目的 分析芳香化酶(CYP19)基因多态性与子宫内膜异位症(endometriosis,EMs)术后复发的关系。方法 回顾性分析2019年2月—2020年2月于我院接受手术的110例EMs患者临床资料,按照术后12个月是否复发分为未复发组(62例)、复发组(48例),通过聚合酶链反应-限制性片段长度多态性(PCR-RFLP)技术测定两组CYP19基因115T/C、240A/G、1531C/T位点的单核苷酸多态性(single nucletide polymorphism,SNP),并分析不同位点对应VAS评分、r-AFS评分的差异。结果 复发组CYP19基因115T/C、1531C/T位点不同基因型及等位基因频率与未复发组相比,差异均无统计学意义(P＞0.05);复发组CYP19基因240A/G位点AG基因型频率比未复发组高(P＜0.05),AA基因型频率比未复发组低(P＜0.05);两组240A/G位点等位基因频率相比,差异有统计学意义(P＜0.05);CYP19基因240A/G位点AG 型VAS评分、r-AFS评分＞GG型＞AA型,差异有统计学意义(P＜0.05);CYP19基因115T/C、1531C/T位点不同基因型的VAS评分、r-AFS评分相比,差异均无统计学意义(P＞0.05)。结论 CYP19基因240A/G位点多态性与EMs术后复发、疼痛程度及病情密切相关,且携带G等位基因的基因型(AG+GG)可能是术后复发的风险因素。

Objective To analyze the relationship between aromatase (CYP19) gene polymorphism and recurrence of endometriosis (EMs) after surgery. Methods The clinical data of 110 patients with EMs who underwent the operation in our hospital from February 2019 to February 2020 were analyzed retrospectively. The patients were divided into non-relapsing group (62 cases) and relapsing group (48 cases) by 12 months followed-up outcomes. The single nucleotide polymorphism (SNP) of 115T/C, 240A/G and 1531C/T sites of CYP19 gene were detected by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP), and the differences of VAS and r-AFS scores between the two groups were analyzed. Results The genotypes and allele frequencies of 115T/C and 1531C/T of CYP19 gene in relapsing group were not significantly different from those in non-relapsing group (P>0.05). The AG genotype frequency of 240A/G site of CYP19 gene in relapsing group was higher than that in non-relapsing group (P<0.05), while the AA genotype frequency was significantly lower than that in non-relapsing group (P<0.05). There were significant differences in the allele frequencies of 240A/G site (P<0.05). The scores of VAS and r-AFS of AG＞GG＞AA, with significant differences (P<0.05). There were no significant differences in the VAS and r-AFS scores of CYP19 gene at 115T/C and 1531C/T site (P>0.05). Conclusion The 240A/G polymorphism of the CYP19 gene is closely related to postoperative recurrence, pain degree and condition of EMs, and the genotypes carrying the G allele (AG+GG) may be the risk factor of postoperative recurrence.

目的 对3例儿童Rotor综合征的临床特点及SLCO1B1和SLCO1B3基因突变分析,提高儿科医生对Rotor综合征的认识。方法 收集广州市妇女儿童医疗中心2018年—2019年确诊的3例Rotor综合征患儿的临床资料,对患儿及其家系成员肝脏常见遗传代谢性疾病二代测序筛查并家系验证结果进行分析。结果 患儿主要临床表现为反复或持续巩膜和(或)皮肤轻度黄染,实验室检查提示高直接胆红素血症。二代测序发现3例患儿均为SLCO1B1基因c.1738C＞T纯合突变和SLCO1B3基因5号内含子区域大片段插入纯合突变。SLCO1B1基因和SLCO1B3基因2处纯合突变均进行了家系验证。文献报道的SLCO1B1基因c.1738C＞T突变是无义突变,可以造成蛋白功能缺失;SLCO1B3基因的大片段插入突变虽暂未有文献收录或报道,但大片段的插入突变可引起移码突变而造成编码蛋白功能丧失。结论 由于基因检测技术的不断进步,Rotor综合征不断被儿科医生所认识。SLCO1B1和SLCO1B3双基因纯合或复合杂合突变是3例Rotor综合征患儿的分子遗传基础。

