致病性奈瑟菌属免疫球蛋白Al蛋白酶最新研究进展

一直以来,免疫球蛋白A1蛋白酶因其对人类IgA1的特异性切割导致黏膜免疫受损而倍受关注。随着生物学和免疫学研究的不断发展,近年来致病性奈瑟菌属IgA1蛋白酶的研究也在不断地深入,尤其是IgA1蛋白酶切割性能与IgA1铰链区变异之间的关系、自身输出的模式以及诱导T细胞免疫等新概念,使得研究者对IgA1蛋白酶有了更进一步的认知。     

黏膜佐剂大肠埃希菌不耐热肠毒素B亚单位优化的淋病奈瑟菌孔洞蛋白B核酸疫苗的构建及其诱导小鼠的免疫应答

Objective To investigate the specific humoral immune response and cellular immune response induced by DNA vaccine with Neisseria gonorrhoeae porin B (PorB) fused with B subunit of Escherichia coli heat-labile enterotoxin B (LTB) in mice. Methods Target genes of porB, ltB and ltB-porB were amplified by polymerase chain reaction (PCR) and cloned into eukaryotic vector pcDNA3.1(-). The recombinants were identified by PCR, enzyme digestion and DNA sequencing.The vectors were transfected into Hela cells, and expressed proteins were checked by cytoimmunofluorescence. Female BALB/c mice were intranasally immunized with recombination vectors. The humoral immune response and cellular immune response were detected by enzyme linked immunosorbent assay (ELISA) and methyl thiazolyl tetrazolium (MTT) colorimetric assay. The expressions of recombination vectors in intranasal mucosal tissues of the immunized mice were detected by immunohistochemistry. The means between groups were compared by analysis of variance. Results All the three recombinants were expressed in Hela cells and intranasal mucosal tissues. The PorB specific IgG in serum and sIgA in vaginal secretions in DNA vaccine immunized mice were significantly higher than those in controls (P＜0.01 ; P＜0.05). Moreover, the sIgA level in pcDNA3.1 (-)/ltB-porB group was higher than that in peDNA3, 1(-)/porB group (P＝0. 002). The levels of interferon-gamma (IFN-γ) and interleukin-4 (IL-4) in the supernatants and stimulation index (SI) of spleen lymphocyte culture in pcDNA3, 1(-)/porB group were (170.04±23.89) pg/mL, (114.68±14.27) pg/mL and 1. 68±0.19, respectively; and those in pcDNA3, 1(-)/ltB-porB group were (161.42±27.50) pg/mL, (124.16±19.04) pg/mL and 1.73±0.28, respectively; which were both higher than those in pcDNA3.1(-)/ phosphate buffered saliae (PBS) group (P＜0. 01; P＜0.05) and pcDNA3.1 (-)/ltB group (all P＜0.05), while there was no significant difference between pcDNA3.1 (-)/ltB-porB group and pcDNA3. 1 (-)/porB group (0. 998, 0. 696, 0. 994; all P＞0.05). Conclusions The constructed DNA vaccines are all successfully expressed in Hela cells and murine intranasal mucosal tissues. The mucosal immunization of the vaccines [pcDNA3. 1 (- )/porB and pcDNA3.1 ( -)/ltBporB] could induce humoral immune response and cellular immune response, especially mucosal immune response. It is confirmed that mucosal adjuvant LTB could promote PorB to induce higher level of mucosal immune response in mice.     

循证药学在行气管切开术中的临床应用

目的：探讨循证药学在临床行气管切开术中的应用。方法：收治气管切开手术患者43例,分为试验组22例和对照组21例。对试验组采取循证药学模式,按照患者病情制定用药指导方案;对照组则按普外科手术后常规用药进行治疗。分析两种试验方法疗效。结果：试验组比对照组手术时间显著缩短、手术出血量显著降低、切口愈合显著缩短、颈部瘢痕明显小。试验组明显高于对照组,有统计学意义（P〈0.01）。两组手术皮下气肿、窒息、切口感染、切口溢痰、低氧血症、气管狭窄等并发症分析及比较,试验组明显低于对照组,有统计学意义（P〈0.01）。且试验组手术成功率高达95.6%。结论：应用循证药学对需行气管切开术的神经外科危重症患者特别适用。     

