含手性膦配体2-MBPA铂(Ⅱ)配合物的2DNMR

利用一维和二维ＮＭＲ技术,对含有手性膦配体的铂配合物ｃｉｓ－〔Ｐｔ（２－ＭＢＰＡＨ）２Ｃｌ２〕（１）,ｔｒａｎｓ－〔Ｐｔ（２－ＭＢＰＡＨ）２Ｃｌ２〕（２）,ｃｉｓ－〔Ｐｔ（２－ＭＢＰＡ）２〕（３）和ｃｉｓ－〔Ｐｔ（２－ＭＢＰＡ）（２－ＭＢＰＡＨ）Ｃｌ〕（４）进行１Ｈ和１３ＣＮＭＲ谱分析,区分了化合物（３）和（４）,归属了糖苷部分的１Ｈ和１３ＣＮＭＲ谱线,并根据磷和铂及磷与磷的偶合常数确定化合物（３）和（４）是顺式构型     

含手性膦配体的钯（Ⅱ）配合物的二维核磁共振研究

利用一维和二维ＮＭＲ技术，对含有手性膦配体甲基－３脱氧－３（二苯膦基）－４，６－氧－苄叉基－α－Ｄ一吡喃阿卓糖苷（３－ＭＢＰＡ）和甲基－２－脱氧－２－（二苯膦基）－４，６－氧－苄叉基－α－Ｄ－吡喃阿卓糖苷（２－ＭＢＰＡ）的钯配合物ｔｒａｎｓ—［Ｐｄ（３－ＭＢＰＡＨ）２ＣＩ２」（１），ｔｒａｎｓ－［Ｐｄ（２－ＭＢＰＡＨ）２ＣＩ２］（２）和ｃｉｓ－［Ｐｄ（３－ＭＢＰＡ）２］（３），ｃｉｓ－［Ｐｄ（２－ＭＢＰＡ）２］（４）进行~１Ｈ和~（１３）Ｃ ＮＭＲ谱分析，归属了全部的~１Ｈ和~（１３）Ｃ ＮＭＲ谱线，并根据磷的化学位移及Ｒａｍａｎ谱确定化合物（３）和（４）是顺式构型，对实验中的一些现象也做了简单讨论。     

Synthesis and Crystal Structure of ｛( P Ph_3)_2 Cl Ru( μ- Cl) Ru( P Ph_3) B_(10)H_7〔 O C H( C H_3)_2〕_3｝·0 .4( H_2O)

１　ＩＮＴＲＯＤＵＣＴＩＯＮＴｈｅｆｉｒｓｔｐｏｌｙｈｅｄｒａｌｒｕｔｈｅｎａｂｏｒａｎｅｉｓｓｔｒｕｃｔｕｒａｌｌｙｔｙｐｉｆｉｅｄｂｙｔｈｅｃｏｍｐｏｕｎｄ〔（ＰＰｈ３）２ＲｕＢ１０Ｈ８（ＯＥｔ）２〕ｉｎ１９８４〔１〕,ｗｈｉｃｈｉｓｏｂｔａｉｎｅｄｉｎ４０％ｙｉｅｌｄｂｙｔｈｅｒｅａｃｔｉｏｎｏｆｔｈｅｃｌｏｓｏｄｅｃａｂｏｒａｎｅａｎｉｏｎＢ１０Ｈ１０２－ｗｉｔｈ〔ＲｕＣｌ２（ＰＰｈ３）３〕ｉｎｅｔｈａｎｏｌ／ｃｈｌｏｒｏｆｏｒｍｓｏｌｕｔｉｏｎ．Ｒｅｓｅａｒｃｈｅｒｓｈａｖｅｓｙ…     

