梅毒螺旋体外膜蛋白Gpd的基因型分析及其重组蛋白的免疫原性鉴定

目的　构建梅毒螺旋体 (Treponemapallidum ,Tp)外膜蛋白Gpd基因的原核表达载体 ,检测其表达产物的免疫原性 ,并比较梅毒螺旋体各菌株Gpd基因序列的同源度。方法　从TpNichols株基因组模板中PCR扩增Gpd基因 ,与GenBank登录的序列做blast比较 ,定向克隆构建原核表达重组体pET2 8b( + ) -Gpd ,转入大肠杆菌ril表达菌 ,SDS -PAGE分析重组蛋白的表达 ,Western -blot检测重组蛋白的免疫原性。结果　载体上所连目的基因片段序列与GenBank登录的Nichols株Gpd基因序列完全一致 ,同其他病原性密螺旋体菌株登陆序列比较同源度为 98%～ 1 0 0 %。SDS -PAGE检测诱导产物显示有一Mr约为 41kDa的特异蛋白带 ,免疫印迹技术检测其能与梅毒阳性标准血清反应。结论　Tp原核表达重组体pET2 8b( + ) -Gpd成功构建 ,且能够在ril表达菌中融合表达 ,为进一步研究该蛋白的生物学功能奠定了一定的实验基础。     

Post-operative progress of dystonia patients following globus pallidus internus deep brain stimulation

In the current era of functional surgery for movement disorders, deep brain stimulation (DBS) of the globus pallidus internus (GPi) is emerging as the favoured intervention for patients with dystonia. Here we report our results in 20 patients with medically intractable dystonia treated with GPi stimulation. The series comprised 14 patients with generalized dystonia and six with spasmodic torticollis. Although comparisons were limited by differences in their respective neurological rating scales, chronic DBS clearly benefited both patient groups. Data conveying the rate of change in neurological function following intervention are also presented, demonstrating the gradual but progressive and sustained nature of improvement following stimulation of the GPi in dystonic patients.     

Rapidly progressive tabetic neurosyphilis

Since the sexually transmitted diseases were recognizedas a public health problem in China during the early1980's, the incidence of syphilis has graduallyincreased. Though there have been case reports ofclinical variants of neurosyphilis, including syphiliticcerebrospinal meningitis or meningomyelitis andmeningovascular syphilis, occurring in different regions ofChina, ~(1-3) tabes dorsalis or tabetic neurosyphilis has notyet been reported in China. Here, we report a young manwith rapidly progressive tabetic neurosyphilis admitted toour hospital in October 1999.     

Specific and sensitive diagnosis of syphilis using a real-time PCR for Treponema pallidum

A real-time PCR assay with a Taqman probe was developed that targeted the polA gene of Treponema pallidum. The test was validated using an analytical panel (n = 140) and a clinical panel of genital samples (n = 112) from patients attending a sexually transmitted infections clinic. High sensitivities and specificities of 94-100% were achieved using two real-time PCR platforms, the Rotor-Gene and the iCycler. The assay can be completed within 2 h, enabling reporting in <8 h. This fast and robust assay is suitable for implementation in routine laboratories for diagnosing primary syphilis.     

Environmental effects on the growth and proteolysis of Treponema denticola ATCC 33520

The effect of pH, redox potential, O₂ and H₂ on the growth and proteolytic activity of Treponema denticola ATCC 33520 was studied in a chemostat at different growth rates. The peptidase and protease activities were estimated using different amido-methyl coumarin derivatives and azocasein. The maximum growth rate of T. denticola ATCC 33520 was 0.14 h⁻¹. Reduction of the growth rate of T. denticola by 50–60% gave: an increase in cell mass of 150–200%, a higher acetogenesis and a shift of the pH optimum. The protease and phenylalanine peptidase activities seemed to be of greater importance for the growth of T. denticola ATCC 33520 than the rather low arginine and proline peptidase activities. The redox potential (E_h) played a secondary role. At microaerophilic conditions with 1–5% O₂, the cultures maintained a redox potential below −311 mV and an optimal acetogenesis. The presence of H? induced a marked growth stimulation of T. denticola ATCC 33520. It is concluded that the cell mass and proteolytic activity of T. denticola ATCC 33520 are modulated by the growth rate and the pH and to a lesser extend by the redox potential and presence of O₂. Stagnation of the exudate-flow influences these factors and will lead to an increase of the spirochetal population and proteolysis in the periodontal pocket.     

