血清CysC、α-Syn联合MRI对帕金森病病情评估及诊断的意义

目的 探讨血清胱抑素C(CysC)、α-突触核蛋白(α-Syn)联合磁共振成像(MRI)对帕金森病(PD)病情评估及诊断的意义。方法 将2019年4月至2021年4月于河北省沧州中西医结合医院(下称该院)治疗的PD患者87例纳入研究作为PD组。根据Hoehn-Yahr(H-Y)分期,将PD患者分为Ⅰ~Ⅱ期13例,Ⅲ~Ⅳ期45例,Ⅴ期29例。另外,选取同期于该院进行体检的健康志愿者87例作为对照组。比较各组血清CysC、α-Syn水平,分析血清CysC、α-Syn联合MRI对PD的诊断效能。结果 PD组血清CysC、α-Syn水平均高于对照组,差异有统计学意义(P<0.05);血清CysC、α-Syn水平随着H-Y分期递增升高(P<0.05)。PD组MRI各相位值与对照组比较,黑质致密带、苍白球、壳核、尾状核相位值均降低(P<0.05);不同H-Y分期的PD患者黑质致密带、苍白球、壳核相位值比较,差异均有统计学意义(P<0.05)。CysC、α-Syn单独诊断PD的AUC分别为0.786、0.767,灵敏度分别为58.62%、64.37%,特异度分别为88.51%、87.36%;与MRI、血清CysC、α-Syn单独诊断相比,三项联合诊断PD的灵敏度和准确度较高同,差异有统计学意义(P<0.05)。结论 血清CysC、α-Syn联合MRI对PD诊断效能较高,能为评估PD病情严重程度和早期诊断提供依据。

血清生长相关蛋白-43、α-突触核蛋白对小儿癫痫诊断价值研究

目的探讨癫痫患儿血清生长相关蛋白-43(GAP-43)、α-突触核蛋白(α-Syn)水平,分析两者对小儿癫痫的诊断价值。方法纳入2019年4月—2022年4月临汾市人民医院儿科诊治小儿癫痫患者80例为癫痫组,晕厥患儿50例为晕厥组,腹股沟斜疝患儿50例为对照组。利用酶联免疫吸附试验检测血清GAP-43、α-Syn水平;比较3组患儿间及不同临床特征癫痫患儿血清GAP-43、α-Syn水平差异;Pearson相关分析血清GAP-43、α-Syn与癫痫发作严重程度量表评分(NHS3)的相关性;受试者工作特征曲线分析血清GAP-43、α-Syn对小儿癫痫的诊断价值。结果血清GAP-43水平比较,癫痫组<晕厥组<对照组(F/P=821.793/<0.001),血清α-Syn水平比较,癫痫组>晕厥组>对照组(F/P=419.176/<0.001);癫痫患儿血清GAP-43在癫痫局灶性发作、无认知功能损害者中升高(t/P=8.745/<0.001,10.070/<0.001),α-Syn水平在癫痫局灶性发作、无认知功能损害者中降低(t/P=4.236/<0.001,14.881/<0.001),二者在患儿性别、年龄、脑电图异常、头颅MR异常者中比较,差异无统计学意义(P均>0.05);血清GAP-43水平与NHS3评分呈显著负相关(r/P=-0.645/<0.001),血清α-Syn水平与NHS3评分呈显著正相关(r/P=0.702/<0.001);血清GAP-43、α-Syn及两项联合预测小儿癫痫的AUC分别为0.740、0.738、0.835,二项联合的AUC高于单项预测(Z=4.482、4.391,P均<0.001)。结论癫痫患儿血清GAP-43降低,α-Syn水平升高,两者与癫痫类型、认知功能损害有关,两项联合对小儿癫痫具有较高的诊断价值。

