脐血CD34+细胞体外定向诱导分化为T淋巴细胞的实验研究

目的：建立利用人造血干／祖细胞体外定向诱导分化为T淋巴细胞的方法，为研究T细胞生物学特性及细胞免疫提供技术平台。方法：MACS方法分离人脐带CD34^ 细胞接种到人胎儿胸腺基质单层细胞上，IMDM液体培养基含20％人AB血清并加入FL、IL-12、IL-7和IL-2细胞因子组合，于培养7、14、21、28、35、42天取非贴壁细胞利用流式细胞仪对细胞表型进行检测，并进行细胞形态学分析。结果：2周后，CD4^ CD8^ 非成熟T淋巴细胞占细胞总数的0．3％-13．3％，4-5周CD4^ CD8^ T淋巴细胞达到高峰占16．6％-26．5％，且CD3^ CD4^ CD8^ 和CD3^ CD4^-CD8^ T淋巴细胞逐渐增多，6周后达26．5％～64．9％和11．6％-38．9％。培养成熟的T淋巴细胞经PHA IL-2刺激后瑞氏染色鉴定可见大原始淋巴细胞存在。结论：利用人脐血CD34^ 在体外人胎儿胸腺基质单层细胞上加FL、IL-12、IL-7和IL-2细胞因子组合条件下，可诱导分化出T淋巴细胞，并且培养的T细胞对有丝分裂素刺激有增殖反应。     

Histochemical study of RNA content of neurones in the dorsal lateral geniculate nucleus during postnatal development.

Nuclear and cytoplasmic dLGN neurons were investigated by cytophotometric measurements of RNA. This study has been carried out in rats from birth to adulthood. In order to quantify the RNA content a cytophotometer was used. Extinction mean values were obtained which indicated RNA concentrations per surface unit. The nuclear and cytoplasmic surface were calculated simultaneously and from the product of the mean extinction and the surface the RNA total content was calculated. Our results have suggested that the changes are age-related. From day 1 to day 21 the neuronal size and RNA content increase; this may somehow be involved with the differentiation process. Around post-natal day 21 neuronal maturation may begin, reaching its optimal phase around day 42, on which the RNA concentration per surface unit, surface neuronal content and RNA total content are stable.     

人骨髓间质干细胞和嗅鞘细胞移植对大鼠急性脊髓损伤功能修复的比较研究

目的对比人骨髓基质细胞(BMSCs)与人胚嗅鞘细胞(OECs)移植对大鼠脊髓损伤功能修复的影响。方法将45只SD大鼠分成脊髓损伤后BMSCs移植组(BMSCs组)、OECs移植组(OECs组)和PBS对照组(PBS组),通过BBB评分、运动诱发电位(MEP)评估脊髓传导功能的改善状况,免疫组织化学方法检测移植细胞存活和分化情况,病理形态学方法观察组织结构修复情况。结果BMSCs组BBB评分高于OECs组(P<0.05);两细胞治疗组MEP潜伏期明显缩短(P<0.05);BMSCs组嗜银染色可见脊髓损伤近端有较多再生纤维向远端延伸,形成神经纤维束,而OECs组再生纤维较少;两种移植细胞均可在损伤处部分存活,BMSCs组可见BMSCs来源的细胞Nestin、NF、GFAP的阳性表达。结论BMSCs移植比OECs移植能更有效促进大鼠急性脊髓损伤修复。     

慢性束缚应激致大鼠持续性高血糖症与孤束核损伤有关

目的 探讨孤束核联合亚核前部（acNTS）损伤在慢性束缚应激（CRS）所致的胰岛素抵抗性高血糖症发生中的作用。 方法 采用CRS大鼠模型（n=20;7 d束缚+3 d自由活动，共40 d），检测葡萄糖代谢相关指标。 结果 CRS导致约1/3的个体（n=7）持续性的中度胰岛素抵抗性高血糖（空腹血糖不超过11 mmol/L）。CRS高血糖鼠acNTS可观察到神经元染色浓缩，Caspase-3表达和TUNEL阳性染色，提示出现神经元凋亡样改变。机械损伤acNTS（n=6），空腹血糖水平逐渐升高，也能导致胰岛素抵抗性高血糖, 且高胰岛素血症、胰岛平均体积增大和血清皮质酮水平不变等特点与CRS小鼠一致。 结论 CRS损伤了acNTS的葡萄糖敏感神经元，从而使血糖调节紊乱。     

In search of candidate genes critically expressed in the human endometrium during the window of implantation

