COOH-terminal deletion of HBx gene is a frequent event in HBV-associated hepatocellular carcinoma

AIM:To investigate the hepatitis B virus (HBV) x gene (HBx) state in the tissues of HBV-related hepatocellular carcinoma (HCC) in Chinese patients and whether there were particular HBx mutations. METHODS: HBx gene was amplified and direct sequencing was used in genomic DNA samples from 20 HCC and corresponding non-cancerous liver tissues from HBsAg-positive patients. HBV DNA integration and HBx deleted mutation were validated in 45 HCC patients at different stages by Southern blot analysis and polymerase chain reaction methods. RESULTS: The frequencies of HBx point mutations were significantly lower in HCC than their corresponding non- cancerous liver tissues (11/19 vs 18/19, P = 0.019). In contrast, deletions in HBx gene were significantly higher in HCC than their non-cancerous liver tissues (16/19 vs 4/19, P < 0.001). The deletion of HBx COOH-terminal was detected in 14 HCC tissues. A specific integration of HBx at 17p13 locus was also found in 8 of 16 HCC, and all of them also exhibited full-length HBx deletions. Integrated or integrated coexistence with replicated pattern was obtained in 45.5% (20/45) - 56.8% (25/45) tumors and 40.9% (18/45) - 52.3% (23/45) non-tumor tissues. CONCLUSION: HBx deletion, especially the COOH- terminal deletion of HBx is a frequent event in HBV-associated HCC tissues in China. HBV integration had also taken place in partial HCC tissues. This supporting the hypothesis that deletion and probably integrated forms of the HBx gene may be implicated in liver carcinogenesis.

HBV X Gene Transfection Upregulates IL-1β and IL-6 Gene Expression and Induces Rat Glomerular Mesangial Cell Proliferation

The X gene of HBV encodes a 17-kD protein, termed HBx, which has been shown to function as a transcriptional trans-activator of a variety of viral and cellular promoter/enhancer elements. The aim of this study was to investigate the effect of HBx on gene expression of interleukin （IL）-1β and IL-6, and proliferation of rat mesangial cells in vitro. The X gene of HBV was amplified by PCR assay, and inserted into the eukaryotic expression vector pCI-neo. The structure of recombinant pCI-neo-X plasmid was proved by restrict endonuclease digestion and sequencing analysis. pCI-neo-X was transfected into cultured rat mesangial cell line in vitro via liposome. HBx expression in transfected mesangial cells was detected by Western blot. The IL-1β and IL-6 mRNA expression in those cells was assayed by semiquantitative RT-PCR. Mesangial cell proliferation was tested by MTT. The results showed that HBx was obviously expressed in cultured mesangial cell line at 36th and 48th h after transfection. The expression of IL-1β and IL-6 mRNA was simultaneously increased. The cell proliferation was also obvious at the same time. It was concluded that HBx gene transfection could induce IL-1β and IL-6 gene expression and mesangial cell proliferation. HBx may play a critical role in mesangial cell proliferation through upregulation of the IL-1β and IL-6 gene expression.

Possible mechanism for hepatitis B virus X gene to induce apoptosis of hepatocytes

AIM： To investigate the possible mechanism for HBV X gene to induce apoptosis of hepatocyte HL-7702 cells.METHODS： HBV X gene eukaryon expression vector pcDNA3-X was established and transfected into HL-7702 cells by lipid-mediated transfection, including transient and stable transfection. Positive clones were screened by incubating in the selective medium with 600 μg/mL G418 and named HL-7702/HBV-encoded X protein （HBx） cells. The expressions of Fas/FasL, Bax/Bcl-2, and c-myc mRNA were measured by semi-quantitative RT-PCR in HL-7702/HBx and control group, respectively.RESULTS： RT-PCR analysis confirmed that HBV X gene was transfected into HL-7702 cells successfully. By semiquantitative RT-PCR analysis, Bax and c-myc mRNA levels in HL-7702/HBx cells of transient transfection were significantly higher than those in control, FasL and c-myc mRNA levels in HL-7702/HBx cells of stable transfection were significantly higher than those in control, whereas the Bcl-2 mRNA levels in HL-7702/HBx cells of transient and stable transfection were significantly lower than thosein control.CONCLUSION： HBV X gene may promote the apoptosis of hepatocytes by regulating the expressions of Fas/FasL, Bax/Bcl-2, and c-myc gene in a dose-dependent manner.