通用流感mRNA候选疫苗的构建及免疫效果评价

目的构建通用流感mRNA疫苗并全面评价其免疫保护效果。方法优化流感病毒株A/California/04/2009的血凝素(hemagglutinin,HA)、核蛋白(nucleoprotein,NP)和基质蛋白2胞外区(matrix protein 2 ectodomain,M2e)抗原序列,并将HA、NP和3个串联的M2e(3M2e)分别克隆至pcDNA3.1载体,通过线性化、体外转录、酶学法加帽、酶促加尾合成mRNA,命名为mRNA-HA、mRNA-NP和mRNA-3M2e。3种mRNA分别转染293T细胞后,通过免疫荧光实验鉴定蛋白的表达。利用脂质纳米颗粒分别包裹mRNA-HA、mRNA-NP和mRNA-3M2e并测量其粒径和电位。再将3种mRNA等体积混合制备成Comb-mRNA疫苗。将28只6周雌性Balb/c小鼠(体质量为18~22 g)按简单随机分组法分为2组:LNP组(n=14)和Comb-mRNA组(n=14)。通过血凝抑制(hemagglutination inhibition,HI)、微量中和(microneutralization,MN)实验评价流感mRNA疫苗诱导小鼠产生的血清抗体滴度;通过流式细胞术评价Comb-mRNA疫苗诱导的细胞免疫反应。采用5LD_(50)野生型H1N1流感病毒株感染小鼠评价Comb-mRNA疫苗的免疫保护效果。结果成功构建mRNA-HA、mRNA-NP和mRNA-3M2e,且3种mRNA均能在293T细胞表达。脂质纳米颗粒包裹mRNA平均粒径为(119.53±6.5)nm,平均电位为(-8.23±1.3)mV。与LNP组比较,Comb-mRNA组疫苗的HI几何平均滴度(geometric mean titer,GMT)达179.6,MN的GMT达201.6,同时诱导IFNγ+CD4+/CD8+T细胞比例升高。Comb-mRNA组在加强免疫后2周能够提供对5LD_(50)野生流感H1N1亚型病毒的保护。结论通用流感疫苗候选疫苗Comb-mRNA能够诱导小鼠免疫反应并保护小鼠免受病毒感染。

TP53INP1介导TGF-β1对膀胱癌细胞增殖、迁移的影响

目的 分析肿瘤蛋白53诱导的核蛋白1(TP53INP1)介导转化生长因子-β1(TGF-β1)对膀胱癌细胞增殖、迁移的影响。方法 选取2019年3月至2022年3月膀胱癌切除术的患者癌组织、癌旁组织标本40例,检测TP53INP1、TGF-β1表达量。膀胱癌细胞传代培养后分为TP53INP1-NC组、TP53INP1组、TGF-β1-NC组、TGF-β1组、TP53INP1+anti-TGF-β1-NC组、TP53INP1+anti-TGF-β1组。分析各组细胞增殖、凋亡、迁移情况。结果 与癌组织比较,癌旁组织TP53INP1、TGF-β1表达量较低(P<0.05)。与TP53INP1-NC组比较,TP53INP1组TP53INP1、B淋巴细胞瘤-2相关X蛋白(Bax)、天冬氨酸特异性半胱氨酸蛋白酶3(Caspase-3)、上皮性钙黏附素(E-cadherin)表达量、凋亡率水平较高,增殖率、细胞迁移数水平、B淋巴细胞瘤2基因(Bcl-2)、基质金属蛋白酶-2(MMP-2)、基质金属蛋白酶-9(MMP-9)表达量较低(P<0.05)。与TGF-β1-NC组比较,TGF-β1组TGF-β1、Bax、Caspase-3、E-cadherin表达量、凋亡率水平较高,增殖率、细胞迁移数水平、Bcl-2、MMP-2、MMP-9表达量较低(P<0.05)。双荧光素酶报告试验显示,与TGF-β1-NC组比较,TGF-β1组WT-TP53INP1荧光素酶活性降低(P<0.05),对MUT-TP53INP1荧光素酶活性影响较小(P>0.05)。与TP53INP1+anti-TGF-β1-NC组比较,TP53INP1+anti-TGF-β1组TGF-β1、Bax、Caspase-3、E-cadherin表达量、凋亡率水平较低(P<0.05),增殖率、细胞迁移数水平、Bcl-2、MMP-2、MMP-9表达量较高(P<0.05)。结论 TP53INP1可能通过靶向作用于TGF-β1而参与膀胱癌细胞增殖、凋亡、迁移,为膀胱癌诊治提供了新方向。