Injections of -amphetamine into the ventral pallidum increase locomotor activity and responding for conditioned reward: a comparison with injections into the nucleus accumbens

The nucleus accumbens and ventral pallidum receive dopamine (DA) projections from the mesencephalon. Although DA inputs to the nucleus accumbens are implicated in both locomotion and reward processes, little is known of the behavioural significance of DA in the ventral pallidum. These studies examined the effects of -amphetamine injected into the nucleus accumbens or ventral pallidum on locomotor activity and responding for a conditioned reward (CR). In the nucleus accumbens -amphetamine dose dependently (1, 3 and 10 μg) increased locomotion within 5–10 min of injection. Intra-ventral pallidum microinjections of -amphetamine also increased activity in this dose range, but the effect occurred with a longer latency (5–20 min). The magnitude of the response evoked by ventral pallidum injections was lower than that evoked by nucleus accumbens injections. The GABA_A antagonist picrotoxin (0.1 μg) stimulated activity when injected into the ventral pallidum but not the nucleus accumbens, providing a pharmacological dissociation between the two injection sites. In the CR studies, -amphetamine injected into both sites potentiated responding for a CR previously paired with food delivery, without altering responding on an inactive lever. Picrotoxin injected into the ventral pallidum reduced responding and abolished the selectivity of responding for CR. The results show that DA release in the ventral pallidum enhances locomotion and responding for a CR, providing evidence that DA in the ventral pallidum plays a significant role in the mediation of the effects of -amphetamine. The failure of picrotoxin to elevate responding for CR despite increasing locomotor activity indicates that pharmacologically-induced blockade of GABA_A receptors in the ventral pallidum disrupts goal-directed responding.     

PCR检测龈下菌斑中齿垢密螺旋体与牙周组织破坏的关系

目的　建立龈下菌斑标本中齿垢密螺旋体PCR临床快速检测方法 ,了解齿垢密螺旋体感染与慢性牙周炎牙周组织破坏程度之间的关系。方法　 81例慢性牙周炎患者每例采取 2个不同患牙牙位的龈下菌斑标本 ,用终浓度为 1%的TritonX 10 0 10 0℃水浴 10min处理标本以制备DNA模板 ,用PCR检测标本中齿垢密螺旋体特有的相对分子质量 (Mr)为 5 3× 10 3 外膜蛋白基因 (tdpA)扩增片段。结果　 6 7例患者 ( 82 .7% )的 2份龈下菌斑标本齿垢密螺旋体tdpAPCR均为阳性 ,9例患者( 11.1% )的其中 1份龈下菌斑标本可见目的扩增片段 ,5例患者 ( 6 .2 % )的 2份龈下菌斑标本PCR均为阴性 ,16 2例龈下菌斑PCR检测总阳性率为 88.3% ( 143 16 2 )。牙槽骨吸收 >2 3和牙周袋深度≥7mm患牙龈下标本的PCR阳性率较高 (P <0 .0 5 ) ,牙龈指数与PCR阳性率无明显关系 (P >0 .0 5 )。结论　PCR检测tdpA基因可用于慢性牙周炎龈下标本中齿垢密螺旋体的临床快速诊断 ,齿垢密螺旋体感染与牙周组织破坏密切相关。     

Construction of the Eukaryotic Expression Vector for Outer Membrane Protein Tp92 from Treponema pallidum and Its Preliminary Study on the Immune Responses in New Zealand Rabbits

Treponema pallidum subsp. pallidum is the causative a gents of syphilis. Presently, the annual infection rate ofdomestic and international syphilis remains rather high [1,2]. Apart from the serious nature of the disease itself, anumber of studies sugg…     

NMDA receptor antagonist-induced dopamine release in the ventral pallidum does not correlate with motor activation

The ventral pallidum is the output structure of the nucleus accumbens in the ventral corticostriato-thalamocortical loop. Information processing in this loop is critically involved in motor behavior and reinforcement. The ventral pallidum receives a direct dopaminergic input from the ventral tegmental area, but also glutamatergic input from cortical and limbic areas. It has been assumed that dopamine release in the VP is indeed modulated by glutamate. The present study investigated the effects of NMDA receptor blockade on motor behavior and dopamine release in the ventral pallidum. In a first experiment, rats were implanted with microdialysis probes in the ventral pallidum and were systemically injected or locally perfused via the microdialysis probe with dizocilpine (0.32 mg/kg, 10 and 100 microM, respectively). Effects on dopamine and on locomotion were simultaneously monitored. In a second experiment, ventral pallidum was lesioned by quinolinic acid and the effects of systemic dizocilpine (0.08 and 0.16 mg/kg) on locomotion and stereotyped sniffing behavior were determined. It was found that systemic and local dizocilpine administration increased dopamine release in the ventral pallidum to a similar extent whereas only systemic treatment was accompanied by locomotor stimulation. Lesion of the ventral pallidum did not affect locomotion and stereotyped sniffing behavior induced by systemic dizocilpine treatment. Thus, DA release in the ventral pallidum that is elevated by blockade of NMDA receptors is not relevant for activation of motor behavior.