Anti-PD1 antibody and not anti-LAG-3 antibody improves the antitumor effect of photodynamic therapy for treating metastatic breast cancer

Photodynamic therapy(PDT)has limited effects in treating metastatic breast cancer.Immune checkpoints can deplete the function of immune cells;however,the expression of immune checkpoints after PDT is unclear.This study investigates whether the limited e±cacy of PDT is due to upregulated immune checkpoints and tries to combine the PDT and immune checkpoint inhibitor to observe the e±cacy.A metastatic breast cancer model was treated by PDT mediated by hematoporphyrin derivatives(HpD-PDT).The anti-tumor effect of HpD-PDT was observed,as well as CD4þT,CD8þT and calreticulin(CRT)by immunohistochemistry and immunofluorescence.Immune checkpoints on T cells were analyzed byflow cytometry after HpD-PDT.When combining PDT with immune checkpoint inhibitors,the antitumor effect and immune effect were assessed.For HpD-PDT at 100 mW/cm2 and 40,60 and 80 J/cm2,primary tumors were suppressed and CD4þT,CD8þT and CRT were elevated;however,distant tumors couldn't be inhibited and survival could not be prolonged.Immune checkpoints on T cells,especially PD1 and LAG-3 after HpD-PDT,were upregulated,which may explain the reason for the limited HpD-PDT effect.After PDT combined with anti-PD1 antibody,but not with anti-LAG-3 antibody,both the primary and distant tumors were signi-cantly inhibited and the survival time was prolonged,additionally,CD4þT,CD8þT,IFN-þCD4þT and TNF-þCD4þT cells were signi-cantly increased compared with HpD-PDT.HpD-PDT could not combat metastatic breast cancer.PD1 and LAG-3 were upregulated after HpD-PDT.Anti-PD1 antibody,but not anti-LAG-3 antibody,could augment the antitumor effect of HpD-PDT for treating metastatic breast cancer.

Atypical Guillain-Barrésyndrome with positive anti-sulfatide,anti-GT1b,and anti-GT1a antibodies:A case report

BACKGROUND The role of diverse antibodies in mediating peripheral nerve injury in Guillain-Barrésyndrome(GBS)is becoming clearer,but positivity for multiple antibodies in one case is uncommon.To our knowledge,this is the first case involving GBS with positive anti-sulfatide,anti-GT1a,and anti-GT1b antibodies.CASE SUMMARY A 20-year-old female patient was admitted to the hospital due to weakness of limbs for 5 d,and deterioration of the weakness and muscle aches for 1 d.The patient's limbs were weak,but the tendon reflexes in the part of the limbs were normal.There was no comorbid peripheral nociception or deep sensory dysfunction.She was diagnosed with GBS and was discharged after receiving intravenous human immunoglobulin pulse therapy.CONCLUSION In this article,the clinical manifestations,neurophysiological examination,and auxiliary examination findings of a GBS patient positive for multiple antibodies were analyzed to improve the identification of the disease by clinical physicians at an early stage.

Miller fisher syndrome with positive anti-GQ1b/GT1a antibodies associated with COVID-19 infection:A case report

BACKGROUND Miller fisher syndrome(MFS)is a variant of Guillain-Barrésyndrome,an acute immune-mediated peripheral neuropathy that is often secondary to viral infections.Anti-ganglioside antibodies play crucial roles in the development of MFS.The positive rate of ganglioside antibodies is exceptionally high in MFS patients,particularly for anti-GQ1b antibodies.However,the presence of other ganglioside antibodies does not exclude MFS.CASE SUMMARY We present a 56-year-old female patient who suddenly developed right blepharoptosis and progressively worsening vision in both eyes.There were flu symptoms prior to onset,and a coronavirus disease 2019 test was positive.On physical examination,the patient exhibited bilateral extraocular muscle paralysis,weakened reflexes in both limbs,and impaired coordination.The cerebrospinal fluid examination results showed no obvious abnormalities.Bilateral peroneal nerve F-waves were not extracted.Serum anti-GD1b IgG and anti-GT1a IgG antibodies were positive.The patient received intravenous methylprednisolone(1000 mg/day),with the dosage gradually decreased.Additionally,intravenous high-dose immunoglobulin treatment was administered for 5 days(0.4 g/kg/day)from day 2 to day 6 of hospitalization.The patient’s symptoms improved after treatment with immunoglobulins and hormones.CONCLUSION Positive ganglioside antibodies may be used as supporting evidence for the diagnosis;however,the diagnosis of MFS is more reliant on clinical symptoms.