Nucleoprotein-based indirect enzyme-linked immunosorbent assay(indirect ELISA) for detecting antibodies specific to Ebola virus and Marbug virus

Full-length nucleoproteins from Ebola and Marburg viruses were expressed as His-tagged recombinant proteins in Escherichia coli and nucleoprotein-based enzyme-linked immunosorbent assays(ELISAs) were established for the detection of antibodies specific to Ebola and Marburg viruses. The ELISAs were evaluated by testing antisera collected from rabbit immunized with Ebola and Marburg virus nucleoproteins. Although little cross-reactivity of antibodies was observed in antiEbola virus nucleoprotein rabbit antisera, the highest reactions to immunoglobulin G(Ig G) were uniformly detected against the nucleoprotein antigens of homologous viruses. We further evaluated the ELISA's ability to detect antibodies to Ebola and Marburg viruses using human sera samples collected from individuals passing through the Guangdong port of entry. With a threshold set at the mean plus three standard deviations of average optical densities of sera tested, the ELISA systems using these two recombinant nucleoproteins have good sensitivity and specificity. These results demonstrate the usefulness of ELISA for diagnostics as well as ecological and serosurvey studies of Ebola and Marburg virus infection.

Preparation and Initial Application of a Monoclonal Antibody Specific for a Newly Discovered Conserved Linear Epitope of Rabies Virus Nucleoprotein

Objective To prepare monoclonal antibodies against a newly discovered and conserved linear epitope of Rabies virus nucleoprotein and to use them in a rabies diagnostic test. Methods Synthetic peptide containing the epitope was used as immunogen to prepare hybridoma cell lines by classical hybridoma technology. Anti-peptide monoclonal antibodies produced in ascites of inoculated Balb/c mice were labeled with fluorescein isothiocyanate （FITC） after purification and used in fluorescent antibody test （FAT）. Results Two positive hybridoma cell lines, RVNP-mAbl-CL and RVNP-mAb2-CL, were obtained. RVNP- mAbl-CL produced a higher concentration of monoclonal antibody RVNP-mAbl in Balb/c ascites. FITC-labeled RVNP-mAbl showed correct results on certain Rabies virus-positive canine brain tissue samples and cells of a small subclone of baby hamster kidney 21 cell line （BSR）. Conclusion FITC-labeled RVNP-mAbl has potential application for laboratory diagnosis of rabies

Expression of Nucleoprotein Gene of CTN Strain Rabies Virus from China in <i>E. coli </i>with Antigenicity

The nucleoprotein (NP) gene of rabies CTN strain isolated from China was recombined into pMal-c2x. The antigenicity of the recombined MBP-NP fusion proteins was examined by western blotting and by enzyme linked immunosorbent assay (ELISA). The results demonstrated that the recombined protein possesses predominant antigenicity.

Evaluation of Benzamide Derivatives as New Influenza A Nucleoprotein Inhibitors

Virus nucleoprotein (NP) is an emerging target for drug development for Influenza. We designed benzamide derivatives as new inhibitors of NP that demonstrate good potency in blocking influenza A. Screening revealed that compound 39 was the most potent molecule in the series, exhibiting IC50 values of 0.46 and 0.27 μM in blocking the replication of H3N2 (A/HK/8/68) and (A/WSN/33) influenza A viral strains. The observed inhibition of viral replication correlated well with cytopathic protection. Furthermore, based on computational analysis and fluorescence microscopy, it was determined that compound 39 inhibited nuclear accumulation by targeting influenza A viral nucleoproteins. Finally, the rodent pharmacokinetic profile of compound 32 displayed half-life of greater than 4 hours and bioavailability greater than 20%, suggesting this class of molecules had drug-like properties.

