Development, migration, and survival of mast cells

Mast cells play a pivotal role in immediate hypersensitivity and chronic allergic reactions that can contribute to asthma, atopic dermatitis, and other allergic diseases. Because mast cell numbers are increased at sites of inflammation in allergic diseases, pharmacologic intervention into the proliferation, migration, and survival (or apoptosis) of mast cells could be a promising strategy for the management of allergic diseases. Mast cells differentiate from multipotent hematopoietic progenitors in the bone marrow. Stem cell factor (SCF) is a major chemotactic factor for mast cells and their progenitors. SCF also elicits cell-cell and cell-substratum adhesion, facilitates the proliferation, and sustains the survival, differentiation, and maturation, of mast cells. Therefore, many aspects of mast cell biology can be understood as interactions of mast cells and their precursors with SCF and factors that modulate their responses to SCF and its signaling pathways. Numerous factors known to have such a capacity include cytokines that are secreted from activated T cells and other immune cells including mast cells themselves. Recent studies also demonstrated that monomeric IgE binding to FcωRI can enhance mast-cell survival. In this review we discuss the factors that regulate mast cell development, migration, and survival.     

High-resolution genotyping of 58 STRs in 635 Northern Han Chinese with MiSeq FGx ® Forensic Genomics System

Sequence polymorphisms were characterized at 27 autosomal STRs (A-STRs), 7 X chromosomal STRs (X-STRs), and 24 Y chromosomal STRs (Y-STRs) in 635 Northern Han Chinese with the ForenSeq DNA Signature Prep Kit on the MiSeq FGx Forensic Genomics System. Since repeat region (RR) and flanking region (FR) variation can be detected by massively parallel sequencing (MPS), the increase in the number of unique alleles and the average of gene diversity was 78.18% and 3.51% between sequence and length, respectively. A total of 74 novel RR variants were identified at 33 STRs compared with STRSeq and previous studies, and 13 FR variants (rs1770275883, rs2053373277, rs2082557941, rs1925525766, rs1926380862, rs1569322793, rs2051848492, rs2051848696, rs2016239814, rs2053269960, rs2044518192, rs2044536444, and rs2089968964) were first submitted to dbSNP. Also, 99.94% of alleles were concordant between the ForenSeq DNA Signature Prep Kit and commercial CE kits. Discordance resulted from the low performance at D22S1045 and occasionally at DYS392, flanking region deletions at D7S820 and DXS10074, and the strict alignment algorithm at DXS7132. Null alleles at DYS505 and DYS448 and multialleles at DYS387S1a/b, DYS385a/b, DYS448, DYS505, DXS7132, and HPRTB were validated with other MPS and CE kits. Thus, a high-resolution sequence-based (SB) and length-based (LB) allele frequencies dataset from Northern Han Chinese has been established already. As expected, forensic parameters increased significantly on combined power of discrimination (PD) and combined power of exclusion (PE) at A-STRs, mildly on combined PD and combined mean exclusion chance (MEC) at X-STRs, and barely on discrimination capacity (DC) at Y-STRs. Additionally, MiSeq FGx quality metrics and MPS performance were evaluated in this study, which presented the high-quality of the dataset at 20 consecutive runs, such as ≥ 60% bases with a quality score of 20 or higher (%≥ Q20), > 60% of effective reads, > 2000 × of depth of coverage (DoC), ≥ 60% of allele coverage ratio (ACR) or heterozygote balance, ≥ 70% of inter-locus balance, and ≤ 0.4 of the absolute value of observed minus expected heterozygosity (|H_exp – H_obs|). In conclusion, MiSeq FGx can help us generate a high-resolution and high-quality dataset for human identification and population genetic studies.     

