赖型钩体重组质粒pDJH_2亚克隆及其在大肠杆菌中的高效表达

为了探讨ｐＤＪＨ２的１．９ｋｂ０１７株钩体ＤＮＡ片段可否作为钩体重组疫苗广谱保护性抗原的候选，采用重组ＤＮＡ技术，将ｐＤＪＨ２的１．９ｋｂＤＮＡ片段分别与ｐＴ７－７，ｐＲＳＥＴ系列重组，转化入大肠杆菌进行亚克隆，ＩＰＴＧ诱导表达。结果显示：阳性亚克隆ｐＤＪｔ．ｐＤＪｒＢ１能在大肠杆菌中高效表达；对其进行ＳＤＳ－ＰＡＧＥ分析，可见６８ｋｄ和２３ｋｄ处出现新带，与钩体０１７株外膜同分子量蛋白带位置一致，免疫印迹（特异性抗０１７株外膜抗血清）在６８ｋｄ和２３ｋｄ处分别有印迹；用ｐＤＪｔ亚克隆重组质粒主动免疫豚鼠，可抵抗强毒力株攻击，表现出免疫保护作用。本实验结果提示：（１）ｐＤＪＨ２１．９ｋｂ重组ＤＮＡ片段能以不同质粒为载体，在不同大肠杆菌株中高效表达；（２）该重组ＤＮＡ片段表达产物——６８ｋｄ和２３ｋｄ蛋白，可能是赖型钩体０１７株外膜的保护性抗原。

钩端螺旋体赖型017株外膜脂蛋白LipL41基因的克隆及原核表达

目的 构建钩端螺旋体外膜脂蛋白 L ip L41基因的重组原核表达质粒 ,为进一步制备以 L ip L41为目的基因的亚单位疫苗提供实验依据。方法 根据钩端螺旋体 L.kirscneri RM5 2株外膜脂蛋白 L ip L41基因序列设计引物 ,以赖型钩端螺旋体 0 17株基因组 DNA为模板 ,进行 PCR扩增并 DNA测序 ,以质粒 p GEX1λT为载体 ,插入 L ip L41基因片段构建重组原核表达质粒 ,并检测 L ip L41的表达。结果 测序结果示所得片段为 L ip L41的编码序列 ,酶切及 PCR分析证实重组质粒构建成功 ,SDS- PAGE和 Western blotting分析重组质粒可高效表达蛋白质 L ip L41。结论 L ip L41基因的重组原核表达质粒构建成功 。

问号赖型钩端螺旋体重组质粒pDL121及其表达产物的初步研究

目的 :探讨问号赖型钩端螺旋体重组质粒pDL12 1外源基因及其表达产物 2 3kDa蛋白的特点。方法 :用 6种内切酶对pDL12 1行酶谱分析 ,用Digoxin标记的pDL12 1外源基因片段作探针对不同种属的钩体进行杂交 ,用SDS -PAGE制备 2 3kDa蛋白 ,Westernblot鉴定其免疫原性。结果 :pDL12 1外源基因没有 6种内切酶的酶切位点 ,重组探针与致病性钩体 (serovarlaistrain 0 17,serovarhebdomadisstrain 5 6 6 10 ,serovarpomonastrain 5 6 6 0 8)有杂交信号 ,与非致病性钩体 (serovarpatocstrainPatocI,serovarillinistrain 30 5 5 )无杂交信号 ,亦不识别大肠杆菌。2 3kDa兔抗血清可识别pDL12 1体外表达的 2 3kDa蛋白带和赖型钩体 0 17株超声抗原成份 ;其抗体滴度为 1/12 80 0 ;用pDL12 1细菌裂解液主动免疫豚鼠 ,可使豚鼠抵抗强毒力株钩体攻击。结论 :pDL12 1外源基因可能是赖型钩体的一个新基因 ;该重组探针能鉴别致病性钩体和非致病性钩体 ;2 3kDa抗原有良好的免疫原性 ,可能是赖型钩体 0

组建“三人互赖型合作学习小组”突破高中写作教学困境

针对高中写作教学存在的素材积累、水平提升、修改评价等困境,该文在阐述三人互赖型合作学习小组内涵的基础上,以议论文写作教学为例,从组员互赖积累素材,破除写作前期困境;组员互赖攻关升级,破除写作过程困境;组员互赖评价修改,破除写作后期困境三个方面就组建三人互赖型合作学习小组,破除写作教学困境作探讨。