Phosphorylation sites within Ebola virus nucleoprotein

To understand the infection process, the viral multiplication and entry to the cell is widely studied. The Ebola virus nucleoprotein is the important problem for the pathological process. Focusing on the specific biological process, the post translational modification is needed. Here, the authors used the bioinformatics study to find the phosphorylation sites within the Ebola virus nucleoprotein and could identify many new sites.

Physiological Effects of Salmon Milt Nucleoprotein on Movement, Stress Tolerance and Lifespan of <i>C. elegans</i>

In recent years, various physiological functions of salmon milt extract, which consists of nucleic acid and nucleoprotein, have been reported. The objective of this study is to analyze the physiological function and its mechanism of salmon milt extract (NG) on nematodes (C. elegans). The wild type nematode N2 strain was bred on the plate containing of NG for four days, and its body length increased depending on NG concentration. When nematodes were bred with NG for a longer period, average lifespan was increased, and survival rate was increased by up to 20%. Generally, the movement of nematodes decreases with longer breeding period (i.e. aging). Analysis of movement (both gross thrashing movement and local pumping movement) showed that NG suppressed this decrease f movement with aging. Furthermore, the deease of survival rate by heat stress and oxidative stress was suppressed by NG administration. Nile Red staining analysis showed that fat accumulation varied depending on the concentration of NG. RT-PCR analysis revealed that the mRNA expression levels of the stress resistance genes sod-3 and sod-4 were increased. These results indicated that NG administration increased the expression of stress-tolerance-related genes, promoted stress tolerance, increased movement and prolonged lifespan in nematode.

SSINCC: Simple separation of interacting nucleoprotein complex components

Protein-DNA binding assays have been used in a va-riety of applications from fundamental studies re-garding the binding process itself to serve as probes for the detection, quantification and separation of target analytes. Here we describe a novel method of analyzing and identifying intermolecular DNA interactions that allows for the simple separation of interacting nucleoprotein complex components (SSINCC), focusing specifically on DNA-DNA interactions using P1 plasmid active partition system nucleoprotein complexes as a model to demonstrate DNA sequence specificity and tolerance of composite factor complexity. Traditional and recent assays of protein-DNA interaction are summarized and compared with SSINC. Although SSINC is examined here employing P1 partition nucleoprotein complex as an example of DNA-DNA intermolecular association, universal applications of this methodology to nucleo-protein complex studies can be envisioned.

Immunoglobulin G Specific Antibody Level against Ebola Viral Glycoprotein and Nucleoprotein in Ebola Virus Disease Survivors and Their Relatives

Ebola virus disease is a complex zoonosis that is highly virulent in humans. Despite its sorely pathogenic and lethal nature, survivors of this infection and even asymptomatic cases are able to develop both humoral and cellular immunity against several Ebola virus (EBOV) proteins. We aimed at determining immunoglobulin G (IgG) antibodies level against two Ebola viral antigens, the glycoprotein and the nucleoprotein in Ebola survivors and their relatives. Anti-EBOV glycoprotein (GP) and nucleoprotein (NP) IgG antibodies were quantified using ELISA. We enrolled 199 participants in two different sites as follow: 91 survivors at the Loreto clinic and 70 survivors with 38 relatives of Sierra Leone Association of Ebola Survivors Bombali Branch (SLAESB) tested for anti-EBOV NP and anti-EBOV GP IgG antibodies. Our findings revealed that the median anti-EBOV IgG level among survivors was 5.7128 U/ml [IQR: 2.793 - 7.783] for anti-EBOV GP IgG and 4.431 U/ml [IQR: 2.083 - 7.696] for anti-EBOV NP IgG. Survivors relatives had a median anti-EBOV GP IgG level of ?0.7128 U/ml [IQR: -0.903 to -0.04327] and -2.711 U/ml [IQR: -4.01 to -1.918] for anti-EBOV NP IgG. We observed that IgG levels in survivors were higher than in relatives with a significant difference of about 0.0001. The median value of anti-EBOV IgG level among seropositive relatives was 0.7043 U/ml [IQR: 0.5686 to 3.716] for anti-EBOV GP IgG and 4.05 U/ml [IQR: 0.2765 to 7.759] for anti-EBOV NP IgG respectively. Interestingly, we observed that 3.30% of Loreto clinic survivors did not developed anti-EBOV NP IgG antibodies;also about 10% survivors of the SLAESB were not reactive to anti-EBOV NP IgG and 1.43% of these survivors did not express antibodies against the Ebola viral glycoprotein. Our work is consistent with previous published studies showing heterogeneity in both survivors and asymptomatic cases of Ebola infection developing adaptive immunity against EBOV proteins.