THE EXPRESSIONS OF HBV X GENE AND ets-2, IGF-Ⅰ, c-myc AND N-ras ONCOGENES IN HUMAN HEPATOCELLULAR CARCINOMA AND TUMOR-ADJACENT TISSUES

The expressions of HBV X gene and ets-2, IGF-I, c-myc and N-ras were studied in 7 pairs of human primary hepatocellular carcinoma (PHC) and tumor-adjacent tissues, using RNA hybridization and im-munoblot methods. The results showed that specific 17 and 28 kD HBV X gene products (HBxAg) were existed in a portion of PHC and tumor-adjacent tissues. The 17 kD HBxAg was detected in the sera of 3 patients who also had 17 kD HBxAg in their liver tissues. Multiple expressions of oncogenes such as ets-2, c-myc and N-ras were observed in PHC and tumor-adjacent tissues that had HBxAg expressed, indicating HBxAg might function as a transactivator in the course of intracellular proto-oncogene activation. It is also observed that in some tumor-adjacnet tissues the expressions of ets-2, c-myc and N-ras were higher than those in corresponding PHC. The relationship of HBxAg to the expression of est-2, IGF-Ⅱ, c-myc and their possible roles in the carcinogenesis of PHC are discussed.

Construction of B7x Gene Overexpression Lentiviral-based Vector

To construct overexpression lentiviral-based vector carrying rat B7x gene,B7x gene precursor sequences amplified by polymerse chain reaction（PCR） were ligated with pLVTHM to generate pLVTHM-B7x gene expression lentiviral-based vector.The positive clones were selected to be submitted to DNA sequencing.HEK293T cells were co-transfected with pLVTHM-B7x and two packaging plasmids psPAX2 and pMD2G to produce lentivirus which express B7x gene.The mRNA expression levels of B7x gene and virus titer were detected by real-time PCR in HEK293T cells.The B7x protein levels were detected by Western blot analysis in HEK293T cells.The identification of restriction enzyme digestion and DNA sequencing confirmed that pLVTHM-B7x lentiviral-based vector was successfully constructed.Green fluorescence was observed in HEK293T packaging cells by means of an inverted fluorescence microscope and the virus titer measured was 2×108 TU/mL.It will establish the foundation for studing deeply the biological function of B7x to construct successfully B7x expression lentiviral-based vector.

Study of Hepatitis B Virus Genotypes and Mutation in 1762 &1764 Nucleotides of X Gene in Chronic HBV Patients from Golestan Province—Iran

Introduction: More than 350 million people are chronic carriers of HBV and many of them develop progressive diseases, including cirrhosis and hepatocellular carcinoma. Many of those infected develop persistent disease and a proportion goes on to develop liver failure and cancer. Researchers showed that double mutations of the x gene at position 1762 and 1764, have been found in chronic hepatitis B. These mutations were proposed to be associated with fulminant hepatitis B increasing risk of hepatocellular carcinoma. This project aimed to investigate mutation in the x gene region of HBV infected patients in Golestan province, Iran. Method: 100 patients were entered in this study. Hepatitis B viral DNA was extracted from plasma and PCR was performed using specific primers. Direct sequencing and alignment of x gene were applied using reference sequence from Gene Bank database (Okamoto, 1988;Accession number AB033559). Results: Among the chronic HBV patients 51% were male. The results showed that 49% of patients had A1762T, G1764A mutations changing AGG to stop codon TGA. 27% and 24% of cases were showed mutation only in A1762T and G1764A positions respectively. Conclusion: This study was shown presence of X gene mutation in HBV infected people in Golestan province, Iran. The rate of mutation in two positions 1762 and 1764 of HBV genotype D X gene was higher than the average rate of the world (34%).

