectal swab of the patient's wife were also examined at diagnosis and after 5 weeks. RESULTS: The urovirulence profile and pulsed field gel electrophoresis demonstrated that causative E. coli was monoclonal in each case, and present in the rectal swab as a predominant (96% to 100%) fecal clone in 2 and a minority clone (2% to 8%) in 4. Furthermore, causative E. coli dominated in the rectal swab of the 1 patient's wife. CONCLUSIONS: Our results are consistent with the ascending route of infection in acute bacterial prostatitis. However, causative E. coli might possibly originate from either intestinal reservoir of the host or household member. Owing to limitations of the cross-sectional design of this study, longitudinal studies are necessary to establish the ascending route of infection in this disease.     

唯地息眼凝胶剂治疗兔芥子气角膜损伤

目的:观察唯地息眼凝胶剂对兔芥子气角膜上皮损伤的治疗作用。方法:新西兰白兔8只,用200μL/L液态芥子气致伤双眼角膜5min,左眼为治疗组,用唯地息眼凝胶剂点眼,4次/d,右眼为对照组,用2.5g/L氯霉素眼液滴眼,4次/d,分别于染毒后2,8,16,24,36,48和72h对角膜荧光素着色区拍照,计算其上皮愈合速率和上皮破损率。结果:治疗组和对照组的角膜上皮愈合速率分别为(1.15±0.19)mm2/h和(1.03±0.12)mm2/h,两者有显著差异(P<0.05),角膜上皮破损率分别为44.4%和54.2%,两者差异显著(P<0.05)。结论:唯地息眼凝胶剂能促进芥子气染毒引起的兔眼角膜损伤的上皮愈合,也能减少角膜上皮反复破损。     

有胫前黑斑的2型糖尿病患者皮肤黑斑和肾活检

目的：研究2型糖尿病(T2 DM)肾脏、皮肤病变的性质及两种组织病变与T2 DM高血糖之间的关系。方法：对573例门诊T2 DM患者进行流行病学调查，了解T2 DM伴胫前皮肤黑斑的发病率；对16例T2DM伴胫前皮肤黑斑的患者均进行了。肾活检和皮肤活检，对101例伴胫前皮肤黑斑T2 DM的血清做了血清蛋白电泳。结果：T2 DM伴胫前皮肤黑斑的发病率为24．78％；肾活检和皮肤活检的病理和免疫组化证实，肾脏有不同的病理形态改变，但肾脏和皮肤存在许多免疫复合物沉积；有4例患者的血清蛋白电泳凝胶上有多余的蛋白带，位置一致，分子质量较大。结论：这类患者肾脏和皮肤同时受自身免疫损害，胰腺β细胞和眼底病变可能为同一发病机制。     

应用微柱凝胶法鉴定婴儿ABO及Rh（D）血型

目的 :比较传统试管法和微柱凝胶法全自动血型系统鉴定婴儿ABO及Rh(D)血型的研究。方法 :选取500份婴儿脐带血标本 ,采用试管法和微柱凝胶法进行ABO和Rh(D)血型 ,并选项取40份抗原性减弱的标本采用两种方法比较。结果 :在鉴定婴儿ABO和Rh(D)血型时两种方法差异无显著性。结论 :微柱凝胶法可取代试管法用于婴儿的ABO和Rh(D)血型鉴定。     

可生物降解的聚乳酸羟基乙酸牙周缓释凝胶的研究

目的采用聚乳酸羟基乙酸(PLGA)研制替硝唑牙周缓释凝胶。方法根据聚合物的分子量、浓度、载药量和增塑剂等因素来优选最佳配方。结果聚合物溶液中N-甲基-2-吡咯烷酮(NMP)P、EG400、PLGA(75∶25)的最佳比例为65∶5∶30,5%替硝唑与聚合物溶液均匀混合即得凝胶。凝胶必须在室温下避光保存。结论该凝胶缓释时间长达7d,用于牙周炎的治疗有效。