Objective To better understand Rotor syndrome(RS)in children,the clinical features and SLCO1B1 and SLC01B3 gene mutations were analyzed. Methods The clinical data of the 3 pediatric cases diagnosed in Guangzhou Women and Children's Medical Center between 2018 and 2019 was collected. Genomic DNA was extracted from the children and their family members, and subjected for second-generation sequencing to screen the known genes for liver genetic metabolic diseases. Then the detected mutations were confirmed by Sanger sequencing analysis. Results The main clinical manifestations were recurrent or persistent mild yellowish sclera and/or skin. Laboratory examinations showed hyperbilirubinemia with direct bilirubin elevating. Second generation sequencing showed that all 3 children were c.1738c＞Thomozygous mutations of SLCO1B1 gene and homozygous mutations of large fragment insertion in SLCO1B3 gene intron 5. Two homozygous mutations in SLCO1B1 gene and SLCO1B3 gene were verified in families.SLCO1B1 gene c.1738C＞T mutation,a nonsense mutation reported in references,could lead to protein function loss.A large insertion mutation of SLCO1B3 gene could cause frame-shift mutation which might lead to protein function loss even though it was neither reported in the references nor recorded in SNP database. Conclusion Due to the progress in the clinical application of gene detection technology, RS has been recognized gradually by pediatricians. Homozygous mutations or compound heterozygous mutations simultaneously occurred in SLCO1B1 and SLCO1B3 gene were the molecular genetics base in these cases of RS.

目的 探讨精神分裂症患者亚甲基四氢叶酸还原酶 (MTHFR)基因多态性、血浆HCY水平与心血管疾病发生风险的相关性。方法 收集住院精神分裂症患者164例,记录一般资料和测定患者MTHFR C677T基因多态性、血浆同型半胱氨酸(HCY)水平、总胆固醇(TC)、高密度脂蛋白胆固醇(HDL-C)、Framingham心血管疾病发生风险评分(FRS)等指标。其中10年心血管疾病发生风险用 Framingham心血管疾病风险评分表来计算。结果 1.CC、CT、TT基因型血浆HCY水平呈增高趋势,差异有统计学意义(P<0.001);CC基因型和CT基因型血浆HCY水平与FRS评分呈正相关关系(r_s=0.27,P=0.016;r_s=0.42,P=0.002);TT基因型血浆HCY水平与FRS评分之间无相关性(r_s=0.05,P=0.784)。2.低、中、高风险组的血浆HCY水平差异无统计学意义;低风险组和中风险组血浆HCY水平与FRS评分呈正相关关系(r_s=0.29,P=0.000;r_s=0.55,P=0.006);高风险组血浆HCY水平与FRS评分无相关性(r_s=-0.16,P=0.66)。结论 精神分裂症病人血浆HCY水平与FRS评分具有相关性,MTHFR C677T基因多态性、血浆HCY水平是对精神分裂症病人心血管疾病风险评估的合理补充之一。

目的 通过对43种融合基因在儿童白血病中的结果分析,探讨融合基因阳性的儿童急性B淋巴细胞白血病(acute B-lymphoblastic leukemia,B-ALL)的免疫表型特征。方法 应用实时荧光探针PCR法对2016年10月—2018年12月在深圳市儿童医院就诊的初发或复发B-ALL患儿进行融合基因检测,采用多参数流式细胞术(flow cytometry,FCM)对B-ALL患者进行免疫表型检测。结果 120例B-ALL患儿融合基因筛选总阳性率为37.5%(45/127),包括TEL/AML1 27例、E2 A/PBX1 7例、BCR/ABL1 6例、MLL 4例、TLS/ERG 1例;不同年龄段白血病融合基因阳性率差异有统计学意义(P<0.01),性别分布差异无统计学意义(P>0.05)。各融合基因阳性组CD19阳性率为100%,TEL/AML1阳性表达患者普通-B-ALL表型占比最高(77.8%),干/祖细胞抗原CD34的阳性率为81.5%;E2 A/PBX1阳性表达患者以前-B-ALL表型为主,不表达已知的T系及髓系抗原;各融合基因阳性组及阴性组患儿髓系抗原阳性率比较差异有统计学意义(P<0.01),以BCR/ABL1基因表达组阳性率最高(100%)。结论 5种融合基因在患者年龄构成及免疫表型中具有一定的分布特点;B-ALL特征性免疫表型的改变可用于融合基因表达的预测,提高融合基因结果判读的准确率。