Detection of hyper-conserved regions in hepatitis B virus X gene potentially useful for gene therapy

AIM To detect hyper-conserved regions in the hepatitis B virus(HBV) X gene(HBX) 5' region that could be candidates for gene therapy.METHODS The study included 27 chronic hepatitis B treatmentnaive patients in various clinical stages(from chronic infection to cirrhosis and hepatocellular carcinoma, both HBeA g-negative and HBeA g-positive), and infected with HBV genotypes A-F and H. In a serum sample from each patient with viremia > 3.5 log IU/m L, the HBX 5' end region [nucleotide(nt) 1255-1611] was PCRamplified and submitted to next-generation sequencing(NGS). We assessed genotype variants by phylogenetic analysis, and evaluated conservation of this region by calculating the information content of each nucleotide position in a multiple alignment of all unique sequences(haplotypes) obtained by NGS. Conservation at the HBx protein amino acid(aa) level was also analyzed.RESULTS NGS yielded 1333069 sequences from the 27 samples, with a median of 4578 sequences/sample(2487-9279, IQR 2817). In 14/27 patients(51.8%), phylogenetic analysis of viral nucleotide haplotypes showed a complex mixture of genotypic variants. Analysis of the information content in the haplotype multiple alignments detected 2 hyper-conserved nucleotide regions, one in the HBX upstream non-coding region(nt 1255-1286) and the other in the 5' end coding region(nt 1519-1603). This last region coded for a conserved amino acid region(aa 63-76) that partially overlaps a Kunitz-like domain.CONCLUSION Two hyper-conserved regions detected in the HBX 5' end may be of value for targeted gene therapy, regardless of the patients' clinical stage or HBV genotype.

Characterization of hepatitis B virus X gene quasispecies complexity in mono-infection and hepatitis delta virus superinfection

Hepatitis delta virus(HDV) seems to strongly suppress hepatitis B virus(HBV)replication, although little is known about the mechanism of this interaction. Both these viruses show a dynamic distribution of mutants, resulting in viral quasispecies. Next-generation sequencing is a viable approach for analyzing the composition of these mutant spectra. As the regulatory hepatitis B X protein(HBx) is essential for HBV replication, determination of HBV X gene(HBX)quasispecies complexity in HBV/HDV infection compared to HBV monoinfection may provide information on the interactions between these two viruses.AIM To compare HBV quasispecies complexity in the HBX 5' region between chronic hepatitis delta(CHD) and chronic HBV mono-infected patients.METHODS Twenty-four untreated patients were included: 7/24(29.2%) with HBeAgnegative chronic HBV infection(CI, previously termed inactive carriers), 8/24(33.3%) with HBeAg-negative chronic hepatitis B(CHB) and 9/24(37.5%) with CHD. A serum sample from each patient was first tested for HBV DNA levels.The HBX 5' region [nucleotides(nt) 1255-1611] was then PCR-amplified for subsequent next-generation sequencing(MiSeq, Illumina, United States). HBV quasispecies complexity in the region analyzed was evaluated using incidencebased indices(number of haplotypes and number of mutations), abundancebased indices(Hill numbers of order 1 and 2), and functional indices(mutation frequency and nucleotide diversity). We also evaluated the pattern of nucleotide changes to investigate which of them could be the cause of the quasispecies complexity.RESULTS CHB patients showed higher median HBV-DNA levels [5.4 logIU/mL,interquartile range(IQR) 3.5-7.9] than CHD(3.4 logIU/mL, IQR 3-7.6)(P = n.s.)or CI(3.2 logIU/mL, IQR 2.3-3.5)(P < 0.01) patients. The incidence and abundance indices indicated that HBV quasispecies complexity was significantly greater in CI than CHB. A similar trend was observed in CHD patients, although only Hill numbers of order 2 showed statistically significant differences(CHB2.81, IQR 1.11-4.57 vs CHD 8.87, 6.56-11.18, P = 0.038). There were no significant differences in the functional indices, but CI and CHD patients also showed a trend towards greater complexity than CHB. No differences were found for any HBV quasispecies complexity indices between CHD and CI patients. G-to-A and C-to-T nucleotide changes, characteristic of APOBEC3 G, were higher in CHD and CI than in CHB in genotype A haplotypes, but not in genotype D. The proportion of nt G-to-A vs A-to-G changes and C-to-T vs T-to-C changes in genotype A and D haplotypes in CHD patients showed no significant differences. In CHB and CI the results of these comparisons were dependent on HBV genotype.CONCLUSION The lower-replication CHD and CI groups show a trend to higher quasispecies complexity than the higher-replication CHB group. The mechanisms associated with this greater complexity require elucidation.