A case report on concurrent occurrence of systemic mastocytosis and myeloid sarcoma presenting with extensive skin involvements and the results of genetic study

Introduction:Systemic mastocytosis is a rare disease due to mast cell accumulation in various extracutaneous sites. Systemic mastocytosis with an associated clonal hematologic non-MC lineage disease is the second most common subtype of systemic mastocytosis. The most common mutation associated with both systemic mastocytosis and myeloid sarcoma is mutation in Kit. Here, we identified the novel KIT D816V and ARID1A G1254S mutations co-occurring in systemic mastocytosis with myeloid sarcoma.Patient Concerns:A 33-year old male patient presented multiple skin lesions for 10 years. Symptoms accelerated in 2017 with decreased body weight. Physical examination revealed enlarged lymph nodes in his neck, axilla and inguinal region; conjunctival hemorrhage; gingival hyperplasia. Skin biopsy showed mast cell infiltration. Flow cytometry detected CD2, CD25 and CD117 positive cells in lymph nodes. Codon 816 KIT mutation D816V and codon 1245 ARID1A mutation G1254S were found in peripheral blood. MPO, CD117, CD68 positive cells in lymph nodes indicated co-existing myeloid sarcoma.Diagnosis:Systemic mastocytosis with an associated clonal hematologic non-MC lineage disease of myeloid sarcomaInterventions:Cytarabine and daunorubicin for myeloid sarcoma and dasatinib for systemic mastocytosis were initiated. Anti-histamine and anti-leukotrienes therapy were used to prevent NSAIDs-induced shock. Platelets were infused to treat bone marrow suppression.Outcomes:Patient was discharged after recovered from bone marrow suppression. Dasatinib continued on outpatient.Conclusion:This is the first case of patient with systemic mastocytosis and myeloid sarcoma simultaneously presenting extensive skin involvements. Mutations of Kit and Arid1a emphasis the importance to notice possibility of various tumors occurring in patients with multiple mutations. In addition, cysteine-leukotrienes-receptor antagonists should always be used to prevent anaphylactic shock due to mast cell activation.     

两种转染人宫颈癌HeLa细胞的方法学比较

目的比较两种不同的转染方法,即Lipofectamine2000和Nucleofector Kit转染人宫颈癌Hela细胞的转染效率。方法分别选取Lipofectamine2000转染和Amax核电转(Amax Nucleofector Kit)两种转染方法。HeLa细胞培养贴壁达到85%~90%后传代获取HeLa单细胞悬液,细胞分成2管,其中1管直接采用核电转法转染红色荧光蛋白(DsRed)后种植于1.5 ml的细胞培养皿;另一管先种植于细胞培养皿,待贴壁24 h后采用Lipofectamine2000转染DsRed。细胞的存活率采用台盼兰进行检测。分别比较两者转染效率及转染前后细胞的存活率,进一步探讨Nucleofector Kit核电转法的高效性。结果 Lipofectamine2000的基因转染率仅为20.23%,而核电转法的转染率为90.14%,后者为前者的4.46倍。Lipofectamine2000转染细胞前后的存活率分别为96.34%和90.76%,而核电转组分别为95.98%和78.35%。结论细胞核电转法能高效稳定地转染人宫颈癌细胞株HeLa细胞。     

国内外16种弓形虫诊断试剂盒的评估

目的评估市售16种弓形虫抗体诊断试剂盒。方法采用敏感性、特异性、总一致性（crude agreement）及约登指数（Youden index）等分析指标作为评判标准,比较16种试剂盒对弓形虫IgG或IgM阳性血清和正常人血清的检测效果。结果4种进口弓形虫IgG试剂盒中,3种试剂盒的敏感性和特异性高于97．0％,4种国产同类试剂盒中,2种试剂盒的敏感性和特异性大于80．0％,这5种试剂盒约登指数均大于0．8。4种进口弓形虫IgM试剂盒中,2种试剂盒的敏感性和特异性高于87．0％,国产试剂盒仅1种敏感性为87．5％,其特异性仅为31．1％,其余3种敏感性低于6．25％;约登指数仅2种进口试剂高于0．8。结论进口试剂盒的检测质量优于国产试剂盒,国内研制的市售弓形试剂盒的检测效果和质量存在一定的问题,尤以IgM试剂盒为甚。     