烯醇式异桥双铜配合物的合成及磁交换作用的研究

合成了２个异桥双铜配合物：［Ｃｕ２（ｐｆｈｍｐ）Ｂｒ］·２Ｈ２Ｏ（１）和［Ｃｕ２（ｐｆｈｍｐ）Ｃ３Ｈ３Ｎ２］·２Ｈ２Ｏ（２），经元素分析、红外及摩尔电导等手段推知两配合物均为烯醇式构型。变温磁化率测定结果表明，由于外源桥的影响，铜离子间的反铁磁相互作用有明显差别。     

三（3－苯基－4－苯甲酰基异噁唑酮－5）－乙醇－水合铕三元配合物的合成、表征及晶体结构

合成了三（３－苯基－４－苯甲酰基异唑酮－５）－乙醇－水合铕的配合物。有关鉴定和元素分析结果表明：该配合物的分子式为［Ｅｕ（ＰＢＩ）３（Ｃ２Ｈ５ＯＨ）（Ｈ２Ｏ）］·Ｈ２Ｏ，其中３－苯基－４－苯甲酰基异唑酮－５简记为ＨＰＢＩ，是一种新型高酸度的β－双酮。用Ｘ射线单晶衍射法测定了［Ｅｕ（ＰＢＩ）３（Ｃ２Ｈ５ＯＨ）（Ｈ２Ｏ）］·Ｈ２Ｏ的晶体结构，结果表明，该晶体属单斜晶系，Ｐ２１／ａ空间群，晶胞参数：ａ＝１．５５８６（９）ｎｍ，ｂ＝２．１２９７（７）ｎｍ，ｃ＝１．６０４（１）ｎｍ，β＝１１７．９７（５）°，Ｖ＝４．７０３（９）ｎｍ３，Ｚ＝４．中心离子铕为八配位，其配位多面体为畸变的双帽三棱柱。配位的８个氧原子分别来自３个ＰＢＩ配体中的６个氧、１个溶剂化乙醇分子中的氧和１个水分子中的氧，Ｅｕ—Ｏ配位键平均键长０．２３９１（４）ｎｍ。晶体结构中还存在１个未配位的水分子。该水分子、配位水分子、乙醇分子与配体ＰＢＩ中的Ｎ原子之间均存在强的氢键作用。同一ＰＢＩ基团内配位氧之间的平均距离为０．２８０８ｎｍ。     

茜素红S与人血清白蛋白相互作用的分光光度研究

在ｐＨ４．３左右的Ｂｒｉｔｏｎ－Ｒｏｂｉｎｓｏｎ（Ｂ－Ｒ）缓冲溶液中,茜素红Ｓ（ＡＲＳ）与人血清白蛋白（ＨＳＡ）结合,生成红色的复合物,吸光度值与ＨＳＡ含量呈线性关系。复合物的吸收峰值λｍａｘ为５３０ｎｍ,比ＡＲＳ试剂本身红移１１０ｎｍ。其表观摩尔吸光系数ε＝１．０×１０４Ｌ·ｍｏｌ－１·ｃｍ－１。ＨＳＡ标准曲线在其质量浓度１０～９００ｍｇ／Ｌ间呈线性关系,最低检出限为３ｍｇ／Ｌ。研究了该配合物用于蛋白质分光光度法测定的基本条件。应用该法测定了人血清样品,回收率在９６．９％～１０２．１％范围内。实验表明该反应选择性、重现性和对照性均好,操作简便,适用测定浓度范围宽     

三卤代锡钨（钼）双核化合物的制备和晶体结构

ＣｐＭ（ＣＯ）３ＳｎＰｈ３和ＣｐＭ（ｃｏ）３ｓＮｐＨ２ｘ（Ｃｐ＝Ｃ５Ｈ４ＣＨ３，Ｃ５Ｈ４ＣＯＣＨ３；Ｍ＝Ｍｏ，Ｗ；Ｘ＝Ｃｌ，Ｂｒ）与ＨＣｌ或ＨＢｒ发生取代反应，得到三氯、三溴以及混合三卤代锡钨（钼）双核化合物，并经元素分析、＾１Ｈ ＮＭＲ、ＩＲ表征。化合物Ｃｌ３ＳｎＷ（ＣＯ）３Ｃ５Ｈ４ＣＯＣＨ３的晶体属单斜晶系，空间群为Ｐ２１／ｃ，ａ＝０．７６８６（３）ｎｍ，ｂ＝１．２６２２（２）ｎｍ，ｃ＝１．６     

