ABSTRACT
@#[摘 要] 目的：采用生物信息学方法探索与肾透明细胞癌(ccRCC)组织中铁死亡相关的lncRNA，并探讨其与免疫细胞浸润及患者预后的相关性，为ccRCC患者提供新的分子靶点。方法：从癌症基因组图谱(TCGA)数据库下载ccRCC的转录本数据和临床数据，利用单样本基因集富集分析(ssGSEA)及相关性分析获得与铁死亡相关的lncRNA；通过单因素和多因素回归分析构建与铁死亡相关的lncRNA特征图，分析其与预后的关系；利用R软件分析铁死亡相关lncRNA与肿瘤免疫细胞浸润和药物敏感性之间的关系。构建铁死亡相关RNA网络，并通过qPCR验证中国人ccRCC组织和癌旁组织（取自2019年12月至2021年03月间在西南医科大学附属医院手术切除8例标本）中关键lncRNA的表达。结果：Kaplan-Meier分析表明，铁死亡评分高的患者总OS率低于铁死亡评分低的患者。单因素和多因素回归分析确定11个ccRCC铁死亡相关lncRNA可评估患者预后，并构建ccRCC患者1、3、5年预后预测列线图。免疫细胞浸润分析表明，铁死亡相关lncRNA与ccRCC免疫细胞浸润密切相关，其中LINC01871、PRKAR1B-AS1和CYTOR是调节肿瘤免疫细胞浸润的关键lncRNA。化疗药物敏感性分析表明，高风险患者对甲氨蝶呤、紫杉醇、顺铂和多柔比星更为敏感。构建的包含3个lncRNA、15个miRNA和15个mRNA的RNA网络中，验证实验显示LINC01871、LINC00472和CYTOR在ccRCC组织中显著上调。结论：通过生物信息学方法获得11个与铁死亡相关的lncRNA，证明其与ccRCC组织免疫细胞浸润、化疗药物敏感性和患者预后相关，为探索ccRCC铁死亡相关lncRNA标志物提供重要参考。
ABSTRACT
Objective @#To investigate the inductive effects of canine periodontal ligament stem cells (PDLSCs) cocultured with canine disparate differentiating-period iliac bone marrow stromal cells (I-BMSCs).@*Methods@#Purified PDLSCs were isolated by in vitro culture of cBMSCs and flow cytometry. Third-generation PDLSCs were obtained, and conditioned culture medium derived fromiliac bone marrow stromal cells (I-BMSCs-CM) was added as indicated for coculture of PDLSCs. As the control group, pure uninduced PDLSCs were routinely cultured in DMEM culture medium containing 15%FBS. The experimental groups included the I-BMSCs-CM, I-BMSCs-CM-10ds and I-BMSCs-CM-15ds groups. The I-BMSCs-CM group consisted of PDLSCs induced by I-BMSCs, the I-BMSCs-CM-10ds group consisted of PDLSCs induced by I-BMSCs after osteogenic induction for 10 days, and the I-BMSCs-CM-15ds group consisted of PDLSCs induced by I-BMSCs after osteogenic induction for 15 days. The cocultured PDLSCs were examined via the MTT assay. Total mRNA and protein were prepared at 3 and 7 days. The mRNA expression levels of runt-related transcription factor 2(Runx2), special AT-rich sequence binding protein 2(Satb2) and osteocalcin (OCN) were measured by qRT-PCR. The protein expression levels of Satb2, Runx2 and OCN were detected by Western blot.@*Results@#The PDLSCs showed a spindle-like morphology. While the BMSC-conditioned media increased PDLSCs proliferation, the media conditioned by BMSCs allowed to differentiate for 15 days (I-BMSCs-CM-15days) significantly enhanced PDLSCs proliferation (F=342.8, P=0.017). The expression levels of the analyzed genes were upregulated in the coculture groups, and the protein expression levels of Satb2, Runx2 and OCN were higher in the test groups than in the control group at 7 days. At the protein level, I-BMSCs-CM-15days upregulated the expression of Satb2 by 3.04-fold (FSatb2=24.48, P=0.014), Runx2 by 5.1-fold (FRunx2=12.25, P < 0.001), and OCN by 3.67-fold (FOCN=18.35, P=0.022).@*Conclusion @#The conditioned medium of I-BMSCs may enhance the proliferation of PDLSCs, and that of terminally differentiated bone cells probably triggered the osteogenesis of PDLSCs, suggesting important implications for periodontal engineering.