Diagnostic significance of serum levels of serum amyloid A,procalcitonin,and high-mobility group box 1 in identifying necrotising enterocolitis in newborns

BACKGROUND Necrotising enterocolitis(NEC)is a critical gastrointestinal emergency affecting premature and low-birth-weight neonates.Serum amyloid A(SAA),procalcitonin(PCT),and high-mobility group box 1(HMGB1)have emerged as potential biomarkers for NEC due to their roles in inflammatory response,tissue damage,and immune regulation.AIM To evaluate the diagnostic value of SAA,PCT,and HMGB1 in the context of NEC in newborns.METHODS The study retrospectively analysed the clinical data of 48 newborns diagnosed with NEC and 50 healthy newborns admitted to the hospital.Clinical,radiological,and laboratory findings,including serum SAA,PCT,and HMGB1 Levels,were collected,and specific detection methods were used.The diagnostic value of the biomarkers was evaluated through statistical analysis,which was performed using chi-square test,t-test,correlation analysis,and receiver operating characteristic(ROC)analysis.RESULTS The study demonstrated significantly elevated levels of serum SAA,PCT,and HMGB1 Levels in newborns diagnosed with NEC compared with healthy controls.The correlation analysis indicated strong positive correlations among serum SAA,PCT,and HMGB1 Levels and the presence of NEC.ROC analysis revealed promising sensitivity and specificity for serum SAA,PCT,and HMGB1 Levels as potential diagnostic markers.The combined model of the three biomarkers demonstrating an extremely high area under the curve(0.908).CONCLUSION The diagnostic value of serum SAA,PCT,and HMGB1 Levels in NEC was highlighted.These biomarkers potentially improve the early detection,risk stratification,and clinical management of critical conditions.The findings suggest that these biomarkers may aid in timely intervention and the enhancement of outcomes for neonates affected by NEC.

Detection of Serum Antibodies of Porcine Pseudorabies Virus(PRV)in Binzhou City in 2014

To understand immunization effect of pseudorabies vaccine and infection status of porcine pseudorabies （PR） in pig farms and guide prevention and control against PR, 453 copies of blood samples collected from 43 different scale pig farms in four counties （districts） of Binzhou City were detected with ELISA to investigate PRV gB antibody and gE antibody. Detection results showed that the gB antibody positive rate of sows was 75.58%, and that of fattening pigs was 68.67% ; the pig farms with positive rate higher than 70% accounted for 74.42% of total survey pig farms. The PRV gE antibody positive rates of sows and fatte- ning pigs were 25.41% and 26.67%, and the positive rates of pig farms were 46.51% and 44.33%, respectively. There were regional differences among counties （districts）.

Emergent Serum Therapy and Antibody Medicine to Counteract Sudden Attacks of COVID-19 and Other Pathogenic Epidemics

The science behind two life-saving protocols to combat COVID-19 and future pathogenic epidemics is presented for immediate implementation.

Treadmill exercise in combination with acousto-optic and olfactory stimulation improves cognitive function in APP/PS1 mice through the brain-derived neurotrophic factor-and Cygb-associated signaling pathways