硒半胱氨酸和硒胱氨酸的间接气相色谱测定──溴化氰－气相色谱法

将硒半胱氨酸（ＳｅＣｙｓＨ）甲基化，对硒胱氨酸（ＳｅＣｙｓ）需还原后再甲基化。它们生成的甲基硒半胱氨酸（ＣＨ＿３ＳｅＣｙｓＨ）能与溴化氰（ＣＮＢｒ）发生专一性反应，定量生成的硒氰酸甲酯（ＣＨ＿３ＳｅＮ）可用气相色谱法（ＧＣ）测定。此法简称ＣＮＢｒ－ＧＣ法，检测限４×１０￣（－８）克ＳｅＣｙｓ，准确度８９．５％，相对标准差１２．１％，非含硒氨基酸不干扰。此法适于样品中微量硒氨基酸（硒蛋氨酸ＳｅＭｅｔ，ＳｅＣｙｓＨ和Ｓｅｃｙｓ）的测定。     

硒半胱氨酸和硒胱氨酸的间接气相色谱测定──溴化氰－气相色谱法

 将硒半胱氨酸（ＳｅＣｙｓＨ）甲基化，对硒胱氨酸（ＳｅＣｙｓ）需还原后再甲基化。它们生成的甲基硒半胱氨酸（ＣＨ＿３ＳｅＣｙｓＨ）能与溴化氰（ＣＮＢｒ）发生专一性反应，定量生成的硒氰酸甲酯（ＣＨ＿３ＳｅＮ）可用气相色谱法（ＧＣ）测定。此法简称ＣＮＢｒ－ＧＣ法，检测限４×１０￣（－８）克ＳｅＣｙｓ，准确度８９．５％，相对标准差１２．１％，非含硒氨基酸不干扰。此法适于样品中微量硒氨基酸（硒蛋氨酸ＳｅＭｅｔ，ＳｅＣｙｓＨ和Ｓｅｃｙｓ）的测定。     

三烷基膦－铂－氢配合物的合成及NMR研究

合成了配合物ｔｒａｎｓ－ＰｔＨＸＬ２（Ｌ＝ＰＢｕ３、ＰＰｒ３、Ｐｅｔ３；Ｘ＝Ｃｌ－、Ｂｒ－、Ｉ－、ＳＣＮ－、ＣＮ－），并进行了１ＨＮＭＲ研究。测得Ｐｔ－Ｈ键的１Ｈ在高场的化学位移δＨ＝－８～－２４ｐｐｍ；１Ｊ（Ｐｔ－Ｈ）＝８００～１５００Ｈｚ和２Ｊ（Ｐ－Ｈ）＝１３．０～１７．０Ｈｚ。鉴定了配合物的顺反异构和键合异构。研究了阴离子配体（Ｘ）及中性配体（Ｌ）对Ｐｔ－Ｈ键的δＨ、１Ｊ（Ｐｔ－Ｈ）及２Ｊ（Ｐ－Ｈ）的影响，Ｐｔ－Ｈ键长与Ｐｔ－ＨδＨ的关系。Ｐｔ－Ｈ化学位移与Ｐｔ－Ｈ的伸展振动频率呈线性关系。     

