耐甲氧西林葡萄球菌耐药性分析

为了解耐甲氧西林葡萄球菌的耐药状况，并为临床预防与治疗提供依据，我们对61株葡萄球菌进行耐甲氧西林及14种常用抗菌素的耐药率测定。结果：耐甲氧西林金黄色葡萄球菌检出率为52.6%；耐甲氧西林凝固酶阴性葡萄球菌检出率为47.6%；耐甲氧西林葡萄球菌对14种抗菌素的耐药率及多重耐药性均高于甲氧西林敏感葡萄球菌。耐甲氧西林葡萄球菌对万古霉素敏感，提示该药可作为临床治疗这类细菌感染的首选药

人胃粘膜活检标本测氨法诊断幽门螺杆菌感染

目的；探讨人胃粘膜Hp感染的检测法。方法：以纳氏测氨法为基础建立了纳氏试剂快速显色检测人胃粘膜中幽门螺杆菌（Hp）感染的方法，临床检测485例因上消化道症状而行胃镜检查的患者。并以细菌培养、粘膜涂片镜检及血清抗Hp－IgGELISA检测结果为对照。结果：该法的敏感性、特异性、阳性及阴性预测值和诊断效率分别为970％，983％，99．4％，91．3％及95．3％。结论；该法检测Hp感染的原理及特点初步分析表明其与传统尿素酶试验有较大的差别，有一定的实用价值

Development actuality and trend of spectroscopy technique

阐述了光谱学光谱的分类,对原子发射光谱、原子吸收光谱、红外吸收光谱、拉曼光谱、X射线吸收光谱、X射线荧光光谱、紫外-可见吸收光谱、分子发光光谱这; 8种光谱方法的原理与发展现状作出了介绍,对光谱分析方法可能用到的数据处理方法进行了简要的概括。最后对光谱学与光谱分析技术的发展趋势进行了归纳。The classification of spectroscopy spectrum was analyzed. The principle; and development actuality for atomic emission spectroscopy,atomic; absorption spectroscopy, infrared spectroscopy,Raman spectroscopy, X-ray; absorption spectroscopy,X-ray fluorescence spectroscopy,UV-Vis; absorption spectroscopy and molecular luminescence spectroscopy were; introduced. The data processing methods that spectral analysis method; may be used were summarized briefly. Finally, the development trend of; spectroscopy and spectral analysis technology was summarized.国家自然科学基金项目; 福建省软科学计划项

Establishment of laboratory animal model of cagA(+) coccoid Helicobacter pylori

目的　建立Hp圆球体动物模型。 方法　通过去营养法来培养cagA(+)Hp圆球体 ,定期用MTT和SCC方法来检测细菌的代谢活性 ,用吉姆萨染色和电镜负染色来观察细菌在不同时期的形态和变化。在圆球体培养的第 7周 ,进行幽门螺杆菌及其圆球体经口感染裸鼠的实验。结果　通过细菌学、尿素酶实验、16SRNA、cagA(+)PCR检测和电镜负染色等方法确定从已喂服幽门螺杆菌及其圆球体的裸鼠胃里分离到的细菌克隆 ,经证实为cagA(+)Hp。 结论　首次在裸鼠上证实了Hp圆球体是Hp“粪—口”传播链上不可缺少的一环。Aim To establish animal model infected with coccoid Hp Methods cagA(+) coccoid Hp was cultured and formed through nutrition deprivation,its metabolism was studied regularly with the methods of SCC and MTT the forms of different time was observed by Giemsa staining and TEM Experiment of infecting nude mice was done with cagA(+) coccoid Hp cultured as mentioned above for 7 weeks and with nomal cagA(+)Hp Result Bacteria isolated from nude mice infected by Hp or its coccoid form were comfirmed to contain cagA(+)Hp with assays of PCR,TEM and Urease Conclusion The results indicate that the coccoid form is a necessary link of Hp's "stool-mouth" infection in nude mic