A reduction in adult neurogenesis is associated with behavioral abnormalities in patients with Alzheimer's disease.Consequently,enhancing adult neurogenesis represents a promising therapeutic approach for mitigating disease symptoms and progression.Nonetheless,nonpharmacological interventions aimed at inducing adult neurogenesis are currently limited.Although individual non-pharmacological interventions,such as aerobic exercise,acousto-optic stimulation,and olfactory stimulation,have shown limited capacity to improve neurogenesis and cognitive function in patients with Alzheimer's disease,the therapeutic effect of a strategy that combines these interventions has not been fully explored.In this study,we observed an age-dependent decrease in adult neurogenesis and a concurrent increase in amyloid-beta accumulation in the hippocampus of amyloid precursor protein/presenilin 1 mice aged 2-8 months.Amyloid deposition became evident at 4 months,while neurogenesis declined by 6 months,further deteriorating as the disease progressed.However,following a 4-week multifactor stimulation protocol,which encompassed treadmill running(46 min/d,10 m/min,6 days per week),40 Hz acousto-optic stimulation(1 hour/day,6 days/week),and olfactory stimulation(1 hour/day,6 days/week),we found a significant increase in the number of newborn cells(5'-bromo-2'-deoxyuridine-positive cells),immature neurons(doublecortin-positive cells),newborn immature neurons(5'-bromo-2'-deoxyuridine-positive/doublecortin-positive cells),and newborn astrocytes(5'-bromo-2'-deoxyuridine-positive/glial fibrillary acidic protein-positive cells).Additionally,the amyloid-beta load in the hippocampus decreased.These findings suggest that multifactor stimulation can enhance adult hippocampal neurogenesis and mitigate amyloid-beta neuropathology in amyloid precursor protein/presenilin 1 mice.Furthermore,cognitive abilities were improved,and depressive symptoms were alleviated in amyloid precursor protein/presenilin 1 mice following multifactor stimulation,as evidenced by Morris water maze,novel object recognition,forced swimming test,and tail suspension test results.Notably,the efficacy of multifactor stimulation in consolidating immature neurons persisted for at least 2weeks after treatment cessation.At the molecular level,multifactor stimulation upregulated the expression of neuron-related proteins(NeuN,doublecortin,postsynaptic density protein-95,and synaptophysin),anti-apoptosis-related proteins(Bcl-2 and PARP),and an autophagyassociated protein(LC3B),while decreasing the expression of apoptosis-related proteins(BAX and caspase-9),in the hippocampus of amyloid precursor protein/presenilin 1 mice.These observations might be attributable to both the brain-derived neurotrophic factor-mediated signaling pathway and antioxidant pathways.Furthermore,serum metabolomics analysis indicated that multifactor stimulation regulated differentially expressed metabolites associated with cell apoptosis,oxidative damage,and cognition.Collectively,these findings suggest that multifactor stimulation is a novel non-invasive approach for the prevention and treatment of Alzheimer's disease.

Analysis of Specific Th1/Th2 Helper Cell Responses and IgG Subtype Antibodies in Anti-CD4 Monoclonal Antibody Treated Mice with Autoimmune Cardiomyopathy

The cytokine repertoire of ADP/ATP carrier-specific humoral immune responses and the cytokine-dependent anti-ADP/ATP carrier antibody IgG subclasses were examined in a cohort of ADP/ATP carrier-immunized BALB/c mice treated with anti-CD4 monoclonal antibody. Eighteen male BALB/c mice （6–8 weeks old） were randomized into 3 groups： dilated cardiomyopathy （DCM） group, DCM-tolerance （Tol） group and control group. The mice in DCM group were immunized with the peptides derived from human ADP/ATP carrier protein for 6 months and mice in the control group were sham-immunized, while the mice in DCM-Tol group were immunized with ADP/ATP carrier protein and anti-CD4 McAb simultaneously. Serum autoantibody against ADP/ATP carrier and IgG subclasses were measured by ELISA, intracellular cytokines IFN-γ and IL-4 of Th cells were moni- tored with flow cytometry, and splenic T cell cytokines IFN-γ, IL-2, IL-4 and IL-6 were detected by using real-time fluorescent quantitative PCR. The results showed that the autoantibody against ADP/ATP carrier was found in all mice in DCM group, and the antibody level, serum IgG1 and IgG2a subclasses, cytokines in T cells and Th cells were all elevated in DCM group, as compared with those in control group （P〈0.01）. On the other hand, in DCM-Tol group, the autoantibody level and contents of all the cytokines were significantly different from those in DCM group （P〈0.01）, and were close to those in control group. And the levels of IgG1, IgG2a, IgG2b and IgG3 were influenced, to varying degrees, by anti-CD4 McAb as compared with those in DCM group. All these four types of IgG subclasses were substantially decreased in DCM-Tol group as compared with DCM group. It is concluded that the treatment with anti-CD4 McAb could prevent the activation of T cells, reverse the abnormal secretion of cytokines and the imbalance between Th1/Th2 cell subsets and abnormal production of autoantibody against ADP/ATP carrier, and eventually avoid myocardial injuries.