Transcriptome profiling and RXR gene family identification reveals the molecular mechanism of rapid aging after spawning of cuttlefish Sepiella japonica

Sepiella japonica is a worldwide marine cuttlefish species of high economic value.S.japonica routinely modifying behaviors in reproductive life,such as rapid aging until death after spawning,has been recognized in artificial breeding.However,reproductive behavior at the level of genes is rarely reported,thus,the research on the genetic basis of behavior,reproduction,and artificial breeding was limited.We applied RNA-seq in different stages of reproduction to investigate the reason of rapid aging after spawning,pre-maturity,pre-spawning after maturity,and post-spawning.The retinoid X receptor(RXR)gene family in S.japonica was identified,and 1343–1452 differentially expressed genes(DEGs)in all 3 stages of reproductive life were identified from pairwise m RNA comparisons.Furthermore,through the GO term and KEGG analysis,S.japonica could handle neuronal development and network formation before maturity and have a functional degradation of neural communication,signal transduction,vision,and gene expression after spawning.Eight Sj RXRαs have been identified and they played different roles in growth development or reproduction.Therefore,the regulation of several channels and receptors is the intrinsic molecular mechanism of rapid aging after spawning in S.japonica.This study revealed the survival strategy and provided fundamental data on the level of genes for understanding the reproductive behavior and the reproduction of S.japonica.

失活X染色体基因逃逸与系统性红斑狼疮的性别二态性

X染色体失活可平衡女性中两条X染色体的基因剂量。越来越多的证据表明,失活X染色体上存在许多能够逃逸失活的基因。逃逸的机制涉及到DNA、RNA、组蛋白的表观修饰以及众多的调控蛋白和染色质的空间结构。失活X染色体基因逃逸的研究为人类疾病(特别是自身免疫性疾病)性别二态性的研究开辟了新的途径。目前已证实包括TLR7、CD40L、IRAK-1、CXCR3、CXorf21等失活X染色体基因逃逸是系统性红斑狼疮(systemic lupus erythematosus,SLE)女性好发的重要原因。本文主要综述了失活X染色体上基因逃逸以及与SLE性别二态性形成的分子机制。阐明SLE性别二态性形成的分子机制,不仅对疾病的诊断、治疗具有重要意义,而且对深入揭示人类免疫系统的发育及调控机理也有重要的理论意义。

Gene X-pert MTB/RIF检测对肺结核病的诊断价值研究

目的 研究分析肺结核病应用Gene X-pert MTB/RIF检测的诊断价值。方法 选取2022年3—10月在本院就诊的疑似肺结核病患者164例为研究对象,所有患者均给予结核分枝杆菌培养检查、痰涂片抗酸染色法诊断、Gene X-pert MTB/RIF检测,以结核分枝杆菌培养检查结果为金标准,比较痰涂片抗酸染色法诊断与Gene X-pert MTB/RIF检测的阳性率以及诊断准确性、灵敏度、特异度。结果 痰涂片抗酸染色法诊断阳性率为21.34%(35/164),Gene X-pert MTB/RIF检测阳性率为35.98%(59/164),Gene X-pert MTB/RIF检测明显高于痰涂片抗酸染色法诊断(P<0.05)。痰涂片抗酸染色法诊断的准确性为85.37%,灵敏度为59.65%,Gene X-pert MTB/RIF检测的准确性为95.12%,灵敏度为94.74%,Gene X-pert MTB/RIF检测明显高于痰涂片抗酸染色法诊断(P<0.05)。结论 在肺结核病诊断中,Gene X-pert MTB/RIF检测的阳性率更高,同时诊断准确率与灵敏度也更高,为肺结核病的诊断与治疗提供了可靠的指导依据。

