吖啶橙染色法在膀胱癌无创诊断中的应用

[目的]运用吖啶橙染色荧光显微镜(acridineorenge,AO-F)检查法进行膀胱癌无创性诊断。[方法]采用AO-F检查法及巴氏染色分级(papanicolaousstaingradingsmethod,PG)检查法检测79例膀胱癌患者及80例非肿瘤患者的尿脱落细胞。[结果]两种染色法对诊断膀胱癌均有统计学意义,分层卡方检验,χMH2=173.461,P=0.000。巴氏染色法与吖啶橙荧光染色法比较,后者膀胱癌的检出率优于前者,χ2=19.606,P=0.000。巴氏染色法检出膀胱癌的敏感度为56.6%,特异度为100%。吖啶橙荧光染色法检出膀胱癌的敏感度为85.5%,特异度为100%。80例非膀胱癌患者两种尿脱落细胞检查的阳性检出率为0%。[结论]吖啶橙染色法比单从细胞形态上检测肿瘤细胞更具有重要的临床价值,可作为膀胱癌的辅助诊断方法。     

High-plasticity mesenchymal stem cells isolated from adult-retained primary teeth and autogenous adult tooth pulp—A potential source for regenerative therapies?

PurposeThe objective of this study was to compare the growth rate, morphology, immunohistology and plasticity of autogenous adult-retained SHEDs (arSHEDs) and adult dental pulp stem cells (DPSCs) obtained from the same donor.MethodsExpression of the mesenchymal stem cell markers CD44, CD90, CD105, caspase-3 and GAPDH were assessed using RT-PCR. Caspase-3 and CD44 were also evaluated at the protein level by western blotting of cell lysates. Plasticity of DPSCs and arSHEDs were tested by culture in adipogenic, chondrogenic, osteogenic and Schwann cells induction media.ResultsDPSCs and arSHEDs were isolated by explant culturing and were similarly positive for growth rate and all tested markers. Furthermore, DPSCs and arSHEDs could be driven to adipocyte, chondrocyte, osteocyte and Schwann cells lineages thus indicating similar plasticity as precursor cells.ConclusionThis study demonstrates the similarities between DPSCs and arSHEDs in a unique situation, where both stem cells (SC) types were obtained from a single patient and thus represent an alternative source of SC’s for tissue engineering and regeneration.     

结肠灌洗脱落细胞端粒酶活性诊断结肠癌的价值

目的端粒酶作为核糖核酸蛋白酶能使端粒延长而克服细胞衰老，与细胞永生和癌变密切相关，正常的躯体细胞无端粒酶活性，大多人类原发肿瘤均可探及端粒酶阳性．本研究旨在探讨结肠脱落细胞端粒酶检测对结肠癌的临床诊断价值．方法采用端粒重复序列扩增法（ＴＲＡＰ）检测１６例结肠癌外科手术新鲜标本、相邻的正常组织及其肠腔灌洗物，６例ＩＢＳ患者的结肠镜抽取物的端粒酶活性．结果结肠癌组织１６例，端粒酶强阳性１０例，弱阳性３例和阴性３例，阳性率为８１３％；结肠癌远端正常肠粘膜组织的端粒酶强阳性、弱阳性和阴性分别为０，２和１４例，阳性率为１２５％，两组Ｐ＜００１．结肠癌结肠灌洗液１６例中有５例端粒酶阳性，而６例ＩＢＳ结肠镜抽取液全部为端粒酶阴性，其诊断的敏感性和特异性分别为３１３％和１００％，阳性预测值和阴性预测值分别为１００％和６４７％．结论结肠灌洗脱落细胞的端粒酶检测对结肠癌有良好的诊断价值．     

口腔脱落细胞检测无精子因子微缺失的临床应用

目的 探讨同一个体口腔脱落细胞和血液检测无精子因子（AZF）缺失的相关性,研究口腔脱落细胞检测AZF 的可行 性。方法 选择287例研究对象,其中严重少弱精症120 例,无精症80例,正常男性87例。同一受试对象的血液和口腔脱落细胞样本统一编号。应用多重PCR 技术,分别检测口腔脱落细胞、血液基因组中AZF区域的Y 染色体微缺失情况。结果 血液和口腔脱落细胞检出AZF缺失的样本编号一一对应,并且缺失类型一致。在200 例患者中,29 例检出基因组AZF 基因微缺失,余171 例患者和87例正常对照者未检出AZF 基因微缺失。结论 在进行Y染色体微缺失筛查中,口腔脱落细胞检测AZF 简便、无创,可以替代血液AZF检测。     