两株不同毒力株钩端螺旋体37℃培养条件表达谱的差异

目的钩端螺旋体黄疸出血群赖型赖株毒力株56601(中赖)与钩端螺旋体黄疸出血群赖型赖株减毒株(法赖)在遗传背景上具有很大的相似性,但毒力却差别很大。本研究利用钩端螺旋体的cDNA芯片在全基因组水平上研究毒力差别的机制。方法利用钩端螺旋体的cDNA芯片,在37℃培养条件下以钩端螺旋体黄疸出血群赖型赖株毒力株56601(中赖)为测试株,以钩端螺旋体黄疸出血群赖型赖株减毒株(法赖)为对照株,测试了芯片表达谱的变化。结果在37℃培养条件下,毒力株和减毒株的表达谱具有明显的差别。毒力株上调表达的基因有101个,下调表达的有71个。这些差异表达的基因在功能上可分为12类。结论这些差异表达的基因可能在钩端螺旋体毒力株致病力方面发挥一定的作用。

诺氟沙星对钩端螺旋体菌敏感试验

目的 为探讨诺氟沙星抗菌药物对钩端螺旋体黄疸出血群赖型菌株抑制和杀菌的效果。 方法 采用不同浓度的药物 ,进行两种不同方式培养的钩体菌 ,在低倍或高倍镜下观察抑菌及杀菌情况。 结果 用两种不同方式培养钩体菌 ,药物浓度在 0 .0 32～ 0 .0 6 4 mg/ ml剂量下有抑制和杀菌的作用。 结论 在体外培养的钩体菌对诺氟沙星有较好抑杀效果 。

Note on a diffusive ratio-dependent predator-prey model

Subject to the homogeneous Neumann boundary condition, a ratio-dependent predator-prey reaction diffusion model is discussed. An improved result for the model is derived, that is, the unique positive constant steady state is the global stability. This is done using the comparison principle and establishing iteration schemes involving positive solutions supremum and infimum. The result indicates that the two species will ultimately distribute homogeneously in space. In fact, the comparison argument and iteration technique to be used in this paper can be applied to some other models. This method deals with the not-existence of a non-constant positive steady state for some reaction diffusion systems, which is rather simple but sufficiently effective.

螺旋体传染病

问号钩体趋化相关基因特征分析——霍丽等（上海上海第二医科大学基础医学院病理学教研室200025）：《上海第二医科大学学报》，2005，25（1）：33．35[目的：预测问号钩体黄疸出血群赖型赖株的趋化信号传导途径。并深入分析趋化相关基因。方法：使用ProtParam tool,NCBI／Blastp进行蛋白参数分析和结构域分析，使用Clustalw软件进行多序列对比。与大肠埃希菌K-12趋化相关基因进行比较。

A case of steroid-dependent myeloid granulocytic sarcoma masquerading as Crohn's disease

Small bowel tumors and Crohn's disease are common causes of small bowel obstruction.Early stage neoplasms can easily be mistaken for Crohn's disease.Therefore,thorough work-ups including imaging studies and endoscopic evaluation with biopsies are critical for accurate diagnosis.Here we report a case of an otherwise healthy female with progressive onset of multiple,recurrent obstructive symptoms secondary to terminal ileal narrowing who was referred for management of steroid-dependent Crohn's disease.After thorough evaluation,the diagnosis was revised to myeloid granulocytic sarcoma involving the terminal ileum.In this case,a delay in diagnosis can be detrimental for prognosis,as myeloid granulocytic sarcoma is highly predictive of underlying acute myeloid leukemia and needs urgent referral for chemotherapy and/or resection.