重组人组织激肽释放酶结合蛋白抗氧化活性及抗凝活性分析

探究Kallistatin蛋白在体外3种细胞模型中的抗氧化活性及其体外抗凝、溶栓活性.首先,在体外构建HBZY-1,HaCaT,HTR8-s/Vneo细胞的H2O2,Fe-HQ氧化应激模型,并将Kallistatin蛋白应用于构建的氧化应激模型中.然后,采用多浓度梯度法筛选最佳氧化剂浓度,利用细胞活性实验检测以上3种细胞的活性,并进行体外抗凝、溶栓实验考察.结果表明:在H2O2模型中,当Kallistatin蛋白浓度为0.6μmol·L-1时,3种细胞活力与对照组(LD-Hanks缓冲液)相近,低剂量蛋白组(0.06μmol·L-1)的细胞活力明显低于高剂量蛋白组细胞;在Fe-HQ模型中,不同细胞中Kallistatin蛋白剂量的作用不同,Kallistatin蛋白具有体外抗氧化活性,不同应激条件下不同细胞对蛋白剂量的依赖性不同;体外抗凝、溶栓实验表明此蛋白具有相关活性.国家重点研发计划资助项目(2016YFE0101700);;国家自然科学基金资助项目(2015J01342

Fast detection of thanatophoric dysplasia type I p.R248C mutation hot spots and rapid prenatal diagnosis of three TD type I high-risk fetuses

目的针对致死性侏儒症I型（thanatophoric dysplasia typeⅠ,TD-Ⅰ）FGFR3基因的突变热点＂p.R248C＂,建立快速特异的酶切鉴定法（restriction endonuelease testing,RE）和扩增受阻突变系统（amplification refractory mutation system,ARMS）/RE法,并应用于后续3例疑似TD-I高危胎儿的快速产前诊断,以及时防止患胎出生,同时为今后的胚胎植入前遗传学诊断（preimplantation genetic diagnosis,PGD）打下良好基础。方法首先,针对突变热点p.R248C突变前后的序列特点,选择合适的内切酶＂Afe I＂,建立酶切鉴定法。其次,设计双错配碱基特异引物结合Apa LI酶切,建立ARMS/RE双重特异鉴定法。对阴性和阳性结果均再用普通引物扩、测的结果进行验证。结果用Afe I酶切FGFR3基因exons（6-7）普通引物的PCR产物（535 bp）,正常对照及非p.R248C突变的TD病例均能被切成255 bp和280 bp 2个片段,而胎1~胎3均切出255 bp、280 bp和535 bp 3个片段。用特异引物E7（p.R248C）扩增,正常对照和非p.R248C突变的TD病例均扩增阴性,无法进一步做酶切鉴定;而p.R248C突变均扩增阳性,当再用Apa LI酶切PCR产物（365 bp）时,胎1~胎3均切出22 bp和343 bp 2个片段。通过引物扩、测结果显示：胎1~胎3均是p.R248C杂合突变。结论该法快速特异、准确可靠,可用于p.R248C突变热点的快速检测及含该突变高危胎儿的快速产前诊断。该法还可用于含p.R248C突变TD-I型家系的PGD。胎1~胎3都是TD-I患胎,建议尽早终止妊娠。Objective To build up the specific rapid methods of RE and ARMS/RE for mutation hotspot＂p.R248C＂in the FGFR3 gene of thanatophoric dysplasia type I（TD-I）,then use the method for rapid prenatal diagnosis of 3 follow-up high-risk fetuses of TD-I to stop the birth of suffering fetuses,at the same time,lay a good foundation of preimplantation genetic diagnosis（PGD） for the future.Methods First,according to the characteristics of the sequence of the mutation hot spot p.248 C before and after mutation,the appropriate enzyme＂Afe I＂was selected to establish the identification method of enzyme digestion.After that,specific primers of double mismatch bases were devised,combining the method of digestion of Apa LI（ARMS/RE） to achieve the double identification.In the end,the sequences obtained by common primers were used to verify the negative and positive results through DNA sequencing.Results For the normal control and the patient without p.R248 C mutation of TD-I,PCR products（535 bp） of exons（6-7） of FGFR3 gene,amplified by common primers,could be digested into 2 fragments（255 bp and 280 bp） by Afe I.However,for fetuses 1~3,the PCR products could be digested into 3 fragments（255 bp,280 bp and 535 bp）.The normal control and the patient without p.R248 C mutation of TD-I were negative using the specific primers E7（p.R248C）,which could not be further identified by enzyme digestion.For the fetuses with p.R248 C mutation of TD-I,PCR products（365 bp） of exon 7 of FGFR3 gene,amplified by specific primers,could be digested by Apa LI and all the digested products were 22 bp and 343 bp.Sequencing results of common primers＇ products indicated：fetuses 1~3 were p.R248 C heterozygous mutation.Conclusions The method is fast,accurate and reliable,which can be available for the fast detection of mutation hot spot p.R248 C and the rapid prenatal diagnosis of high-risk fetus with p.R248 C mutation.This method can also be applied to PGD of high-risk fetus of TD-I with the mutation＂p.R248C＂.Fo闽粤合作科研基金（71010025