PXR基因单核苷酸多态性与2型糖尿病患病风险的关系

目的探讨孕烷X受体(PXR)基因单核苷酸多态性(SNP)与2型糖尿病(T2DM)患病风险的关系。方法选择T2DM患者285例(观察组)、同期体检健康的志愿者230例(对照组),采集所有研究对象空腹外周静脉血,提取基因组DNA,然后对PXR基因rs1523127、rs3814055、rs6785049位点进行测序和基因分型;采用ELISA法检测血清PXR、葡萄糖转运体2(GLUT2)、葡萄糖激酶(GCK)。比较两组PXR基因rs1523127、rs3814055、rs6785049位点基因型及等位基因频率,以及血清PXR、GLUT2、GCK水平。分析PXR基因SNP与T2DM患病风险的关系。结果经Hardy-Weinberg遗传平衡检验,两组PXR基因不同位点基因型、等位基因频率均符合遗传平衡定律。两组PXR基因rs1523127、rs6785049位点基因型及等位基因频率比较差异均无统计学意义(P均>0.05)。观察组PXR基因rs3814055位点CT/TT基因型及T等位基因频率均高于对照组(P均<0.05),携带CT、TT基因型者罹患T2DM的优势比(OR)分别为携带CC基因型者的1.591、2.398倍,携带T等位基因者罹患T2DM的OR为携带C等位基因者的1.638倍。观察组血清PXR水平高于对照组,血清GLUT2、GCK水平低于对照组(P均<0.05)。T2DM患者PXR基因rs3814055位点CT/TT基因型者血清PXR水平高于CC基因型者,血清GLUT2、GCK水平低于CC基因型者(P均<0.05)。结论PXR基因rs3814055位点C等位基因突变为T等位基因能够增加其转录活性,抑制血清GLUT2、GCK水平,使其糖耐量受损,进而增加T2DM的患病风险。

转导蛋白β样1X基因与迟发性感音神经性耳聋

目前的研究表明,人源转导蛋白β样1X(transducin beta like 1 X-linked,TBL1X)基因与多种疾病的发生密切相关,还参与了迟发性感音神经性耳聋的发生,我们回顾国内外的相关文献,就该基因与迟发性感音神经性耳聋的相关研究做一综述。

复方斑蝥胶囊联合旋转容积调强技术对头颈部放疗生存期及血清XRCC1、XRCC3mRNA水平的影响

目的探究复方斑蝥胶囊联合旋转容积调强技术对头颈部放疗患者生存期及血清X线修复交错互补基因1(X-ray Repair Cross Complementing 1,XRCC1)、X线修复交错互补基因3(X-ray Repair Cross Complementing 3,XRCC3)信使核糖核酸(Messenger RNA,mRNA)水平的影响。方法选取2019年6月—2021年7月医院收治的头颈部肿瘤患者97例作为研究对象,根据治疗方法不同进行分组,对照组(48例)采用旋转容积调强技术结合放疗治疗,联合组(49例)在对照组基础上加用复方斑蝥胶囊治疗。观察比较临床疗效、中医证候积分、血清XRCC1及XRCC3mRNA水平、生存期、不良反应。结果联合组客观缓解率高于对照组(P<0.05),但疾病控制率与对照组比较差异无统计学意义(P>0.05)。联合组自汗、恶心呕吐、神疲乏力、食欲差、失眠、头晕眼花等得分低于对照组(P<0.05)。联合组血清XRCC1、XRCC3水平及外周血XRCC1、XRCC3 mRNA表达均低于对照组(P<0.05)。联合组无进展生存期和总生存期均高于对照组(P<0.05)。联合组不良反应发生率(34.69%,17/49)低于对照组不良反应发生率(72.92%,35/48)(P<0.05)。结论复方斑蝥胶囊联合旋转容积调强技术能改善头颈部放疗患者肿瘤客观缓解率,缓解临床症状,降低血清XRCC1、XRCC3mRNA水平,减少不良反应。

伴有癫痫的脆性X综合征家系1例

脆性X综合征(fragile X syndrome,FXS)是FMR1基因CGG异常重复扩增导致的疾病。本文报告1对经基因检测诊断为FXS的兄弟,2例患者分别为15岁和14岁,均存在语言障碍、智力障碍、注意力缺陷障碍、孤独症谱系障碍和FXS特征性面容等临床表现,其中先证者伴有罕见的晚发性癫痫发作,经左乙拉西坦治疗效果良好,而其弟弟经反复随访未见脑电图异常。该对病例提示FXS临床表型具有多样性和异质性。