日本血吸虫病间接血凝试验诊断试剂盒临床诊断效能

目的评价日本血吸虫病间接血凝试验（IHA）诊断试剂盒对血吸虫病的临床诊断效能。方法分别在江西省、安徽省和湖北省3个疫区现场进行血吸虫病调查,并采集371份病原学确诊的血吸虫病人血清（毛蚴孵化法或/和Kato-Katz法阳性,阳性金标准）,761份无血吸虫感染史、无疫水接触史且病原学检测阴性者血清（阴性金标准）。采用日本血吸虫病IHA诊断试剂盒对上述血清进行血吸虫抗体检测,并计算阳性符合率（灵敏度）、阴性符合率（特异度）、阳性预测值和阴性预测值等诊断效能指标。结果日本血吸虫病IHA诊断试剂盒与阳性金标准的符合率为95.1%（95%可信区间：92.9%～97.3%）,阴性金标准的符合率为97.8%（95%可信区间：96.8%～98.8%）,阳性预测值和阴性预测值分别为95.4%和97.6%。试剂盒对不同感染度的阳性金标准血清（1～、41～、100～）检出的抗体阳性率之间差异无统计学意义;各年龄组间IHA灵敏度和特异度变化较小,差异均无统计学意义。结论日本血吸虫病IHA诊断试剂盒对血吸虫病具有较好的诊断效能,可用于血吸虫病的辅助诊断,适用于疫区大规模人群的血吸虫病现场筛查。     

金标免疫渗滤法诊断试剂盒诊断血吸虫病的价值研究

目的　评价金标免疫渗滤法诊断试剂盒（DIGFA-kit）用于血吸虫病血清流行病学调查的价值。方法　采用DIGFA与ELISA单盲平行检测各期血吸虫病病人血清、非流行区人群血清、疫区人群血清和血吸虫病已控制地区人群血清共2304份。结果　DIGFA检测血吸虫抗体的敏感性为96.8%,对非流行区健康人群的特异性为100%;该法检测疫区居民血清508份,抗体阳性检出率为35.6%,在Kato法粪检阳性的81人中,其中抗体阳性79人,阳性符合率为97.5%;检测血吸虫病已控制地区人群血清,抗体阳性率为4.6%;根据DIGFA的敏感性和对非流行区人群的特异性,计算其Youdens指数为0.926。上述各项结果与ELISA比较,差异无显著性（P>0.05）。结论　DIGFA与ELISA有相似的敏感性和特异性,且方法简便、快速,胶体金标记物制备容易,试剂稳定,运送方便,在现场和临床诊断更具有优越性。     

长爪沙鼠胃肠道Cajal间质细胞的形态学

目的 探讨长爪沙鼠胃肠道Cajal间质细胞（ICCs）的形态和分布规律。 方法 采用10只成年长爪沙鼠,体重50~70g,取胃、小肠、结肠制作冷冻切片,结合全层铺片的c-Kit免疫荧光染色。结果 ICCs呈网络状分布于整个胃肠道,不同部位ICCs的分布及形态有所不同。在胃底部,仅见肌内ICCs(ICC-IM）,而在胃体和胃窦部除ICC-IM外,可见肌间ICCs(ICC-MY)分布在肌间神经丛周围;其细胞密度胃底ICC-IM最多,由胃底至胃窦逐渐减少,而ICC-MY由胃体至胃窦逐渐增多。在小肠可见ICC-IM, ICC-MY和深肌层ICCs(ICC-DMP)3个亚群,结肠管壁内也分布有ICC-IM、ICC-MY和黏膜下ICCs(ICC-SM)3个亚群。结论 沙鼠可用于有关ICCs正常形态、结构及功能的研究。     

Analytical evaluation of hemoglobin A(1c) dual kit assay on Bio-Rad Variant II: an automated HPLC hemoglobin analyzer for the management of diabetic patients

OBJECTIVES: To evaluate the analytical performance of the Bio-Rad Variant II HbA(1c) dual kit assay. DESIGN AND METHODS: Precision, carryover, linearity and analytical range were investigated. 139 patients' HbA(1c) results analyzed by the Variant II were compared to the Variant I method. 49 blood samples analyzed by the Variant II at Toronto Medical Laboratories (TML) were compared to the Variant II at Hospital for Sick Children (HSC). RESULTS: Total imprecision was less than 2% for the Variant II assay. The method had a wide analytical range with no carryover. HbA(1c) results were not changed after switching back and forth from the beta thalassemia to HbA(1c) assay. The Variant II showed an average of 0.0027 negative bias compared to the Variant I method. There was an average of 0.0020 negative bias for HbA(1c) results on the Variant II at TML compared to the Variant II at HSC. CONCLUSIONS: HbA(1c) analysis on the Variant II HbA(1c) dual kit is a relatively fast and reproducible method.