ABSTRACT
Since the first successful pregnancy from a frozen human oocyte was reported, remarkable technological progress has been made in the area of cryopreservation of human oocytes. We report a successful delivery of two healthy babies after transfer of vitrified-warmed embryos derived from intracytoplasmic sperm injection (ICSI) with vitrified-warmed oocytes and frozen-thawed sperm. A female patient and her husband with severe oligoasthenspermia are reported. At the day of oocyte collection, very few inactive sperms were found in her husband semen. Multiple site open testicular biopsy was performed on her husband, but no sperm was retrieved. The patient did not become pregnant after transferring two embryos coming from half of oocytes and inactive sperms. The patient got pregnant and delivered two healthy babies after receiving a transfer of vitrified-warmed embryos from vitrified-warmed oocytes and frozen-thawed sperm.
La criopreservación de oocitos humanos ha progresado mucho desde que el primer embarazo exitoso desde un oocito congelado fue informado. Nosotros informamos el parto de dos bebés sanos después de transferir embriones vitrificados y recalentados y espermios descongelados. Se trata de una mujer y su marido con una oligoastenoespermia severa. En el día de la recolección de oocitos, se encontraron muy pocos espermios inactivos en el semen del marido. Se tomaron biopsias testiculares pero se encontraron muy pocos espermios inactivos. La mujer logró quedar embarazada y dio luz a dos bebés sanos después de recibir una trasferencia de embriones vitrificados y recalentados, y de espermios descongelados.
Subject(s)
Humans , Female , Pregnancy , Infant, Newborn , Adult , Oocytes , Pregnancy Outcome , Cryopreservation , Sperm Injections, Intracytoplasmic , Embryo Transfer , Pregnancy, TwinABSTRACT
Objective:ASCL-1 gene expression is closely related to pulmonary neuroendocrine (NE) tumors, such as small cell carcino-ma, atypical carcinoid, carcinoid, and large cell NE carcinoma. This study aimed to analyze ASCL-1 protein expression in lung adenocar-cinoma (AD) and its correlation with prognosis. Methods:ASCL-1 protein expression in 283 cases of AD was determined through immu-nohistochemical analysis and compared with the expression of traditional NE markers, including chromogranin A, CD56, and synapto-physin. Western blot was performed to detect the ASCL-1 protein expression levels in AD. Single factor Chi-square test and Logistic multiple factor regression analysis were conducted to explore the factors related to ASCL-1 expression. Kaplan-Meier survival analysis was applied to evaluate the prognosis of patients with AD. Results:Immunohistochemical analysis showed that ASCL-1 expression was positive in 48/283 (16.9%) AD and positively correlated with NE markers (0<r<1, P<0.05). Western blot revealed that ASCL-1 protein was expressed in 63 cases of AD, in their tumor-adjacent normal samples, and in 9 cases of AD. By contrast, this protein was not ex-pressed in tumor-adjacent normal tissues. Single factor Chi-square test and multivariate logistic regression analysis indicated that AS-CL-1 protein expression was associated with smoking, tumor differentiation, TNM staging, and lymph node metastasis. Kaplan-Meier survival analysis demonstrated that overall survival (OS) time was shorter in the ASCL-1-positive group than in the ASCL-1-negative group (P<0.05). Conclusion:ASCL-1 protein expression may serve as an independent prognostic factor for patients with lung AD.