A Rapid Method for the Determination of Molar Ratio of Fluorochrome to Protein Techniques by Fluorescence Anisotropy Detection

免疫荧光技术是免疫学检测的重要手段之一，该技术在病原微生物的早期诊断、自身免疫研究、抗原或抗体的免疫组化定位等方面都得到了广泛应用［1］.荧光色素对抗体（或抗原）标记比的测定是免疫荧光技术的重要部分，这是因为标记比合适与否直接关系到检测结果的优劣甚至...The Determination of molar ratio of fluorochrome to protein is an important part in fluorescent antibody techniques.The conventional method is time consuming and with troublesome manipulations.A rapid homogeneous method based on the anisotropy change of the fluorochrome after reacting with protein was presented here.In our experiments, fluorescein isothiocyanate(FITC) and bovine serum albumin(BSA) were chosen to form the FITC/BSA conjugate.A series of solutions containing the two components were prepared and allow to react according to the standard procedure.Fluorescence anisotropy of the mixtures were detected, a diagram of r lg c is then obtained, where c is the concentration of protein.Because FITC in the mixture exists in both free form(F) and binding form(B), the fluorescence anisotropy observed is given by r=f F r F+ f B r B, where r F and r B refer to the anisotropy of free and bound FITC, respectively; and f F and f B represent the fraction of the free and bound forms, respectively, f F+ f B=1.A formula as f B= (r-r F)/( r B- r F) is achieved.Here r B correspods to the value of r in the upper platform region of the r lg c curve, where FITC is bound to protein completely, while r F can be obtained by the determination of anisotropy of FITC in the absence of BSA.So, for each of the mixtures, the binding fraction of FITC can then be calculated.Correspondingly, the content of bound form of FITC that was used to calculate the molar ratio of FITC to BSA can be gained.Since the method avoids the tiresome separation procedure, it bears the merit of time saving and can be used to estimate the molar ratio of fluorochrome to protein rapidly.The determination results of samples by this method were compared with that got from a spectrophotometric analysis.The results were in good agreement.国家自然科学基

苯胺基喹唑啉类酪氨酸激酶抑制剂的电喷雾质谱裂解规律

研究吉非替尼、厄洛替尼和艾克替尼3种苯胺基喹唑啉类酪氨酸激酶抑制剂,在电喷雾质谱正离子模式下的裂解规律.通过电喷雾质谱产生各化合物稳定的[M+H]+准分子离子峰,进而对[M+H]+离子进行高能诱导裂解和碰撞诱导裂解,获得相应化合物的质谱图.结果表明:在电喷雾电离（ESI）多级质谱中,3种药物的裂解主要发生在喹唑啉环C4,C6和C7位取代基上,并伴随分子内重排和H+的迁移重排;在二级质谱图中,吉非替尼的高丰度特征离子质荷比（m/z）为128.1,厄洛替尼的m/z值为278.1和336.1,埃克替尼的m/z值为278.1,304.1.国家自然科学基金资助项目(81302652);;福建省自然科学基金资助项目(2015J01342