烟草粉螟卵黄原蛋白基因鉴定及其对X射线胁迫的响应

【背景和目的】卵黄原蛋白(vitellogenin,Vg)是昆虫卵黄发生的关键物质,为探索X射线辐照胁迫对烟草粉螟Vg基因转录表达和雌成虫繁殖力的影响。【方法】采用RT-PCR技术克隆烟草粉螟Vg基因的全长并进行生物信息学分析,利用qPCR技术检测在烟草粉螟雌成虫不同组织(足、头、中肠、脂肪体、卵巢和表皮)的表达量和雌成虫在不同X射线辐照剂量(50,100和150 Gy)胁迫下该基因的相对表达量以及对其蛹羽化率、成虫寿命、产卵量、卵孵化率的影响。【结果】(1)克隆获得烟草粉螟卵黄原蛋白基因,命名为EeVg(GenBank登录号:OM804251),开放阅读框(ORF)长度为5337bp,编码1778个氨基酸。(2)EeVg基因在雌成虫的脂肪体中相对表达量最高,为头部表达量的17.96倍。(3)100和150 Gy的X射线辐照雌蛹极显著地降低EeVg在烟草粉螟羽化后的表达量,分别为对照的0.34倍和0.09倍。(4)剂量为50、100和150 Gy的X射线辐照降低烟草粉螟雌蛹羽化率和成虫寿命(87.78%~75.56%和6.40~4.17d),显著降低雌蛾的产卵量和卵孵化率(55.27~20.37粒和76.67%~72.22%)。【结论】高剂量的X射线辐照胁迫可使EeVg基因表达量显著下调,显著降低烟草粉螟繁殖力,为深入研究X射线影响烟草粉螟繁殖力的分子机制奠定了基础。

乙型肝炎病毒X基因通过抑制Caspase-1表达促进肝癌细胞增殖

目的 探究乙型肝炎病毒X基因(HBx)对HepG2细胞中Caspase-1介导的肝细胞增殖的影响。方法 用脂质体转染法将HBx真核表达载体pCDNA3.1-HBx瞬时转入HepG2细胞,以阴性载体转染的HepG2细胞(NC)为对照。转染后72 h收集细胞,通过RT-PCR检测HBx的表达,通过qPCR和Western blot法检测Caspase-1的mRNA和蛋白质表达,并通过CCK-8检测HepG2-HBx和HepG2-NC的增殖情况。结果 RT-PCR证实HBx成功转染HepG2细胞株,HepG2-HBx细胞中Caspase-1的mRNA和蛋白表达水平显著低于HepG2-NC组。HepG2-HBx细胞的增殖能力显著高于HepG2-NC组。结论 HBx通过抑制Caspase-1的表达诱导HepG2细胞增殖。

一个新PHEX基因变异导致X连锁遗传性低血磷性佝偻病的家系遗传学分析

目的以一个临床表型为低血磷性佝偻病(hypophosphatemic rickets,HR)的家系为研究对象,通过全外显子组测序寻找该家系的致病变异基因,并分析变异的致病性。方法收集HR家系临床资料,进行生化检测。提取先证者DNA,进行临床全外显子组测序,并针对可疑致病变异对家系所有成员进行PCR扩增,Sanger测序验证。预测变异的致病性及对蛋白质空间结构的影响。结果该家系共3代,先证者是一位26岁女性,先证者母亲、先证者及其儿子和女儿为患者,临床表现为O型腿、鸡胸、低磷血症,骨骼X线检查、生化检测、成纤维细胞生长因子23(fibroblast growth factor 23,FGF23)检测结果提示低血磷性佝偻病。临床全外显子组测序发现PHEX(NM_000444)c.2193dupT的插入移码杂合变异,该变异可导致编码第732位的天冬酰胺变异为终止密码子(p.N732*),从而出现了蛋白质截短。先证者的母亲、儿子及女儿PHEX基因均存在该变异。根据美国医学遗传学与基因组学学会(American College of Medical Genetics and Genomics,ACMG)对变异的分类标准,分级为可能致病性变异。经查阅文献及查找人类基因突变数据库,该变异均未被报道或收录。结论本研究发现了一个PHEX新致病变异,为该家系的临床诊断和治疗及遗传咨询提供了实验依据。

