Screening Mouse Genes Related to Blastocyst Implantation by Using PCR Subtraction

Objective To identify genes that may be related to embryo implantation Materials & Methods The PCR subtraction technique was applied at implantation and inter-plantation sites on day 4. 5 of pregnancy in mice. Two novel Expressed Sequence Tags (ESTs ), EST8 and EST81 were identified; their expression in tissues was analyzed by Northern blotting, and their full-length cDNAs were synthesized by PCR.Results We found that these two novel ESTs (EST8and EST81) were noticeably expressed in implantation site in the mouse on day 4. 5 of pregnancy. EST8 was expressed at high level in livers and implantation sites of the mice, while at low level in ovaries and inter-plantation sites. EST81 was predominantly expressed in implantation site and ovary, and at low level in all other tissues. Their complete cDNAs, 1 665bp and 1 264 bp respectively, were synthesized by using PCR.Conclusion The two full-length cDNAs were responsible for embryo implantation,and their functions need to be further studied.     

MISE AU POINT D'UNE TECHNIQUE D'ADMINISTRATION IN VIVO DES VECTEURS GENETIQUES DANS LEMUSCLE CARDIAQUE

Objective In order to improve the in vivo gene transfer into the heart muscle, we have designed a ECG-synchronized microinjection system that allows sequential gene delivery to the myocardium.Methods A cannula was introduced into the right carotid artery of the Wistar rat under general anesthesia.With the ECG-synchronized injection during diastole, the genetic vector (Ad CMV lacZ ) infusion was performed with various concentrations( l07 ～ l010pfu ) and different frequency ( the ratio of heart beats per injection from 1: 1 to 4: 1 ). The hearts of the rats were removed after 7 days for histological examination. Results Best results were obtained with a total vector amount of l09 pfu and a good ratio 3: 1 between heart frequency and injection frequency. The transfection efficiency was increased by use of vasodilators and by an increase of vascular permeability. No signs of myocardial ischemia or ventricular arrythmia were observed. Conclusion We have established a novel and safe method for in vivo gene transfer into the heart. Transgene expression suggests that this method may be useful technique to study cardiac function of treat cardiac diseases by means of gene theratpy.     

47例原发性乳腺癌多药耐药MDR1基因表达及其临床意义

[目的]探讨原发性乳腺癌多药耐药MDR1基因的表达及其临床意义.[方法]采用荧光定量RT-PCR法检测47例乳腺癌组织及15例正常对照(包括5例乳腺纤维腺瘤、10例癌旁组织)MDR1基因的表达.[结果]乳腺癌MDR1基因表达阳性率为46.8%,与病人年龄、肿瘤大小、淋巴结转移与否、ER、PR状况、绝经与否无关.对照组中无MDR1基因表达.[结论]乳腺癌MDR1基因的表达可作为乳腺癌化疗耐药的评价指标,能否作为判断预后的独立指标尚需进一步研究.     

Molecular basis of severe myoclonic epilepsy in infancy

Severe myoclonic epilepsy (SMEI) or Dravet syndrome is caused by mutations of the SCN1A gene that encodes voltage-gated sodium channel alpha-1 subunit. Recently, we generated and characterized a knock-in (KI) mice with an SCN1A nonsense mutation that appeared in three independent SMEI patients. The SCN1A-KI mice well reproduced the SMEI disease phenotypes. Both homozygous and heterozygous knock-in mice developed epileptic seizures within the first postnatal month. In heterozygous knock-in mice, trains of evoked action potentials in inhibitory neurons exhibited pronounced spike amplitude decrement late in the burst but not in pyramidal neurons. We further showed that in wild-type mice the Nav1.1 protein is expressed dominantly in axons and moderately in somata of parbalbumin (PV) – positive inhibitory interneurons. Our immunohistochemical observations of the Nav1.1 are clearly distinct to the previous studies, and our findings has corrected the view of the Nav1.1 protein distribution. The data indicate that Nav1.1 plays critical roles in the spike output from PV interneurons and further, that the specifically altered function of these inhibitory circuits may contribute to epileptic seizures in the mice. These information should contribute to the understanding of molecular pathomechanism of SMEI and to develop its effective therapies.     

The role of Self-Defined Race/Ethnicity in Population Structure Control

Population-based association studies are powerful tools for the genetic mapping of complex diseases. However, this method is sensitive to potential confounding by population structure. While statistical methods that use genetic markers to detect and control for population structure have been the focus of current literature, the utility of self-defined race/ethnicity in controlling for population structure has been controversial. In this study of 1334 individuals, who self-identified as either African American, European American or Hispanic, we demonstrated that when the true underlying genetic structure and the self-defined racial/ethnic groups were roughly in agreement with each other, the self-defined race/ethnicity information was useful in the control of population structure.     

高度近视的病因学研究进展

高度近视发生的内因、外因，包括遗传倾向，相关基因定位，巩膜胶原白体免疫学说，视网膜生物活性物质失调学说，环境因素等，本研究就这些因素对高度近视发病的影响做一综述。现统一的认识有：①高度近视具有明显的遗传倾向。②迄今为止，已找到4个高度近视相关基因，它们的遗传方式均为常染色体显性遗传。③高度近视的巩膜组织病理改变和胶原代谢障碍引发了人们对高度近视免疫相关基因的深入研究。已发现HLAⅡ类基因与部分高度近视具有相关性。④视网膜中存在的生物活性物质直接或间接参与了形觉剥夺性近视的形成。⑤环境因素并非高度近视发病的决定因素，它仅起一定程度的促进作用。将今后的工作重点放在高度近视基因定位的研究，基因表达的调控及巩膜胶原代谢、网膜生物活性物质之间的关系研究上，无疑会进一步揭示高度近视的病因，为防治高度近视开辟新途径。     

Recombinant glycoprotein based vaccine for Chandipura virus infection

Chandipura virus (CHPV) has emerged as an important pediatric encephalitis-causing pathogen with very high mortality in India. No specific vaccine or treatment is available till date. We attempted to prepare a candidate vaccine employing recombinant CHPV Glycoprotein (rGp). The Glycoprotein gene (G-gene) of CHPV was expressed using Baculovirus expression system. The rGp was purified by HPLC and used for mice immunization, 3 doses, and 4 weeks apart. One microgram rGp was found to be optimum. Sero-conversion was observed as early as 2nd week by detecting anti-CHPV IgG antibodies. Antibody titres were immunogen-concentration dependent. Intracerebral challenge of the immunized mice with 100 LD₅₀ of the homologous strain demonstrated 90% protection. In in vitro neutralization, antibodies from the immunized mice were able to neutralize heterologous viruses. There was 60% T cell proliferation observed against rGp in immunized mice. The study shows that rGp induces both arms of immune response and represents an ideal vaccine candidate for further evaluations.