Objective To investigate the immunophenotype features of children with acute B-lymphoblastic leukemia(B-ALL) combined with fusion gene expressing after to analyze the results of the 43 fusion genes. Methods Real-time fluorescent probe PCR assay was used for the detection of fusion genes in 120 cases of children from Shenzhen Children's Hospital with B-ALL newly or recurrently diagnosed from Oct 2016 to Dec 2018. Multi-parameter flow cytometry(FCM) was used for the detection of the immunophenotype in children with B-ALL. Results Of all the 120 cases, the fusion genes were detected at positive rate of 37.5%(45/120), included TEL/AML1 27 cases, E2 A/PBX1 7 cases, BCR/ABL1 6 cases, MLL 4 cases, TLS/ERG 1 cases. The positive rate of leukemia fusion gene had statistically difference among fusion genes positive groups based on age(P<0.01). There was no statistically difference in the gender distribution(P>0.05). The expressing of CD19 was at positive rate of 100% in all of the groups. The rate of the common-B-ALL was the highest B-ALL subtype in the TEL/AML1 positive groups(77.8%). The stem /progenitor associated antigen CD34 was at positive rate of 81.5%. The pre-B-ALL was the main subtype in the E2 A/PBX1 group, which was no expression of the known T-ALL associated antigen MyAg antigen. There was statistically difference in the positive rate of MyAg expression among all of the groups(P<0.01), with the highest rate in the BCR/ABL1 group(100%). Conclusion There were certain distribution features in age composition and immunophenotype of children with B-ALL carrying five kinds of common fusion genes. The characteristic changes of the immunophenotype of B-ALL may be used to predict the expression of fusion genes and improve the accuracy of fusion genes by the supplementary role of immunophenotype analysis.

目的 探讨具有不同载脂蛋白E4等位基因(Apolipoprotein E4 alleles,APOE4)阿尔茨海默病患者的神经心理学量表差异。方法 纳入2014年1月—2017年12月广州市第一人民医院收治阿尔茨海默病患者28人,分别予简易精神状态检查量表、阿尔茨海默病评定量表-认知部分、临床医师通过面谈对变化的印象、日常生活活动能力量表、神经精神问卷,并检测量表间相关关系。之后随访18个月,观察量表评测的各功能变化及互相间相关性。检测不同载脂蛋白E4等位基因等阿尔茨海默病相关基因分布及与量表间相关关系。结果 认知评定量表间、认知评定量表与整体评价量表间、以及认知评定量表与日常活动能力评定量表间具有相关性。精神与行为症状量表分数与其他评定量表无明显相关性。随访中各量表分数变化间均无相关性。各基因组间功能变化无显著性差异,载脂蛋白E4等位基因变异主要影响患者的认知功能。等位基因分布与患病年龄,日常活动能力及精神与行为症状无相关性。结论 阿尔茨海默病量表评测的各认知领域间相关性不同,功能变化间无相关关系。载脂蛋白E4等位基因变异主要影响患者的认知功能。

Objective The present study aimed to elucidate the performance of multiple psychological tests among different Apolipoprotein E4 alleles (APOE4) in people with Alzheimer's disease (AD). Methods 28 patients were enrolled from January 2014 to December 2017 in Guangzhou First People'S Hospital. All patients were tested by using Mini-mental State Examination (MMSE), Alzheimer's disease Assessment Scale (ADAS-cog), Clinician's Interview-Based Impression of Change (CIBIC-Plus), Activities of Daily Living (ADL) and the NeuroPsychiatric Inventory (NPI). After 18 months follow-up visit, the change of the tests points were recorded. AD pathogenic genes, including Apolipoprotein E4 allele's variations, were detected in all patients. Then the correlations of APOE4 alleles and multiple psychological tests were analyzed. Results The correlations were confirmed between MMSE and ADAS-cog, MMSE and CIBIC-plus, MMSE and ADL, ADAS-cog and CIBIC-plus, ADAS-cog and ADL. NPI showed no correlation with the others. No correlation was found between changes of multiple psychological tests after 18 months follow-up. APOE4 alleles' variation affected cognitive function mainly. The effects of APOE4 on ADL and NPI showed no statistical significance in AD patients. No correlation was found among patients groups with different APOE4 alleles in all psychological tests and age of onset. Conclusion The correlations were existed among multiple cognitive domains while levels were different. The changes between psychological tests showed no correlations. APOE4 alleles' variation affected cognitive function mainly.