阴道微环境和IL-10水平变化与高危型HPV感染的关系

目的 探讨阴道微环境和白介素-10(IL-10)水平变化与高危型HPV感染的关系。方法 选择2016年1月～2019年1月在舟山医院和甘肃省第二人民医院进行宫颈癌前筛查的患者3879例的资料进行回顾性分析,按照患者是否出现高危型HPV感染进行分组,分为病例组1346例,对照组2533例,通过单因素分析与多因素分析的方式,探讨阴道微环境和IL-10水平变化与高危型HPV感染的关系。结果 病例组患者的pH值、乳酸杆菌的量、白细胞级别与对照组比较差异均有统计学意义(t=50.598,χ2=244.382、27.604,P0.05),病例组患者的念珠菌检出率高于对照组,差异有统计学意义(χ2=284.363,P0.05),两组患者的滴虫检出率数据差异无统计学意义(χ2=2.802,P0.05),病例组患者阴道灌洗液中的IL-10浓度高于对照组患者,差异有统计学意义(t=62.260,P0.05),pH值偏高、乳酸杆菌的量偏少、白细胞偏多、念珠菌阳性、阴道灌洗液中的IL-10浓度偏高,均成为患者出现高危HPV感染的独立危险因素(P0.05)。结论 阴道微环境和IL-10水平变化与高危型HPV感染之间的关系密切,但需要更多研究对各类因素之间的因果关系进行深入分析。     

L1 C末端共同序列多肽诱导单克隆抗体应用于宫颈脱落细胞HPV的检出

目的 确定由人乳头瘤病毒( human papillomavirus,HPV)主要外壳蛋白(L1)C-末端保守序列多肽诱导的单克隆抗体对宫颈脱落细胞内的 HPV感染 是否具有可应用于临床的检测能力。方法 收集妇科门诊宫颈脱落细胞标本,一式两份,其中一份标本用于HPV感染的医院检验科基于市售HPV分型检测试剂盒检测,另一份以本课题组制备的HPV L1 C-末端保守序列多肽诱导的小鼠单克隆抗体作为探针用免疫组化(Immunological Histological Chemistry, IHC)方法检测标本中的 HPV L1,比较HPV L1 C-末端共同序列诱导单克隆抗体对宫颈脱落中 HPV 的检出结果和临床检测结果的差异。结果 收集到的 384 例妇科门诊宫颈脱落细胞标本中, HPV分型检测试剂盒检测出HPV阳性感染例数为63例,阳性率为16.41%(63/384);用免疫组化检测方法检测了其中380例标本,共检出阳性标本120例,阳性检出率为31.58%(120/380),与HPV分型检测试剂盒检测结果相比差异有统计学意义(P<0.05)。结论 初步证明了HPV L1 C- 末端保守序列多肽诱导的小鼠单克隆抗体对宫颈脱落细胞内 HPV 的感染具有良好的检出能力,并且检测能力明显要高于现临床所用的市售 HPV 分型检测试剂盒的检测能力(P<0.05)。该单克隆抗体在开发用于宫颈癌预防性筛查的 HPV 检测试剂盒方面具有一定的潜力。     

In vitro and in vivo characteristics of stem cells from human exfoliated deciduous teeth obtained by enzymatic disaggregation and outgrowth

Objective

Stem cells from human exfoliated deciduous teeth (SHED) are a good source of dental tissue for regeneration therapy, and can be obtained using different primary culture methods. The aim of this study was to determine the differences in the in vitro and in vivo characteristics between SHED isolated via enzymatic disaggregation (e-SHED) and outgrowth (o-SHED) primary culture methods.

Design

Dental pulp stem cells were isolated from 14 exfoliated deciduous teeth by enzymatic disaggregation (n = 7) and outgrowth (n = 7). Their proliferation potential and colony-forming ability were evaluated in vitro, as was their mesenchymal stem-cell-marker expression (using flow cytometry), and their differentiation was verified using quantitative real-time PCR (qPCR) and histochemical staining. In addition, the qualitative and quantitative characteristics of the hard tissue that was generated after in vivo transplantation were compared using haematoxylin and eosin staining, immunohistochemical staining, qPCR, and quantitative alkaline phosphatase analysis.

Results

The cell-proliferation potential, colony-forming ability, and Stro-1 and CD146 expression were higher in e-SHED than in o-SHED. While the in vitro adipogenic differentiation potential was greater in e-SHED than in o-SHED, the in vitro osteogenic differentiation did not differ significantly between the two cell types. Although in vivo hard tissue formation was greater following transplantation of o-SHED into mice, there was no difference in the quality of hard tissue generated by e-SHED and o-SHED.

Conclusion

The findings of this study indicate that e-SHED exhibit stronger stemness characteristics, but that o-SHED are more suitable for hard-tissue regeneration therapy in teeth.     

改良富血小板血浆促进乳牙牙髓干细胞成骨分化作用研究

目的：体外研究改良富血小板血浆（modified platelet-rich plasma,mPRP）促进人乳牙牙髓干细胞成骨分化的作用。方法：以α-MEM作为基础培养基,分别加入1％、2％、5％、10％4种不同浓度 mPRP 或者10％胎牛血清（对照）,对第4代SHED 连续培养并诱导矿化,碱性磷酸酶试剂盒检测 ALP 活性的变化,qRT-PCR 方法检测细胞内 RUNX2和骨钙素 mRNA 含量的改变。结果：不同浓度的 mPRP 均可以促进乳牙牙髓干细胞的 ALP 活性,且浓度为2％时 A 值最高;qRT-PCR 检测显示2％ mPRP 可以上调乳牙牙髓干细胞内 RUNX2及骨钙素 mRNA 的含量。结论：一定浓度的 mPRP 对乳牙牙髓干细胞的成骨分化具有一定的促进作用。