Effect of electroacupuncture on the bladder function in rats with bladder hyperactivity

目的：观察电针对排尿频率、骶髓背角血管活性肠肽（VIP）表达的影响,探讨针刺抑制大鼠左旋多巴诱发的膀胱机能亢进的神经生物学机制。方法：将48只SD雌性大鼠按随机数字表分成正常组6只,对照组14只,模型组14只,电针组14只。全部动物均施行膀胱插管术,正常组不做腹腔注射;对照组腹腔注射生理盐水;模型组和电针组动物腹腔内先后注射卡比多巴和左旋多巴;应用连续膀胱测压技术动态观察电针中膂俞对大鼠排尿频率的影响;应用免疫组织化学的方法,观察电针后骶髓背角的vIP动态变化,并用图像分析技术进行定量分析。结果：模型组在腹腔注射左旋多巴后排尿频率较处理前迅速增加（P〈0．05）,电针组在腹腔注射左旋多巴后15～75min时段排尿频率较处理前增加（P〈0．05）,75～105min时段排尿频率接近处理前水平。在腹腔注射左旋多巴后3h时电针组骶髓背角的VIP阳性纤维和终末比模型组、正常组显著增加（P〈0．05）。在腹腔注射左旋多巴后8h时模型组骶髓背角的VIP阳性纤维和终末较正常组显著增加（P〈0．05）,而电针组与正常组则无统计学差异。结论：电针可促进左旋多巴引起的膀胱机能亢进大鼠的躯体传入神经释放VIP,从而降低排尿频率。     

皮肤细胞蛋白质组质谱分析方法的建立及肽质量指纹图谱的构建

目的建立人皮肤细胞蛋白质组学研究所需的质谱分析方法,并获得皮肤细胞蛋白质肽质量指纹图谱。方法体外培养人皮肤成纤维细胞与角质形成细胞,采用固相pH梯度双向凝胶电泳分离其总蛋白质,凝胶经染色后,在图谱上选取2个角质形成细胞蛋白点与1个成纤维细胞蛋白点,进行胶内酶切、基质辅助激光解吸电离飞行时间(MALDI-TOF)质谱分析鉴定。结果获得3个点的肽质量指纹图谱,经Mascot软件搜索人非冗余蛋白质数据库后,鉴定为细胞骨架Actingamma1、tubulinα2及热休克蛋白HSP70。结论皮肤细胞基质辅助激光解吸电离飞行时间(MALDI-TOF)质谱分析技术的建立为皮肤蛋白质组学的进一步研究提供手段。     

β淀粉样肽25-35对原代培养的海马神经元电压门控钙通道电流的影响

目的 探讨β淀粉样肽25-35(Aβ25-35)对海马神经元电压门控的钙通道电流(VGCC)的作用.方法 应用全细胞膜片钳技术记录海马神经元上的VGCC,通过设定不同的钳制电压分别记录高电压激活的钙电流(HVA-ICa)和低电压激活的钙电流(LVA-ICa),并观察Aβ25-35对其的影响.结果 分别对原代培养的海马神经元急性胞外给予2μmol/L和10 μmol/L的凝聚态Aβ25-35,在最大激活电压下加药后ICa幅度分别是加药前的99.80%±0.02%及100.00%±1.58%,差异没有统计学意义.10 μmol/L凝聚态Aβ25-35预孵育24 h可增大ICa,对照组(未给Aβ25-35)为(24.49±4.35)pA/pF,Aβ25-35组(预孵育24 h)为(46.59±7.15)pA/pF,两组差异有统计学意义(P＜0.05);但Aβ25-35组的膜电容[(14.34±1.74)pF]与对照组[(14.44±0.97)pF]相比差异没有统计学意义.结论 急性给予凝聚态Aβ25-35对VGCC没有影响,24 h预孵育凝聚态Aβ25-35增大了VGCC,而膜电容没有改变,提示凝聚态Aβ25-35可通过对细胞膜上钙通道进行分子调控以增大VGCC.     