Finite element modeling of pavement responses based on stress-dependent properties of asphalt layer

In order to investigate the stress-dependent properties of hot-mix asphalt （HMA）,a dynamic modulus test was conducted on a group of AC-20 specimens at various stress states and loading frequencies,respectively.A user-defined material （UMAT ）subroutine incorporating stress-dependent constitutive model was developed and finite element （FE）simulation was utilized to confirm the validity of the UMAT.A three-dimensional （3D ）FE model for typical pavement structure was established,considering the HMA layer as a stress-dependent material and other layers as linear elastic materials.Periodic load was applied to the pavement model and the pavement responses were calculated,including dynamic modulus distributions,surface deflection,shear stress and tensile strain in the HMA layer,etc.Both test results and FE model predictions indicate that the dynamic modulus of asphalt concrete is sensitive to stress state and loading frequency.Using the nonlinear stress-dependent model results in greater predicted pavement responses compared with the linear elastic model.It is also found that the effects of stress-dependency on pavement responses become more significant as loading frequency decreases.

A time-dependent baroclinic model on NEC bifurcation

As it is well-known, the North Equatorial Current (NEC) bifurcates into the Kuroshio flowing northward and the equatorward Mindanao Current, which is well depicted by Munk's theory in 1950 in terms of its climatology. However, Munk's theory is unable to tell the NEC bifurcation variability with time. In the present paper, a time-dependent baroclinic model forced by wind, in which temporal and baroclinic terms are added to Munk's equation, is proposed to examine the seasonal variability of the NEC bifurcation latitude. An analytical solution is obtained, with which the seasonal variability can be well described: NEC bifurcation reaches its northernmost position in December and its southernmost position in June with a range of about 1° in latitude, consistent with previous results with observations. The present solution will degenerate to Munk's one in the case of steady and barotropic state.

RELATION OF HYPERTENSION TO DIABETIC NEPHROPATHY IN PATIENTS WITH NON-INSULIN-DEPENDENT DIABETES MELLITUS—A PAIR-MATCHED CASE-CONTROL STUDY

To assess the role of hypertension and family history of hypertensio n in the development of nephropathy in patients with non insulin dependent dia betes mellitus (NIDDM).Methods. A retrospective analysis was done on 2 groups of NIDDM patients, one g roup without proteinuria (urine protein< 300mg/24h, n=106) and the other group w ith proteinuria (urine protein≥500mg/24h, n=106). The 2 groups were matched by age(≤±3yrs), sex, ethnic and resident place. Some information of these subject s including demographic; history of disease, family history of diseases, lifesty le and behavior style variables was obtained by questionnaire; some variables w ere measured, including systolic blood pressure (SBP), diastolic blood pressure (DBP), fasting blood glucose (FBG), quantity of protein in 24h urine. Then condi tional logistic regression analysis was performed.Results. Some factors, including history of hypertension, longer duration of hy pertension, higher levels of the past highest SBP and DBP, were independently as sociated with the occurrence risk of diabetic nephropathy (DN). Their correspond ing odd ratios (OR) with 95% confidence intervals (CI) were 2.00(1.17～3.43), 1 .25(1.08～1.46), 1.38(1.15～1.66), and 1.33(1.09～1.62) respectively, but family history of hypertension was not significantly associated with the development o f DN. When the above mentioned relations were respectively adjusted by some pos sible confounding factors, they still existed.Conclusions. History of hypertension, longer duration of hypertension, higher l evels of the past highest SBP and DBP are independent risk factors for DN in Chi nese NIDDM patients.

Pathology of diabetic myocardiac microangiopathy

Objective To investigate the cause of high cardiovascular lethality in patients with diabetics mellitus. Methods Sections from autopsied coronary arteries and myocardium of dead patients with non-insulin-dependent diabetics mellitus and 12 dead control subjects were used for histomorphometric studies. Results The coronary atherosclerotic lesion in diabetics patients was not different in severity fi＇om those in controls. Nor was there difference in number ofmyofibers or diameters of myoeardic fibers and capillaries.But the capillary density and the ratio of capillary number to myocardic fiber number in diabetics group were significeantly reduced compared with control group（P〈0.0 l）,and the capillary basement membrane in the former was significantly thicker than in the latter（P〈0.01）.Conclision The decrease in number of capillaries and the thickening of basement membrane enhance myocardiac vulnerability to further ischemia and hypoxia,which may undelie high lethality of myocardiopathy in diabetic patients.