Sequencing, Expression and Diagnostic Application of the Nucleoprotein Gene of Xinjiang Hemorrhagic Fever Virus

In order to analyze the nucleoprotein (NP) gene of Crimean-Congo hemorrhagic fever virus (CCHFV), viral RNA was amplified by RT-PCR by using the proof-reading DNA polymerase to produce the complete NP gene. The PCR product was sequenced, analyzed for phylogenesis and cloned into the expression vector pET32a and the recombinant plasmid expressed in E.coli BL-21 with high yield. The primarily purified fused protein was used to coat ELISA plates for the detect antibodies. It was found the similarities between NP gene of BA88166 and other XHFVs in nucleotide level and amino acid contents were very significant, and the NP gene of BA88166 encoded a nucleoprotein with 482 amino acid and a deduced molecular weight (MW) of 54?kDa. Western blot assay showed that the fusion protein expressed in bacteria possessed good antigenicity. The results with ELISA for the detection of the human and animal sera collected in endemic areas were found to be in good accordance to the clinical diagnosis. It concluded that the relations of NP genes of XHFV BA88166 and other XHFVs appeared to be evolutionally close. The methodologies established in this study were accurate, specific, rapid and reproducible for the clinical examinations and epidemiological survey.

CLINICAL SIGNIFICANCE OF DETECTING SERUM ANTIBODIES TO NUCLEOPROTEIN AND GLYCOPROTEIN G_2 OF HANTAAN VIRUS IN PATIENTS WITH HEMORRHAGIC FEVER WITH RENAL SYNDROME

Antibody blocking enzyme linked immunosorbent assays, respectively detecting antibodies to Hantaan virus nucleoprotein (NPAb) and glycoprotein GZ (G,Ab), were developed using monoclonal antibody L133, L13r3, LV48A and LVZB28B NPAb and GZAb in 291 serum samples from 65 patientswith kemorrkagic fever with renal syudrome (HFRS) were detfrmlned by these methods. The positive rates or NPAb were 90N on day 2-3 and 100 % on day 8-9 arter onset of disease, respectively.NPAb titers Increased during fever period and reached Peak levels during kypotensive and oliguric periods of HFRS. It was suggested that NPAb might be an important component Involved in the immunopathogenlc lin'alrmeut of HFRS and the detection of NPAb might be useful for the early diagnosis or HFRS. The I,osltlve rates and titers of GZAh were very low during the rirst three periods,namely rever, hypoteuslve and ollgurlc periods, and reached high levels during the convalescent period. GRAb titers were negatively related to the I,rotelnurla levels during the course of HFRS. It wasIndicated that GZAb might be the main component or neutralizing autlhodles to Hantaan virus Infection and the efrlclent production or GZAb was a good marker ror predicting the recovery and betterprognosis of HFRS.