乙型肝炎疫苗预防效果探讨

目的分析研究乙型肝炎疫苗预防效果。方法选取2013年4月—2014年4月中山市三乡医院收治的220例儿童作为研究对象,220例对象全部以经过乙型肝炎疫苗接种,随机分为观察组、对照组,对照组儿童没有给予增强乙型肝炎疫苗接种,观察组儿童则在乙肝疫苗接种的基础之上给予加强乙型肝炎疫苗接种,对两组儿童乙型肝炎疫苗的预防效果进行对比。结果观察组儿童的抗-HBs阳性率和抗-HBc阳性率显著高于对照组(P0.05);观察组儿童的HBs Ag阳性率明显低于对照组(P0.05)。结论乙肝疫苗预防接种免疫人群效果良好,增强乙肝疫苗接种可以有效降低大规模疫苗接种当中不可预知因素对其预防效果的影响,进而使乙肝疫苗预防性接种效果明显提高。     

一种乙肝病毒表面抗原和其单克隆抗体相互作用的SERS研究

利用银纳米粒子的表面增强拉曼散射（SERS）效应，研究了乙肝病毒表面抗原 （HBsAg)和其鼠源单克隆抗体（单抗，Ab-HBsAg)的相互作用。SERS光谱结果表明 ，Ab-HBsAg分子主要通过位于非抗原结合部位的去质子化羧基（COO）^-实现与银 纳米粒子的结合。HBsAg与Ab-HBsAg相互作用形成免疫复合物后，HBsAg分子上的色 氨酸（Trp)残基特征振动完全消失，表明Trp残基位于HBsAg抗原分子的活性区，是 HBsAg与Ab-HBsAg相互作用的重要位点。     

Epidermal cis-urocanic acid levels correlate with lower specific cellular immune responses after hepatitis B vaccination of ultraviolet B-exposed humans

Urocanic acid (UCA) is a major UV-absorbing chromophore in the epidermis and has been suggested to act as one of the initiators of UV-induced immunosuppression. cis-UCA, the isomer from UCA that is formed upon UV exposure, has been shown to impair some cellular immune responses. cis-UCA levels were determined in a study in which the influence of ultraviolet B (UVB) exposure on immune responses after hepatitis B vaccination in human volunteers was established. A significant increase in cis-UCA levels was found in the skin of UVB-exposed volunteers compared with controls. cis-UCA levels, calculated as the percentage of the total UCA amount, in UVB-exposed volunteers correlated significantly with the cumulative UVB dose received in 5 consecutive days, i.e. the higher the UVB dose (J/m2), the higher the cis-UCA levels (until a cis-UCA plateau was reached in the so-called photostationary state). Correlations between skin cis-UCA levels and immune responses were determined, and they revealed no statistically significant correlations among lymphocyte proliferation responses after either mitogenic stimulation or stimulation with recall antigens. No correlation was found between cis-UCA levels and hepatitis B-specific antibody titers. However, we found a statistically significant negative correlation between cis-UCA levels and hepatitis B-specific lymphocyte proliferation responses when volunteers were irradiated with UVB before hepatitis B vaccination. In other words, volunteers with high cis-UCA levels caused by UVB exposure showed lower cellular immune responses against hepatitis B antigen after hepatitis B vaccination.     

A single HBsAg DNA vaccination in combination with electroporation elicits long-term antibody responses in sheep

Vaccines continue to be the most cost effective method to reduce the burden of disease in both human and animal health. However, there is a need to improve the duration of immunity following vaccination, since maintenance of protective levels of antibody in serum or the ability to rapidly respond upon re-exposure (memory) is critical if vaccines are to provide long-term protective immunity. The purpose of this experiment was to test the duration of antibody responses and the ability to generate anamnestic responses following a single immunization with a DNA vaccine encoding hepatitis B surface antigen (HBsAg) delivered by a variety of routes. Sheep immunized with the conventional HBsAg subunit vaccine (Engerix-B) as well as sheep immunized with a HBsAg DNA vaccine, combined with electroporation, generated significant antibody responses that were sustained for 25 weeks after primary immunization. At 25 weeks, all experimental groups received a secondary immunization with the HBsAg subunit vaccine. Sheep that received a primary DNA immunization, in combination with electroporation, mounted an anamnestic response similar to the cohort immunized with the HBsAg subunit vaccine. In contrast, animals immunized with DNA vaccines administered without electroporation elicited no detectable memory response. The presence of immune memory was significantly correlated with the induction of a prolonged primary immune response. Thus, a single DNA vaccination, in combination with electroporation, approached the efficacy of the commercial subunit vaccine in the maintenance of long-term protective serum antibody titres and immune memory.     