血清SIRT1、Fibulin-5、Bcl-2/Bax与颈动脉粥样硬化斑块破裂所致脑梗死的关系及联合检测价值

目的 探讨血清沉默信息调节蛋白1 (SIRT1)、衰老关键蛋白抗原-5 (Fibulin-5)、B淋巴细胞瘤基因-2(Bcl-2)/B淋巴细胞瘤基因-2相关X蛋白(Bax)与颈动脉粥样硬化(CAS)斑块破裂所致脑梗死(ACI)的关系及联合检测价值。方法 选取新疆维吾尔自治区人民医院2021年1月至2023年2月CAS斑块破裂所致ACI患者98例作为研究组,另选取同期CAS斑块未破裂患者98例作为对照组,比较两组血清SIRT1、Fibulin-5、Bcl-2、Bax水平,分析各血清指标对CAS斑块破裂所致ACI风险的影响及与病情的关系,并评价各血清学指标单独及联合预测CAS斑块破裂所致ACI的价值。结果 研究组血清SIRT1、Bcl-2水平低于对照组,Fibulin-5、Bax水平高于对照组(P<0.05);大面积梗死(MCI)患者血清SIRT1、Bcl-2水平<小面积梗死患者<腔隙性梗死(LI)患者,Fibulin-5、Bax水平>小面积梗死患者> LI患者(P<0.05);重度神经功能缺损患者血清SIRT1、Bcl-2水平<中度神经功能缺损患者<轻度神经功能缺损患者,Fibulin-5、Bax水平>中度神经功能缺损患者>轻度神经功能缺损患者(P<0.05);血清SIRT1、Bcl-2低水平患者CAS斑块破裂所致ACI风险是高水平患者的2.311倍、2.921倍,Fibulin-5、Bax高水平患者CAS斑块破裂所致ACI风险是低水平患者的3.470倍、3.184倍(P<0.05);血清SIRT1、Bcl-2与梗死面积、神经功能缺损程度呈负相关,Fibulin-5、Bax与梗死面积、神经功能缺损程度呈正相关(P<0.05);血清SIRT1、Fibulin-5、Bcl-2、Bax预测CAS斑块破裂所致ACI的AUC分别为0.716 (95%CI:0.648~0.778)、0.796 (95%CI:0.733~0.850)、0.728 (95%CI:0.660~0.789)、0.763 (95%CI:0.698~0.821),联合预测CAS斑块破裂所致ACI的AUC为0.909 (95%CI:0.860~0.945),优于各血清指标单独预测。结论 血清SIRT1、Fibulin-5、Bcl-2/Bax与CAS斑块破裂所致ACI及其病情程度密切相关,联合预测价值可靠,对临床开展防治工作具有指导意义。

IL-2RG基因c.675 C>A突变引起X-连锁重症联合免疫缺陷病患儿的基因检测及实验室与临床结果分析

目的探讨白细胞介素-2受体共同r链(interleukin 2 receptor gamma,IL-2RG)基因新发变异导致患儿重症联合免疫缺陷病(severe combined immunodeficiency,SCID)的分子遗传学特点和临床特征。方法分析西北妇女儿童医院儿童血液内科收治的一例重症联合免疫缺陷病患儿的临床资料、实验室结果及基因检测数据。结果一例两个月男婴,出生后反复感染入院治疗,患儿血细胞检测结果提示白细胞总数正常,但淋巴细胞明显减少;淋巴细胞亚群结果显示总T(CD3+),辅助T(CD3+CD4+),杀伤T(CD3+CD8+)和NK(CD3-CD16+CD56+)淋巴细胞占比明显减低,而B(CD3-CD19+)淋巴细胞占比明显升高;IgG明显减低,IgM和IgA小于检测下限;该患儿在感染状态下细胞因子水平未明显升高。患儿母亲家系中近3代有9例男性在出生后1岁内因反复感染致死,核心家系全外显子组测序结果发现,患儿X染色体IL-2RG基因(chrX:70329160)存在半合新发错义突变[c.675 C>A,p.S225R(p.Ser225Arg)],其母亲为携带者。依据以上证据患儿被确诊为X连锁重症联合免疫缺陷病(X-SCID)。随后每月静脉注射免疫球蛋白并服用常规抗生素和抗病毒药物预防感染,为患者造血干细胞移植做准备。因患儿出生后已接种卡介苗,患儿6月龄出现了播散性卡介苗病。经治疗后行造血干细胞移植。结论X-SCID患儿机体免疫功能缺陷严重,危及生命,接种活疫苗可能导致严重感染;该研究发现IL-2RG基因c.675 C>A突变是先证者X-SCID遗传学病因的新发致病变异,扩充了X-SCID致病基因IL-2RG的基因变异谱。