目的 探讨河源地区机采血小板固定献血者血小板抗原系统的基因多态性特征,为建立本地区机采血小板供血者库奠定基础。方法 采用PCR-SSP方法对100例机采血小板固定献血者进行血小板抗原HPA1～17系统基因分型。结果 HPA1～17基因中成多态性分布的等位基因是HPA2a、HPA3a、HPA5a、HPA15a,其频率分别为0.96、0.49、0.99、0.515。HPA-2、HPA-3、HPA-5、HPA-15系统存在aa、ab、bb 三种表型。HPA1a、HPA4a、HPA6a-14a、HPA16a-17a基因频率为1,呈单线性分布,未发现b基因。结论 河源地区血小板HPA-3系统不配合率最高(0.420),HPA-15系统次之。建立本地区机采血小板供血者库,为患者提供HPA相合的血小板,对减少临床血小板输注无效的发生具有重要意义。

Objective To study the polymorphism of human platelet antigens in fixed apheresis platelet donors in Heyuan area and to lay a foundation for the establishment of platelets donor bank. Methods PCR-SSP method was used to analyze HPA 1～17 genotype in 100 fixed platelet donors. Results The highest numbers of heterozygotes were HPA2a,HPA3a,HPA5a and HPA15a,with frequencies of 0.96,0.49,0.99 and 0.515,respectively. The frequencies of HPA1a,HPA4a,HPA6a-14a and HPA16a-17a genes were 1,which showed a single linear distribution. Conclusion HPA-3 system were the highest mismatch rate (0.420),followed by HPA-15 system. It is great significance to establish a local platelet donor bank and provide HPA compatible platelets for patients.

目的 探讨精神分裂症患者载脂蛋白E基因多态性与血清ApoE浓度、血脂、心血管疾病发生风险的相关性。方法 收集住院精神分裂症患者116例,记录一般资料和测定患者载脂蛋白E基因(APOE)、载脂蛋白E(ApoE)、总胆固醇(TC)、高密度脂蛋白胆固醇(HDL-C)、心血管疾病发生风险评分(Framingham risk score,FRS)等指标。结果 ①APOEε2、ε3、ε4不同等位基因组精神分裂症患者TC、载脂蛋白E、FRS评分差异有统计学意义(P=0.01,P=0.005,P=0.012)。②载脂蛋白E与TC、FRS评分存在负相关关系(r_s=-0.48,P=0.02;r_s=-0.52,P=0.04),APOE ε4等位基因组载脂蛋白E与TC、FRS评分存在更高的相关关系(r_s=-0.55,P<0.001;r_s=-0.63,P=0.04)。结论 精神分裂症患者ApoE基因多态性与载脂蛋白E、胆固醇、FRS评分存在关联,ApoE基因-载脂蛋白E-胆固醇代谢通路可能是精神分裂症患者心血管疾病的致病机制之一。

目的 构建靶向CXCR7基因的CRISPR/Cas9基因编辑系统,并应用于HEK 293T细胞系。方法 设计两对靶向CXCR7基因的sgRNAs,分别插入PX458载体中,并转化DH5α大肠埃希菌。经菌液PCR和测序验证,挑选序列正确的sgRNA-CXCR7-PX458质粒,转染HEK 293T细胞,用流式分选转染阳性细胞,提取其DNA,PCR扩增后测序验证。结果 经测序验证,成功构建了靶向CXCR7基因的CRISPR/Cas9系统,转染HEK 293T细胞后,测序鉴定发现成功编辑CXCR7基因。结论 成功构建了靶向CXCR7的sgRNA-CXCR7-PX458质粒,可在HEK 293T上成功编辑CXCR7基因,为进一步的功能研究奠定基础。

Objective To construct the CRISPR/Cas9 gene editing system targeting C-X-C chemokine receptor 7 (CXCR7) gene and to edit CXCR7 gene in 293T cell line. Methods Two pairs of small guide RNAs (sgRNAs) targeting CXCR7 gene were designed and inserted into PX458 vector, which were transformed into host bacterium Escherichia coliDH5α. The correct sgRNA-CXCR7-PX458 plasmids were selected by PCR and further Sanger sequencing verification. HEK 293T cell line was transfected by DNA of sgRNA-CXCR7-PX458 plasmid. After 72 hours,GFP-positive cells were sorted by flow cytometry. We did DNA extraction of the GFP-positive cells and amplified the CXCR7 gene corresponding fragment by PCR and investigated the CXCR7 gene editing results by Sanger sequencing. Results The CRISPR/Cas9 system targeting CXCR7 gene was successfully constructed. After 293T cells were transfected, the CXCR7 gene was edited in HEK 293T cells successfully. Conclusion The sgRNA-CXCR7-PX458 plasmid targeting CXCR7 gene was successfully constructed. The CRISPR/Cas9 gene editing system targeting CXCR7 gene were used on the HEK 293T cell line, which lays a foundation for further study of BCOR function.