红细胞标记抗体的化学发光免疫分析

用红细胞代替辣根过氧化物酶作为双抗体夹心免疫分析中第二抗体的标记物, 建立了一种红细胞标记抗体的免疫化学发光测定乙型肝炎病毒表面抗原的新方法. 在免疫反应完成后, 结合了抗原-抗体免疫复合物的致敏红细胞在低渗溶液中溶血, 释放出血红蛋白. 基于血红蛋白对鲁米诺-H2O2体系化学发光具有催化作用的原理, 采用化学发光法测定血红蛋白含量. 测得的血红蛋白发光强度与待测抗原浓度呈线性关系. 采用这种方法可检测出0.5 ng/mL的乙型肝炎病毒表面抗原. 将该方法与酶联免疫吸附分析(ELISA)结合起来对乙型肝炎患者血清乙肝病毒表面抗原(HBsAg)进行检测, 两者符合率均为97%, 表明本法具有良好的灵敏度和特异性, 可用于临床标本测试.     

UVB exposure impairs immune responses after hepatitis B vaccination in two different mouse strains

Ultraviolet light exposure can impair immune responses that are not restricted to the exposed skin but is also found at other sites, i.e. systemic immunosuppression. Therefore, we investigated the UV-induced modulating effects on vaccination against hepatitis B in a mouse model. Two different mouse strains, BALB/c and C57B1/ 6, were vaccinated intramuscularly against hepatitis B. Mice were exposed to different doses of ultraviolet B (UVB) for five consecutive days on shaved back skin before the vaccination. Vaccination against hepatitis B induced cellular (delayed-type hypersensitivity [DTH] and lymphocyte stimulation test) as well as humoral immune responses in both mouse strains. The DTH responses in C57BB1/6 mice were statistically significantly higher compared with BALB/c mice. UVB exposure induced a dose-dependent suppression of cellular immunity in both strains of mice. C57B1/6 mice seemed to be more susceptible to this suppression. Anti-hepatitis B surface antibodies (total-Ig) were only marginally suppressed after UVB exposure. IgG2a and interferon-gamma levels, both indicators for Th1 immune response, were suppressed in both mouse strains after UVB exposure. In summary, UVB exposure induced a dose-dependent suppression of both cellular and humoral immune responses after hepatitis B vaccination, although the suppressive effects on humoral immunity were limited to IgG2a production. Susceptibility to UVB-induced immunomodulation depended on the strain of mice and their predilection for developing different T cell responses.     

Enhancement of the effectiveness of electroporation-augmented cutaneous DNA vaccination by a particulate adjuvant

DNA vaccines are attracting increased attention due to multiple advantages over conventional vaccines. Attempts to improve these vaccines focus on enhancing DNA delivery and employing novel immunoadjuvants. Electroporation (EP) has emerged as an effective method for delivering DNA vaccines, significantly enhancing humoral and cellular responses. To further improve EP-augmented DNA vaccination, we used micron-size gold particles as a particulate adjuvant. DNA is not bound, or adsorbed, to the particles. Gold particles were coinjected intradermally with plasmid DNA encoding the hepatitis B virus surface antigen (HBsAg) into mice, both in the absence and presence of noninvasive EP. The particles enhanced the percentage of responding animals, and shortened the time for reaching maximal antibody titers by 2 weeks. Subtyping of the produced antibodies revealed a predominantly Th1-like response which did not change significantly with the absence or presence of particles. The particles likely function as an attractant for antigen-presenting cells (APCs), and probably do not affect EP or antigen expression to a significant extent. We conclude that micron-size gold particles injected intradermally together with DNA followed by EP give rise to an accelerated, potent immune response with a strong cellular component. This method may become important for the development of fast-acting therapeutic and prophylactic vaccines.     