Utility of B-cell epitopes based peptides of RD1 and RD2 antigens for immunodiagnosis of pulmonary tuberculosis

Tuberculosis (TB) continues to be a major health problem due to lack of accurate, rapid, and cost-effective diagnostic tests. Serodiagnostic tests incorporating highly specific region of difference (RD) antigens (early secretory antigenic target 6 [ESAT-6], culture filtrate protein 10 [CFP-10], culture filtrate protein 21 [CFP-21], and mycobacterial protein from species tuberculosis 64 [MPT-64]) have recently been shown to be promising for specific diagnosis of TB in our lab. However, only few studies have reported the use of synthetic peptides of RD antigens, and none has used them to differentiate TB from sarcoidosis, a close mimic of smear-negative pulmonary TB (PTB) with entirely different management. The present study was conducted with an aim to study the utility of B-cell epitopes based peptides of RD1 (ESAT-6, CFP-10) and RD2 (CFP-21, MPT-64) antigens for immunodiagnosis of PTB for which sputum smear-positive PTB patients, sputum smear-negative PTB patients, sarcoidosis patients, and healthy controls (n = 24/group) were recruited. Bioinformatic software Bcepred was used to predict linear B-cell epitopes, using physico-chemical properties on a non-redundant dataset. Seven peptides as representative B-cell epitopes of ESAT-6, CFP-10, CFP-21, and MPT-64 were evaluated as targets of the antibody responses in TB patients and controls by enzyme-linked immunosorbent assay (ELISA). The current study showed sensitivity with individual peptides ranging from 37.5% to 83.3% for smear positive, 25% to 58.3% for smear negative as compared to 4.16% to 20.8% for sarcoidosis. Four out of 7 peptides that showed higher reactivity with TB patients and better discrimination from sarcoidosis patients representing ESAT-6, CFP-10, CFP-21, and MPT-64 were selected for multiepitope ELISA. The combination of peptides yielded 83.3% sensitivity for smear positive, 62.5% for smear negative, and only 4.16% for sarcoidosis. The specificity, however, for all the peptides/combination was 100%. Combination of peptides has proven to be better than individual peptides as per the latest criteria of the World Health Organization according to which a test that can replace smear microscopy with sensitivity of >90% for smear-positive patients and >65% for smear-negative TB patients with a specificity >95%, and thus, the present study suggests that a test based on combination of peptides selected from mycobacterial RD1 and RD2 antigens could be important for promoting an early diagnosis and management of otherwise difficult to diagnose smear-negative PTB patients. Moreover, it can also be used to discriminate sarcoidosis from PTB, thus preventing the misdiagnosis and mismanagement.     

Using microemulsion formulations for oral drug delivery of therapeutic peptides

Peptide drugs are increasingly becoming a very important class of therapeutic agents with the rapid advances in the field of biotechnology engineering. However, these drugs are generally not suitable for oral administration. In this review, the main physico-chemical and biopharmaceutical characteristics of peptides are summarized. The obstacles to peptide drug absorption and the different possibilities for solving these difficulties are listed. Results using this formulation approach for oral drug delivery of peptides are apparently promising with some specific peptides such as cyclosporin. Various mechanisms are only beginning to be understood and further investigations need to be performed in this area to explain the results obtained with some peptides.     

反刍动物小肽的营养研究进展

肽作为蛋白质的主要消化产物，在氨基酸消化、吸收和代谢中起着重要作用。本文通过对反刍动物小肽的产生与降解、小肽的吸收与营养以及肽的调控技术等方面情况的综述，勾画出了当前反刍动物营养学研究领域内肽营养的研究现状与发展方向。     