H因子结合蛋白作为B群流脑候选疫苗抗原的研究进展

脑膜炎奈瑟菌表面蛋白H因子结合蛋白(f HBP),是脑膜炎奈瑟菌的外膜蛋白和重要毒力因子,能与补体调节蛋白H因子结合。结合于脑膜炎奈瑟菌表面的H因子,能下调补体系统的激活作用,使脑膜炎奈瑟菌逃避补体系统的攻击。f HBP能诱导机体产生高水平的保护性抗体,抗f HBP抗体与f HBP结合后可阻止f HBP与H因子的结合,从而使细菌易于被补体系统杀灭和清除。因此,f HBP已成为B群流行性脑脊髓膜炎理想的疫苗候选抗原之一。现就f HBP的结构、基因的表达与调节、作为B群脑膜炎奈瑟菌疫苗候选抗原的作用、应用策略、安全性和有效性等最新研究进展进行综述。     

pcDNA3．1（＋）／NspA真核表达载体的构建及鉴定

目的构建pcDNA3.1( )淋病奈瑟菌外膜蛋白—奈瑟菌表面蛋白A(Neisseria surface Protein A,NspA)基因真核表达载体,为研制淋病奈瑟菌核酸疫苗奠定基础。方法根据淋病奈瑟菌NspA基因序列,设计合成一对引物,用PCR方法从淋病奈瑟菌基因组DNA中扩增出NspA基因片段,将扩增的产物连接于测序载体pUCm-T上,并构建重组体pcDNA3.1( )/NspA,进行酶切、PCR及测序鉴定。结果NspA基因体外扩增产物大小约为525 bp。所构建的pcDNA3.1( )/NspA重组体经双酶切及PCR鉴定,与预期片断的大小相符;测序结果与GenBank中收录的NspA全长序列一致,表明pcDNA3.1( )/NspA真核表达载体构建正确。结论成功地构建了淋病奈瑟菌真核重组表达载体pcD-NA3.1( )/NspA,为NspA蛋白的功能研究和淋病奈瑟菌核酸疫苗的研制提供了物质基础。     

衡阳地区支原体临床分离株对抗生素耐药性监测

目的比较四环素类、大环内酯类和喹诺酮类三类抗菌药物对解脲脲原体(Uu)和人型支原体(Mh)的抗菌作用,为临床用药提供参考依据。方法采用直接肉汤药盘法测定了临床标本中195株Uu和118株Mh对三类抗菌药物中的8种抗生素的敏感性。结果195株Uu对四环素、多西环素、米诺环素、罗红霉素、阿齐霉素、交沙霉素、氧氟沙星、司帕沙星的敏感率分别为21.0%、48.2%、39.5%、79.0%、88.7%、74.9%、28.2%和64.6%。118株Mh对四环素、多西环素、米诺环素、罗红霉素、阿齐霉素、交沙霉素、氧氟沙星、司帕沙星的敏感率分别为5.1%、33.9%、26.3%、0.0%、0.0%、89.8%、70.3%和64.4%。960例混合感染的Uu Mh,对交沙霉素最敏感(79.2%)。结论泌尿生殖道支原体的耐药性监测对指导临床治疗具有重要意义。     

SARS病毒S蛋白部分片段B-细胞表位的多参数预测

目的预测SARS病毒S蛋白(spike glycoprotein)部分片段(aa No.400～600 & aa No.1 100～1 195)的B-细胞表位.方法应用多种参数和方法进行综合分析,包括跨膜分析、保守区分析、同源性比较、Hopp ＆ Woods亲水性参数、抗原性参数、可及性参数、β-转角、万氏法等综合预测方法.结果显示B-细胞识别的表位可能在436～456和1 136～1 146残基或其附近,这两个被预测的表位均含有β-转角和不规则卷曲结构.结论本研究为应用合成肽抗原制备抗SARS病毒S蛋白抗体、诊断非典型肺炎(severe acute respiratory syndrome, SARS)提供了依据.     

利用多媒体技术,改革医学微生物学实验教学

运用多媒体技术,打破医学微生物学实验课程传统的教学模式,改革实验教学,是提高实验教学质量的关键.结合医学微生物学实验的教学特点,分析了运用多媒体技术进行教学改革的优势和应注意的问题,为实验教学改革提供一些经验.     

开展药学服务、控制输液反应

目的控制输液不良反应发生率.方法通过分析输液不良反应发生的直接诱因,人为因素,在输液过程中,开展药学服务,在控制临床输液不良反应方面作出了有益的探索.结果与结论开展药学服务在控制输液不良反应方面具有重要意义.     

重症监护病房铜绿假单胞菌的耐药性分析

目的 了解重症监护病房铜绿假单胞菌的耐药情况,为临床合理用药提供实验室依据.方法 对本院2008年7月～2009年10月各重症监护病房各类送检标本常规进行病原菌培养、鉴定和药敏试验,统计分析铜绿假单胞菌的临床分离株的标本分布及耐药情况.结果 从临床送检的各类标本中分离出铜绿假单胞菌101株,其中痰标本65株,占64.3%,该菌对氨苄西林的耐药率最高(98%),对多粘菌素E的耐药率最低(8.9%),其次对美罗培南的耐药率较低(33.7%),对于其它的抗菌药物均呈现不同程度的耐药.结论 重症监护病房铜绿假单胞菌的感染率很高,其耐药性严重,临床应根据药敏结果合理用药.