PTBP1介导长链非编码RNA RP11-879F14.2发挥抑制心肌纤维化的作用

目的研究长链非编码RNA（lncRNA）RP11-879F14.2调控心肌成纤维细胞纤维化表型的作用及机制。方法对心衰患者及健康对照者心肌组织进行Masson染色检测心肌胶原水平。lncRNA表达谱芯片检测心衰患者及健康对照者心肌中lncRNAs的表达变化，实时定量荧光PCR（RT-qPCR）验证RP11-879F14.2在心衰患者心肌中的表达。利用重组RP11-879F14.2腺病毒（rAd-RP11-879F14.2）感染人心房肌成纤维细胞（HAFs），检测纤维化相关基因Col1a1，Col3a1和Acta2表达。RT-qPCR检测HAFs的核/质组分中RP11-879F14.2的水平。基于生物信息学预测和双荧光素酶报告基因实验鉴定RP11-879F14.2与多聚嘧啶区结合蛋白（PTBP1）的结合作用。检测敲低HAFs中PTBP1表达对RP11-879F14.2调控心肌纤维化相关基因表达的影响。结果Masson染色结果显示，心衰病人心肌组织发生明显纤维化。RT-qPCR结果证实RP11-879F14.2在心衰患者心肌组织中表达增加（P<0.01）。过表达RP11-879F14.2可在RNA及蛋白水平显著抑制心肌纤维化相关基因表达。核质分离及RT-qPCR检测结果证实RP11-879F14.2 主要分布于细胞核中。RP11-879F14.2可与PTBP1结合，并促进HAFs中PTBP1表达，而敲低PTBP1可逆转RP11-879F14.2抑制HAFs中纤维化相关基因表达的作用。结论PTBP1可介导RP11-879F14.2发挥抑制心肌纤维化的作用

A rapid method for the determination of molar ratio of fluorochrome to protein techniques by fluorescence anisotropy detection

The Determination of molar ratio of fluorochrome to protein is an important part in fluorescent antibody techniques. The conventional method is time consuming and with troublesome manipulations. A rapid homogeneous method based on the anisotropy change of the fluorochrome after reacting with protein was presented here. In our experiments, fluorescein isothiocyanate (FITC) and bovine serum albumin(BSA) were chosen to form the FITC/BSA conjugate. A series of solutions containing the two components were prepared and allow to react according to the standard procedure. Fluorescence anisotropy of the mixtures were detected, a diagram of r-lgc is then obtained, where c is the concentration of protein. Because FITC in the mixture exists in both free form(F) and binding form(B), the fluorescence anisotropy observed is given by r = f(F)r(F) + f(B)r(B), where r(F) and r(B) refer to the anisotropy of free and bound FITC, respectively; and f(F) and f(B) represent the fraction of the free and bound forms, respectively, f(F) + f(B) = 1. A formula as f(B) = (r-r(F))/(r(B)-r(F)) is achieved. Here r(B) correspods to the value of r in the upper platform region of the r-lgc curve, where FITC is bound to protein completely, while r(F) can be obtained by the determination of anisotropy of FITC in the absence of BSA. So, for each of the mixtures, the binding fraction of FITC can then be calculated. Correspondingly, the content of bound form of FITC that was used to calculate the molar ratio of FITC to BSA can be gained. Since the method avoids the tiresome separation procedure, it bears the merit of time saving and can be used to estimate the molar ratio of fluorochrome to protein rapidly. The determination results of samples by this method were compared with that got from a spectrophotometric analysis. The results were in good agreement