阻断CXC趋化因子受体3途径对同种异体胰岛移植急性排斥反应影响的实验研究

目的研究CXC趋化因子受体3（CXC chemokine receptor 3,CXCR3）反义肽核酸（peptide nucleic acid,PNA）对同种异体胰岛移植急性排斥反应的影响。方法实验模型为小鼠同种异体胰岛移植模型。分为生理盐水对照组,PNA CXCR3组和随机错配PNA组。体外T细胞增殖应答能力通过淋巴细胞增殖反应来评估。应用RT-PCR和Western blot检测CXCR3 mRNA和蛋白表达水平。应用流式细胞仪测定脾CD3^＋T细胞CXCR3表达水平。结果与生理盐水组[（6．72±1．48）d]和随机错配PNA组[（6．54±0．86）d]相比,PNA CXCR3组[（9．70±1．57）d]受体有功能胰岛移植物存活时间明显延长。移植后第7天,PNA CXCR3组CXCR3 mRNA水平（1．06±0．07）同生理盐水对照组（1．98±0．22）和随机PNA组（1．87±0．10）比较明显下调（P〈0．01）。PNA CXCR3组移植物CXCR3蛋白水平以及小鼠淋巴细胞增殖能力与另两组比较明显降低（P〈0．01）。结论PNA CXCR3通过降低T淋巴细胞活性来延长同种异体胰岛移植物存活时间,在抑制同种异体急性排斥反应中具有潜在的治疗效应。     

Utility of the Guy’s Stone Score in predicting different aspects of percutaneous nephrolithotomy

Objective

To evaluate Guy’s scoring system (GSS) as a grading system for complexity of kidney stone before percutaneous nephrolithotomy (PCNL) as a predictor for different items of outcome.

TGFα和EGF对前列腺癌细胞系EGFR表达的调控作用

目的：探讨转化生长因子（TGF）α和表皮生长因子（EGF）对前列腺癌细胞系中表皮生长子受体（EGFR）表达的调控作用。方法：采用RT－PCR和Western印迹法分别对TGFα和EGF刺激前列腺癌细胞系LNCaP、PC3、ARCaP后EGFR mRNA表达及其蛋白水平进行定量分析。结果：EGF引起前列腺癌细胞系LNCaP、PC3、ARCaP的EGFR mRNA升高2－5倍,TGFα引起各细胞系EGFR mRNA升高2－10倍。TGFα使各细胞系EGFR不同程度升高;EGF处理的LNCaP EGFR蛋白水平略升高,PC3、ARCaP EGFR蛋白水平降低。结论：TGF／EGF-EGFR通路在前列腺癌发生发展中起重要作用;非依赖型前列腺癌中TGFα－EGFR自分泌环的作用可能强于EGF－EGFR自分泌环。     

普萘洛尔及其对映体血药浓度对血浆心钠素水平的影响

本文研究８名健康志愿者口服单剂量普萘洛尔（４０ｍｇ）和１２名健康志愿者口服多剂量普萘洛尔（８０ｍｇ／ｄａｙ×３）后；静息状态和踏车兴奋状态时血浆心钠素（ＡＮＰ）水平的变化。口服单剂量普萘洛尔后３ｈ，静息状态时血浆ＡＮＰ水平升高１９．４±１１．１ｎｇ／Ｌ（Ｐ＜０．０５）；而运动状态时血浆ＡＮＰ升高更为明显，为６４．８±４０．９ｎｇ／Ｌ（Ｐ＜０．０５）。血浆ＡＮＰ的变化与普萘洛尔对映体Ｓ（－）－ＰＰＬ和Ｒ（＋）－ＰＰＬ浓度均具有良好的线性相关性，相关系数分别为０．８５６３和０．７１９２。口服多剂量普萘洛尔停药后１２ｈ，血浆ＡＮＰ仍可维持较高水平，较基础对照值升高３９．９±２８．３ｎｇ／Ｌ（Ｐ＜０．０５）。此结果提示，普萘洛尔参与了血浆ＡＮＰ水平的调控，这种作用尚难以用β肾上腺素能效应进行解释，可能与抑制ＡＮＰ的体内降解过程有关。