动脉瘤蛛网膜下腔出血后迟发性脑缺血患者血清水通道蛋白4、神经元特异性核蛋白水平及意义

目的探讨动脉瘤蛛网膜下腔出血(aSAH)后迟发性脑缺血患者血清水通道蛋白4(AQP4)和神经元特异性核蛋白(NeuN)水平变化及临床意义。方法选取100例aSAH患者为研究对象,根据aSAH后迟发性脑缺血发生情况将患者分为无迟发性脑缺血组(n=68)和迟发性脑缺血组(n=32)。血清中AQP4和NeuN水平采用酶联免疫吸附法(ELISA)检测,采用Pearson法分析血清AQP4与NeuN的相关性。采用受试者工作特征(ROC)曲线分析血清AQP4和NeuN水平对aSAH后迟发性脑缺血的预测价值。结果与无迟发性脑缺血组相比,迟发性脑缺血组WFNS分级为Ⅲ~Ⅳ级患者占比、Hunt-Hess分级为Ⅲ~Ⅳ级占比以及血清AQP4水平较高,血清NeuN水平较低,差异有统计学意义(P<0.05)。Pearson法分析显示,血清AQP4和NeuN呈负相关(P<0.05)。Logistic回归分析显示,Hunt-Hess分级、AQP4是aSAH后迟发性脑缺血的危险因素,NeuN是aSAH后迟发性脑缺血的保护因素(P<0.05)。ROC曲线显示,AQP4预测aSAH后迟发性脑缺血的曲线下面积(AUC)为0.862(95%CI:0.781~0.942),敏感度为76.50%,特异度为83.82%,截断值为1.19 ng/L;NeuN预测aSAH后迟发性脑缺血的AUC为0.771(95%CI:0.672~0.870),敏感度为78.10%,特异度为76.50%,截断值为0.47μg/mL;AQP4联合NeuN预测aSAH后迟发性脑缺血的AUC为0.879(95%CI:0.811~0.948),敏感度为90.60%,特异度为75.00%;联合诊断的AUC值高于NeuN单独诊断(Z=2.476,P=0.013)。结论aSAH后迟发性脑缺血患者血清NeuN水平下调,AQP4水平上调,AQP4联合NeuN可作为预测aSAH后迟发性脑缺血的评估指标。

精子核蛋白组型转换在预测复发性流产中的价值

目的 探讨精子核蛋白组型转换在复发性流产中的预测价值。方法 选取2019—2022年在本院生殖医学门诊就诊的生育检测指标完整的不孕症夫妇521对,其中男性年龄23~56岁。排除导致复发性流产的因素,包括女性的年龄、体质指数、代谢性疾病、抗磷脂综合征、子宫及附件异常、剖宫产史、宫内肌瘤/宫颈锥切等手术史、男女外周血染色体异常、吸烟/酗酒等不良生活史。根据流产情况分为复发性流产组(自然流产≥2次)、自然流产1次组和未流产组,应用GraphPad6.0统计软件进行Tukey多重比较各组精子核蛋白组型转换的差异;采用Pearson法分析精子核蛋白组型转换与复发性流产的相关性;采用敏感性、特异性、阳性预测值、阴性预测值、Youden指数和比值比分析精子核蛋白组型转换对复发性流产的预测价值。结果 复发性流产组(42例)的精子核蛋白组型转换异常率[(33.31±13.83)%]显著高于未流产组[(26.85±15.38)%]和流产1次组[(28.20±12.50)%,P<0.05]。Pearson相关分析结果显示,精子核蛋白组型转换异常率与复发性流产呈显著正相关。精子核蛋白组型转换预测复发性流产的敏感性为45.24%、特异性为73.64%。阳性预测值(15.70%)、阴性预测值(92.53%)、Youden指数(18.88%)及比值比(2.31)均高于高DNA可染色性(HDS)的阳性预测值(10.64%)、阴性预测值(90.31%)、Youden指数(1.05%)及比值比(1.11)。结论 在复发性流产患者的配偶中,精子核蛋白不成熟度显著升高,且其与复发性流产呈显著正相关。同时精子核蛋白组型转换异常可能是复发性流产的重要男性因素,其对临床中复发性流产的预测价值较高。