Superposition model for analyzing the dynamic ground subsidence in mining area of thick loose layer

The dynamic ground subsidence due to underground mining is a complicated time-dependent and rate- dependent process. Based. on the theory of rock rheology and probability integral method, this study developed the superposltlOn model for the prediction and analysis of the ground dynamic subsidence in mining area of thick !oose layer. The model consists of two parts （the prediction of overlying bedrock and the prediction of thick loose layer）. The overlying bedrock is regarded as visco-elastic beam, of which the dynamic subsidence is predicted by the Kelvin visco-elastic rheological model. The thick loose layer is regarded as random medium, and the ground dynamic subsidence, is predicted by the probability integral model. At last, the two prediction models are vertically stacked in the same coordinate system, and the bedrock dynamic subsidence is regarded as a variable mining thickness input into the prediction model of ground dynamic subsidence. The prediction results obtained were compared w^th actual movement and deformation data from Zhao I and Zhao II mine, central China. The agreement of the prediction results with the. field measurements.show that the superposition model （SM） is more satisfactory and the formulae obtained are more effective than the classical single probability Integral model（SPIM）, and thus can be effectively used for predicting the ground dynamic subsidence in mining area of thick loose layer.

Theoretical understanding of electrocatalysis beyond thermodynamic analysis

As the green and sustainable development of human society highly relies on renewable energy,it has been recognized that electrocatalysis is a key technology to this end.High efficient ways of carbon-neutralization(eCO_(2)RR),reverse artificial nitrogen cycle(RANC),and oxygen chemistry(OER and ORR)all can be driven by electrocatalysis.Advanced theoretical study is an important means to fundamentally understanding electrocatalytic reactions.Herein,we review a few significant issues in theoretical electrocatalysis.First,electrochemical barriers and potential effects are essential for a more accurate description of reaction mechanism and activity.Meanwhile,consideration of competitive reaction path is also one of the important aspects,as novel insights and anomalous volcano trend can be obtained.Finally,a microenvironment exerted by confined space can tune the capacitance of electrochemical interface and(electro)chemical potential of proton,resulting in a possibility to improve reaction activity,which opens a new avenue for design of catalyst.

Modeling of time dependent subsidence for coal and ore deposits

Coal and ore underground mining generates subsidence and deformation of the land surface. Those defor- mations may cause damage to buildings and infrastructures. The environmental impact of subsidence will not be accepted in the future by the society in many countries. Especially acceptance of the ground deformations decreases every year there, where the mining regions are densely urbanized, the The only solution is to limit the subsidence or its impact on the infrastructure. The first is not rentable for the mining industry, the second depends on the precise subsidence prediction and good preventing management involved in the mining areas. The precision of the subsidence prediction depends strictly on the mathematical model of the deformation phenomenon and on the uncertainty of the input data. The subsidence prediction in the geological conditions of the raw materials used to be made on the basis of numerical modeling or the stochastic models. A modified solution of the stochastic model by Knothe will be presented in the paper. The author focuses on the precise description of the deposit shape and on the time dependent displacements of the rock mass. A two parameters＇ time function has been introduced in the algorithm.

The Chaotic Real-Exchange-Rate Growth Model

Chaos theory is used to prove that erratic and chaotic fluctuations can indeed arise in completely deterministic models. Chaos theory reveals structure in aperiodic, dynamic systems. A number of non-linear business cycle models use chaos theory to explain complex motion of the economy. Chaotic systems exhibit a sensitive dependence on initial conditions： Seemingly insignificant changes in the initial conditions produce large differences in outcomes. The basic aim of this analysis is to provide a relatively simple chaotic real-exchange-rate growth model that is capable of generating stable equilibria, cycles, or chaos.

Scaling in Rate-Changeable Birth and Death Processes with Random Removals

We propose a monomer birth-death model with random removals, in which an aggregate of size k can produce a new monomer at a time-dependent rate I（t）k or lose one monomer at a rate J（t）k, and with a probability P（t） an aggregate of any size is randomly removed. We then anedytically investigate the kinetic evolution of the model by means of the rate equation. The results show that the scaling behavior of the aggregate size distribution is dependent crucially on the net birth rate I（t） - J（t） as well as the birth rate I（t）. The aggregate size distribution can approach a standard or modified scaling form in some cases, but it may take a scale-free form in other cases. Moreover, the species can survive finally only if either I（t） - J（t） ≥ P（t） or [J（t） ＋ P（t） - I（t）]t ≈ 0 at t ≥ 1; otherwise, it will become extinct.