Increased in vivo immunological potency of HB-110, a novel therapeutic HBV DNA vaccine, by electroporation

Pulse-induced permeabilization of cellular membranes, generally referred to as electroporation (EP), has been used for years as a tool to increase macromolecule uptake in tissues, including nucleic acids, for gene therapeutic applications, and this technique has been shown to result in improved immunogenicity. In this study, we assessed the utility of EP as a tool to improve the efficacy of HB-110, a novel therapeutic DNA vaccine against chronic hepatitis B, now in phase 1 of clinical study in South Korea. The potency of HB-110 in mice was shown to be improved by EP. The rapid onset of antigen expression and higher magnitude of humoral and cellular responses in electric pulse-treated mice revealed that EP may enable a substantial reduction in the dosage of DNA vaccine required to elicit a response similar in magnitude to that achievable via conventional administration. This study also showed that EP-based vaccination at 4-week-intervals elicited a cellular immune response which was about two-fold higher than the response elicited by conventional vaccination at 2-week intervals. These results may provide a rationale to reduce the clinical dose and increase the interval between the doses in the multidose vaccination schedule. Electric pulsing also elicited a more balanced immune response against four antigens expressed by HB-110: S, preS, Core, and Pol.     

高危儿在顺产与剖腹产中娩出时产生糖代谢紊乱的研究分析

目的分析高危儿在顺产与剖腹产中娩出时血糖的特点,探讨防治糖代谢紊乱路径。方法选取2013年12月至2014年6月于广东省连州市妇幼保健计划生育服务中心产科娩出的85例高危儿为研究对象,按娩出方式分为剖腹产组43例,顺产组42例,应用罗氏血糖检测仪分析高危儿娩出后血糖水平(全血血糖低于2.2 mmol/L诊断为低血糖,全血血糖高于7.0 mmol/L诊断为高血糖),并针对血糖异常制定糖代谢紊乱预防方案,比较两组出生后24 h内的血糖水平。结果顺产组糖代谢紊乱率11.9%显著低于剖腹产组30.2%(P0.05);娩出时剖腹产新生儿脐血的血糖(2.14±0.37)mmol/L明显低于顺产组(2.63±0.59)mmol/L(P0.05),出生后24 h内剖腹产组血糖水平与顺产组比较无显著差异(P0.05)。结论剖宫产高危儿较容易发生血糖异常,出现糖代谢紊乱,故在高危儿出生后应及时监测血糖变化,并采取措施防治血糖异常引起的糖代谢紊乱。     

Selection of recombinant vaccinia viruses (Tian Tan strain) expressing hepatitis B virus surface antigen by using beta-galactosidase as a marker

We constructed a plasmid that contains a small piece of DNA with two vaccinia promoters running in opposite directions--a promoter from a late gene encoding an 11 K polypeptide (P11) and a promoter from an early gene encoding 25K (P25). These promoters were isolated from the Tian Tan strain of vaccinia virus and were flanked by the thymidine kinase (TK) sequence of the same virus. Genes encoding the hepatitis B virus surface antigen (HBsAg) and the Escherichia coli beta-galactosidase (LacZ) were inserted downstream of the 11 K and 25 K promoters respectively so that coexpression plasmids were constructed. Recombinant vaccinia viruses were selected directly by picking blue plaques formed under overlaying agarose medium containing X-gal. HBsAg was expressed to high level by these recombinant viruses. These recombinant viruses showed reduced virulence on rabbit skin and induced anti-HBs after intradermal inoculation of rabbits.