布尼亚病毒核蛋白的结构与功能

布尼亚病毒分布广,传染性强,致死率高,是对全世界的公共卫生事业有重大影响的负链RNA病毒,开发疫苗和寻找药物是预防布尼亚病毒感染的关键。病毒的核蛋白(nucleoprotein,NP)是合成病毒RNA所必需的,其与病毒RNA结合形成核衣壳,参与病毒的组装与RNA转录,在病毒的增殖过程中发挥重要作用;此外,NP还具有B细胞和T细胞表位,能诱发细胞免疫和体液免疫,因此,NP是疫苗设计和药物开发的理想靶点。鉴于NP的丰度和特异性,NP也常用于病毒病的检测。目前,越来越多的布尼亚病毒NP结构及NP-RNA复合物的结构得以解析,研究者已经通过这些结构发现2个重要的抗病毒靶点,即NP的末端臂和RNA结合口袋。本文就布尼亚病毒NP的功能和三维结构以及NP作为抗病毒靶点的研究进展作一综述,以期为布尼亚病毒病的防治提供理论依据。

表柔比星联合nab-紫杉醇序贯治疗三阴性乳腺癌的临床疗效

目的探讨表柔比星联合nab-紫杉醇序贯治疗三阴性乳腺癌的临床疗效。方法选择2015年1月至2020年1月四川绵阳四〇四医院收治的87例三阴性乳腺癌患者作为研究对象,根据治疗方法不同分为研究组(采用表柔比星、nab-紫杉醇序贯化疗,45例)和对照组(采用表柔比星、nab-紫杉醇联合化疗,42例)。比较两组治疗24周后的疗效,治疗前、治疗24周后血清癌症预后因子[血管内皮生长因子(VEGF)亚型]、血清肿瘤标志物[糖类抗原(CA)125、CA15-3]水平、免疫组织化学指标[核蛋白Ki-67、程序性细胞死亡配体1(PD-L1)、细胞间黏附分子-1(ICAM-1)]及脏器损伤相关指标[肌酸激酶(CK)、肌酸激酶同工酶(CK-MB)、γ-谷胺酰转肽酶(γ-GT)]水平;比较两组治疗期间不良反应发生率、治疗后随访6个月的转移率、复发率、死亡率。结果治疗24周后,研究组总有效率显著高于对照组[62.22%(28/45)比40.47%(17/42)](P<0.05),研究组疗效优于对照组(Z=1.963,P=0.050)。治疗24周后,两组患者血清VEGF-A、VEGF-B、CA125、CA15-3、Ki-67、PD-L1、ICAM-1水平均较治疗前降低,CK、CK-MB、γ-GT水平均较治疗前升高(P<0.05),且治疗后研究组上述各指标水平均低于对照组(均P<0.01)。治疗后,两组骨髓抑制、恶心呕吐、脱发、肝肾功能异常、腹泻发生率比较差异无统计学意义(P>0.05),研究组总不良反应发生率显著低于对照组[20.00%(9/45)比45.24%(19/42)](P<0.05)。治疗后随访6个月,研究组失访2例,有效病例43例;对照组失访2例,有效病例40例。治疗后6个月,研究组转移率显著低于对照组[4.65%(2/43)比20.00%(8/40)](P<0.05),而两组复发率、死亡率比较差异无统计学意义(均P>0.05)。结论表柔比星联合nab-紫杉醇序贯治疗可以有效降低三阴性乳腺癌患者血清肿瘤标志物、免疫组织化学指标的表达,且安全性较好。

