小鼠趋化因子COL19Ig融合蛋白表达载体的构建、表达及鉴定

通过特异引物扩增出去掉终止密码的mCCL19编码序列，经酶切、亚克隆、拼接构建了真核表达载体pcDNA3．1-CCL19Ig，酶切和测序鉴定插入序列；将重组质粒转染CHO细胞进行体外表达，通过RT—PCR、Western blot鉴定目的基因的表达，利用趋化小室法测趋化活性。结果测序证实重组表达载体含mCCL19编码序列和人IgGI Fc段序列，其序列分别与GenBank中公布序列比对一致，IgG1-Fc段读码框未发生改变；Western blot结果证实转染了pcDNA3．1-mCEL19Ig的CHO细胞培养上清中有相对分子质量为38000的融合蛋白mCCL19Ig表达；体外趋化实验表明融合蛋白mCCL19Ig对小鼠脾细胞有趋化活性，且呈剂量依耐关系。     

Nestin、CK19及insulin等在胚胎胰腺发育中的表达

目的:研究Nestin、CK19、胰岛素、胰高血糖素及生长抑素在胚胎胰腺发育中的表达,探讨胰岛细胞分化发育的机制。方法:采用免疫组化SABC法及免疫组化双染法(LAB SA),对30例6～14wk人胚胎胰腺中,Nestin、CK19、胰岛素,胰高血糖素及生长抑素阳性的细胞进行定位。结果:(1)胚胎胰腺发育中Nestin阳性细胞存在于胰腺的间质,数量极少;CK19在胰腺导管上皮分化中持续表达,呈强阳性;(2)7wk胰腺导管上皮开始分化出胰岛细胞,胰岛素、胰高血糖素及生长抑素表达阳性,三者的表达并无时段差异性;随着胎龄的增加,阳性细胞数增加,14wk时胰岛逐渐形成,表达达到高峰。结论:胚胎胰腺的发育中,胰腺间质存在胰腺干细胞;胚胎早期胰腺导管上皮细胞开始分化并分泌胰岛素,其自分泌的激素可能参与调节胰岛细胞的迁移和胰岛的形成。     

Detection of single nucleotide substitution by competitive allele-specific short oligonucleotide hybridization (CASSOH) with immunochromatographic strip

Recent advances in human genome research have revealed that genetic polymorphisms, such as single nucleotide polymorphisms (SNPs), are closely associated with susceptibility to various common diseases and adverse drug reactions. Also, numerous mutations responsible for a number of genetic diseases have been identified. Clinical application of genetic information to individual health care requires simple and rapid identification of nucleotide changes in clinical settings. We have devised a novel low-tech method for the detection of a single nucleotide substitution using competitive allele-specific short oligonucleotide hybridization with immunochromatographic strip. The gene of interest is PCR-amplified, hybridized to an allele-specific short oligonucleotide probe in the presence of a competitive oligonucleotide, and subjected to chromatography using a DNA test strip at room temperature. The genotype is unambiguously determined by the presence or the absence of visible purple lines on a strip. Feasibility of the method was demonstrated by the detection of a prevalent disease-causing mutations in glycogen storage disease type Ia (G6PC), medium-chain acyl-CoA dehydrogenase deficiency (ACADM), non-ketotic hyperglycinemia (GLDC), and clinically important polymorphisms in the CYP2C19 gene and the aldehyde dehydrogenase 2 gene (ALDH2). The procedure does not demand either technical expertise or expensive instruments and is readily performed in local clinical laboratories. The result is obtained within 10 min after PCR. This rapid and simple method of SNP detection may be used for point-of-care genetic diagnosis with potentially diverse clinical applications. Hum Mutat 22:166-172, 2003.     

Immuno-proteomic profiling reveals aberrant immune cell regulation in the airways of individuals with ongoing post-COVID-19 respiratory disease

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Distinction between papillary thyroid hyperplasia and papillary thyroid carcinoma by immunohistochemical staining for cytokeratin 19, galectin-3, and HBME-1

The histopathology of papillary thyroid hyperplasia and papillary thyroid carcinoma is similar enough to cause a diagnostic dilemma in a few cases. Both lesions may have papillary fronds with fibrovascular cores, nuclear crowding, and nuclear anisocytosis. Formalin-fixed paraffin-embedded tissues from 30 randomly selected patients with papillary thyroid hyperplasia and an equal number from patients with papillary thyroid carcinoma were analyzed for expression of cytokeratin 19 (CK19), galectin-3, and HBME-1. Cases of papillary thyroid carcinoma had moderate to strong CK19, galectin-3, and HBME-1 reactivity although both CK19 and galectin-3 showed positive staining in a significant number of nonneoplastic thyroid cases. HBME-1 was uncommon in the nonneoplastic cases. These results indicate that HBME-1 may be useful in helping to distinguish papillary thyroid carcinoma from hyperplasia in diagnostically difficult cases.     

New genome type of adenovirus serotype 19 causing nosocomial infections of epidemic keratoconjunctivitis in Japan

Twelve strains of adenovirus serotype 19, isolated from cases of epidemic keratoconjunctivitis in Japan in 1992, 1993, 1997, and 1998, were analyzed by DNA restriction analysis, using restriction endonucleases BamHI, BglI, BglII, EcoRI, HindIII, KpnI, PstI, SacI, SalI, SmaI, and XhoI. Among these 11 restriction endonucleases, EcoRI, PstI, SacI, and SmaI were discriminative enzymes, showing restriction patterns different from those reported previously for the prototype and the variant 19a. This new genome type was isolated in 1997 and 1998, when an increase of epidemic keratoconjunctivitis cases caused by adenovirus serotype 19 was observed for both sporadic and nosocomial infections. Strains from 1992 and 1993 showed restriction patterns similar to those of the worldwide reported variant 19a for all enzymes used. The changes detected in strains from 1997 and 1998 could be the reason for the recent epidemic.     

Neurological Manifestations of COVID-19 Feature T Cell Exhaustion and Dedifferentiated Monocytes in Cerebrospinal Fluid

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人细小病毒B19检测方法研究进展

人类微小病毒B19（HPV B19）是细小病毒属中唯一对人类有致病性的病原体。感染后可引起传染性红斑、关节病、再生障碍性贫血危象、胎儿水肿及流产，对某些免疫力低下的病人，B19病毒感染甚至是致命的。B19病毒一般通过呼吸道传播，也可经输血和血液制品传播。建立简单实用的检测方法对B19病毒相关疾病的诊断和治疗非常重要。本文拟对B19病毒检测方法作一综述。