目的 分析人脑胶质瘤组织中O6—甲基鸟嘌呤—DNA甲基转移酶(MGMT)、DNA拓扑异构酶Ⅱ(TopoⅡ)基因的表达情况及其对化疗敏感性的影响。方法 收集医院2012年4月—2018年6月期间进行开颅手术切除的新鲜人脑胶质瘤标本80例和同期经颅脑手术治疗的脑外伤或脑出血内减压切除的正常脑组织30例。采用免疫组化法检测两种标本中MGMT和TopoⅡ基因表达情况,并通过脑胶质瘤U251和U87细胞培养、体外药物(替莫唑胺)干预、Transwell体外侵袭实验分析其对化疗敏感性的影响。结果 胶质瘤标本、正常脑组织MGMT和TopoⅡ基因表达程度分布比较差异均有统计学意义(P＜0.05),且二者MGMT基因阳性表达率分别为63.75%、3.33%,TopoⅡ基因阳性表达率分别为55.00%、0.00%,差异有统计学意义(P＜0.05)。MGMT、TopoⅡ基因均在细胞核显示为阳性染色。体外药物干预的实验组、未进行药物干预的阴性对照组干预前U251、U87细胞穿膜细胞计数比较无统计学意义(P＞0.05),但干预后实验组U251、U87细胞穿膜细胞计数高于阴性对照组(P＜0.05),干预后实验组U251、U87细胞有更强的侵袭力。结论 MGMT和TopoⅡ基因在人脑胶质瘤标本中阳性表达率高,且可能参与促进脑胶质瘤细胞侵袭,影响肿瘤化疗敏感性。

Objective To analyze the expression of O6-methylguanine-DNA methyltransferase (MGMT) and DNA topoisomerase II (Topo II) genes in human brain gliomas and its influence on the chemosensitivity. Methods A total of 80 fresh human brain glioma specimens resected by craniotomy and 30 normal brain tissues resected by craniocerebral operation for traumatic brain injury or decompression for cerebral hemorrhage during the period from April 2012 to June 2017 were collected. The expression of MGMT and Topo II genes in the two kinds of specimens was detected by immunohistochemical method, and the influence on chemosensitivity was analyzed through brain glioma U251 and U87 cell culture, in vitro drug (temozolomide) intervention and Transwell invasion in vitro. Results There was a significant difference in the expression of MGMT and Topo II genes in glioma specimens and normal brain tissues (P<0.05). The positive expression rates of MGMT gene in the two kinds of specimens were 63.75% and 3.33% respectively while the positive expression rates of Topo II gene were 55.00% and 0.00%, respectively (P<0.05). Both of MGMT and Topo II genes were displayed in the nucleus as positive staining. There was no significant difference in transmembrane cell count of U251 and U87 cells between the experimental group given drug intervention in vitro and negative control group without drug intervention before the intervention (P>0.05). However, the transmembrane cell count of U251 and U87 cells in the experimental group after intervention was higher than that in the negative control group (P<0.05). After intervetion, U251 and U87 cells were with stronger invasiveness in the experimental group. Conclusion The positive expression rates of MGMT and Topo II genes are high in human brain glioma specimens. They may be involved in promoting glioma cell invasion, affecting tumor chemosensitivity.

目的 探究叶酸对子宫内膜癌作用的靶基因。方法 通过转录组测序筛选叶酸作用下子宫内膜癌细胞中的差异基因,生存分析寻找对子宫内膜癌具有生存意义的差异基因,qPCR及western blot检测其在叶酸作用下的表达。结果 转录组测序发现36个差异基因,生存分析发现FMN1,TRIB3,INHBE及NRBP2的表达对子宫内膜癌具有生存意义,qPCR及western blot验证叶酸作用下NRBP2在子宫内膜癌细胞中的表达下调。结论 叶酸下调子宫内膜癌中NRBP2基因的表达,NRBP2可能是叶酸对子宫内膜癌作用的靶标。

Objective To explore the target genes of folic acid on endometrial carcinoma. Methods The differential genes in endometrial cancer cells treated with folic acid were screened by transcriptome sequencing. Survival analysis was used to find the differential genes with survival significance. QPCR and western blot were used to detect their expression under the action of folic acid. Results 36 differential genes were found by transcriptional sequencing. Survival analysis showed that the expression of FMN1,TRIB3,INHBE and NRBP2 had survival significance in endometrial carcinoma. QPCR and western blot confirmed that the expression of NRBP2 in endometrial cancer cells was down-regulated by folic acid. Conclusion Folic acid down-regulates the expression of NRBP2 gene in endometrial carcinoma, and NRBP2 may be the target of the effect of folic acid on endometrial carcinoma.