新型冠状病毒结构蛋白单克隆抗体的制备与鉴定

目的 制备针对新型冠状病毒(SARS-CoV-2)刺突蛋白(S蛋白)S1亚基、核衣壳蛋白(N蛋白)的小鼠源性单克隆抗体(mAbs),进行功能和应用的初步鉴定。制备具备中和活性的高亲和力人鼠嵌合抗体。方法 用重组表达的SARS-CoV-2 S蛋白S1亚基和N蛋白分别免疫BALB/c小鼠,利用常规B细胞杂交瘤技术制备抗S蛋白S1亚基mAbs和抗N蛋白mAbs,并鉴定其在Western blotting、免疫组织化学染色中的应用价值;建立检测可溶性S蛋白和N蛋白的夹心ELISA方法;基于假病毒-荧光素酶检测系统,检测特异性抗体的中和活性。将分泌具备中和活性抗S蛋白S1亚基mAbs的杂交瘤细胞株测序,制备人鼠嵌合抗体。结果 获得53株分泌小鼠抗SARS-CoV-2 S蛋白S1亚基mAbs的杂交瘤细胞株,克隆号分别命名XA326.1~XA326.53;获得42株分泌小鼠抗SARS-CoV-2 N蛋白mAbs的杂交瘤细胞株,克隆号分别命名XA327.1~XA327.42。初步鉴定出9株可用于Western blotting检测可溶性SARS-CoV-2 S蛋白S1亚基的mAbs、8株可用于Western blotting检测可溶性SARS-CoV-2 N蛋白的mAbs;9株可用于免疫组织化学染色的抗SARS-CoV-2 S蛋白S1亚基mAbs、4株可用于免疫组织化学染色的抗SARS-CoV-2 N蛋白mAbs。构建以mAb XA326.7为包被抗体、生物素标记的mAb XA326.19为检测抗体的检测可溶性SARS-CoV-2 S蛋白S1亚基的夹心ELISA试剂盒;采用mAb XA327.37作为包被抗体、生物素标记的mAb XA327.25为检测抗体,可以建立检测可溶性SARS-CoV-2 N蛋白的夹心ELISA试剂盒。XA326.4、XA326.7、XA326.8、XA326.49 mAbs具有中和活性,可以在假病毒感染实验中阻断病毒对人ACE2阳性细胞的感染。结论 成功制备出一套小鼠抗SARS-CoV-2 S蛋白S1亚基、N蛋白的mAbs,可用于多种免疫学检测技术,XA326.4、XA326.7、XA326.8、XA326.49中和活性抗体具有潜在的临床应用价值。

肝病自身抗体对原发性胆汁性胆管炎的临床价值

目的探讨肝病自身抗体抗线粒体抗体(AMA)、抗线粒体抗体-M2型(AMA-M2)、抗gp210抗体、抗sp100抗体对原发性胆汁性胆管炎(PBC)的诊断价值及预后价值。方法回顾性分析2020年3月至2022年2月于福建医科大学附属泉州第一医院就诊的88例PBC患者、100例非PBC肝功能不全者和140名健康对照者的临床资料,计算AMA、AMA-M2、抗gp210抗体和抗sp100抗体诊断PBC的灵敏度、特异度、AUC曲线下面积,并进一步比较各抗体阳性组和阴性组PBC患者肝脏酶学指标丙氨酸氨基转移酶(ALT)、天门冬氨酸氨基转移酶(AST)、碱性磷酸酶(ALP)和γ-谷氨酰转移酶(γ-GGT)和免疫球蛋白IgG、IgM水平差异是否显著。结果AMA、AMA-M2、抗gp210抗体、抗sp100抗体诊断PBC的灵敏度分别为70.45%、77.27%、39.77%和17.05%,特异度分别为90.42%、92.08%、95.42%、96.67%,AUC曲线下面积分别为0.8044,0.8468,0.6759,0.5686。PBC患者中AMA、AMA-M2阳性组与阴性组间AST、ALT、ALP、GGT、IgG和IgM水平差异无统计学意义(P>0.05),而抗gp210抗体、抗sp100抗体阳性组ALP、GGT和IgM水平高于阴性组,差异有统计学意义(P<0.05)。抗gp210抗体在肝硬化组与非肝硬化组的阳性率分别为60.87%(14/23)和32.31%(21/65),差异有统计学意义(P<0.05)。结论AMA、AMA-M2、抗gp210抗体、抗sp100抗体对诊断PBC均具有较高的特异度,但敏感度不一,其中AMA-M2对PBC的诊断效能较好;抗gp210抗体、抗sp100抗体与PBC疾病活动度有关,对PBC具有一定的预测价值。