BACKGROUND: In this prospective randomized blinded clinical trial, we examined gene expression profiles of the human endometrium during the early and mid-luteal phases of the natural cycle. METHODS: An endometrial biopsy was performed on day 16 (LH +3) or on day 21 (LH +8), followed by RNA extraction and microarray analysis using an Affymetrix HG-U95A microchip. Data analysis was carried out using pairwise multiple group comparison with the significance analysis of microarrays (SAM) software. RESULTS: With a false discovery rate of 0, the analysis revealed that 107 genes were significantly and differently expressed (> or =2-fold) during the early versus the mid-luteal phase of the cycle. Forty-five of these genes have not been previously linked to endometrial receptivity. Validation of the microarray data was accomplished using semiquantitative RT-PCR. We demonstrated the presence of estrogen and progesterone response elements (ERE and PRE) by analysis of the 5'-flanking regions of a subset of differentially regulated genes. CONCLUSIONS: Using a strict bioinformatics approach of microarray data, we demonstrated significant changes in candidate genes during the transition of the early to the mid-luteal phase of the human endometrium that may have functional significance for the opening and maintenance of the window of implantation.     

A malformed child with a recombinant chromosome 7, rec(7) dup p, derived from a maternal pericentric inversion inv(7)(p15q36).

We report a child with facial dysmorphic features, hypoplasia of the external genitalia, intestinal malrotation, congenital cardiac defect, and minor limb anomalies. Chromosome studies showed a recombinant chromosome 7, rec(7) dup p, resulting from a maternal pericentric inversion inv(7)(p15 q36). Thus, this child had partial trisomy 7p in addition to a small distal monosomy 7. The clinical findings are compared with those found in previous reports of trisomy 7p. Finally, some general principles for genetic counselling are discussed.     

Identification and characterization of a conserved,stage-specific gene product of Plasmodium falciparum recognized by parasite growth inhibitory antibodies

We have identified a novel conserved protein of Plasmodium falciparum, designated D13, that is stage-specifically expressed in asexual blood stages of the parasite. The predicted open reading frame (ORF) D13 contains 863 amino acids with a calculated molecular mass of 99.7 kDa and displays a repeat region composed of pentapeptide motives. Northern blot analysis with lysates of synchronized blood stage parasites showed that D13 is highly expressed at the mRNA level during schizogony. The first N'-terminal 138 amino acids of D13 were expressed in Escherichia coli and the purified protein was used to generate anti-D13 monoclonal antibodies (MAbs). Using total lysates of blood stage parasites and Western blot analysis, these MAbs stained one single band of approximately 100 kDa, corresponding to the predicted molecular mass of ORF D13. Western blot analysis demonstrated further that D13 is expressed during schizogony, declines rapidly in early ring stages and is undetectable in trophozoites. D13 protein is localized in individual merozoites in a distinct area, as demonstrated by indirect immunofluorescence analysis. After subcellular fractionation, D13 was confined to the pelleted fraction of the parasite lysate and its extraction by alkaline carbonate buffer treatment indicated that D13 is not a membrane-integral protein. Inclusion of certain anti-D13 MAbs into in vitro cultures of blood stage parasites resulted in considerable reduction in parasite growth. The N'-terminal domain encompassing 158 amino acids is 94 and 95%, respectively, identical at the amino acid level between Plasmodium knowlesi, Plasmodium yoelii, and P. falciparum. In summary, we describe a novel stage-specifically expressed, highly conserved gene product of P. falciparum that is recognized by parasite growth inhibitory antibodies.     

华南地区质粒介导超广谱β-内酰胺酶的基因分型研究

目的：了解华南地区质粒介导超广谱β－内酰胺酶（ESBLs）的发生率及基因型特征。方法：收集2001年4月－9月革兰阴性菌临床分离无重复株共1184株，采用NCCLS表型筛选和确认试验进行了ESBLs产酶的识别，E-test法检测各亚型ESBLs的MICs值，质粒接合及电转化实验，耐药质粒提取及酶切指纹分析，等电聚焦电泳，PCR通用引物扩增TEM、SHV、CTX－M、VEB、PER、SFO基因及其克隆测序进行ESBLs基因分型和质粒定位。结果：革兰阴性苗ESBLs的检出率为14．6％（173／1184）；获得产ESBLs接合子67株，电转化子11株，其中产CTX－M－14型ESBLs为33．3％（26／78）、CTX－M－3为23．1％（18／78）、CTX－M－9为14．1％（11／78）及SHV-2a为2．6％（2／78），未定型为5．1％（4／78）；29．5％（23／78）野生株伴广谱酶TEM－1或SHV－1型；各型ESBLs基因约定位在35－190kb大小的可接合性低执行者拷贝数天然质粒上；CTX－M型ESBLs以对头孢噻肟高水平高耐为特征。结论：华南地区质粒介导的ESBLs以CTX－M型衍生酶为主，其次是SHV型酶。     

Mild atopic dermatitis lacks systemic inflammation and shows reduced nonlesional skin abnormalities

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