Analysis of Detecting HIV-1 Antibody in Paired Urine and Serum Specimens from Drug Users by ELISA

Objective- To compare the consistency of the results from detecting HIV-1 antibody in the paired urine and serum specimens from drug users by ELISA. Methods: The paired urine and serum specimens from 273 drug users detained at a detoxification unit were collected, and the HIV-1 antibodies in the specimens of them were screened by urine and serum ELISA kits, respectively. Results: Of 273 serum specimens, 94 ones showed positive reaction and among 94 counterpart urine specimens, 93 ones also appeared positive reaction. Taking the results together,the consistent rate of HIV-1 antibody screened by urine and serum ELISA kits was 99.6%. Conclusion: The urine ELISA kit, which screened HIV-1 antibody of urine showing almost the same results tested by serum ELISA kit, is reliable. It is proposed that urine ELISA be introduced in many fields.

Generation of Antibodies Against DMRT1 and DMRT4 of Oreochromis aurea and Analysis of Their Expression Profile in Oreochromis aurea Tissues

Sex determination is composed of somatic and germ-line sex differentiation hierarchies whose interaction is poorly understood. A single gene known to control somatic sex determination, the DM-domain containing （Doublesex/Mab-3 DNA-binding motif） gene, is highly conserved across species. Vertebrate DMRT1 （DM-related transcription factor 1） expression occurs predominantly in the testis. Here, however, isolated two distinct DM-domain cDNA from Oreochromis aurea ovary and testis have been named DMRT4 （DM-related transcription factor 4） and DMRT1 by BLAST, respectively. Despite high homology in the DM-domain there is little similarity outside the DM-domain.To better understand the structure, function, and possible roles of DMRT4 and DMRT1 as potential candidates for sex differentiation and sex determination, the intact regions encoding DMRT4 and DMRT1 obtained by PCR were sub-cloned into the vector pMAL-c2x and introduced into the Escherichia coli TB1 cell for efficient fusion expression. After purification and cleavage, DMRT4 and DMRT1 proteins were used to immunize adult rabbits following standard protocols. Consequently, it was found by using Western blot analysis that polyclonal antibodies against DMRT4 and DMRT1 had high specificity. The relative expression levels of DMRT4 and DMRT1 mRNA were determined by fluorescent Real-time RT-PCR in female and male Oreochromis aurea with 13-actin as the internal standard. DMRT1 was expressed only in testis, whereas DMRT4 was over expressed in the ovary, but in both female and male, a slight expression in the brain was also detected. Statistical analysis showed that in the brain, mean DMRT4 mRNA levels in female were significantly higher than in male. Meanwhile, the expression of DMRT4 and DMRT1 protein was also analyzed using the purified antibodies through Western blot and immunohistochemistry. It was found that DMRT4 was exclusively expressed in the ovary and DMRT1 in the testis. Study on DMRT4 and DMRT1 expression facilitated the elucidation of their roles and the understanding of sex differentiation of fish.

Establishment of Monoclonal Antibody Competitive ELISA Using Monoclonal Antibody Against VP1 Protein of Asia 1 Type Foot-and-Mouth Disease Virus

Using the purified VP1 protein of Asia 1 type foot-and-mouth disease virus as the antigen, the purified monoclonal antibody was labeled by the sodium periodate method and the monoclonal antibody competitive ELISA was established in this study. Ten positive porcine foot-and-mouth disease serums and more than two hundreds negative serum were tested, and the results were the same as the background of samples. The sensitivity test and replicate test indicated that this method was stable and sensitive, which was suitable for monitoring Asia 1 type porcine foot-and-mouth disease virus antibody.

2017—2021年云南省猪瘟病原学和血清学监测情况分析

为全面了解2017—2021年云南省猪瘟(CSF)流行及免疫状况,预估疫病流行和发生风险,采集病原学样品63 224份、血清学样品146 547份,进行CSF病原和血清抗体监测。结果显示:2017—2021年云南省CSF免疫抗体合格率分别为82.94%、81.47%、80.79%、80.24%、82.53%,规模场合格率略高于散养户,但差异不显著(P>0.05);CSF病原核酸阳性率分别为0.52%、0.74%、1.10%、1.02%、2.42%,散养户、屠宰场阳性率高于规模场。结果表明:自2017年CSF退出强制免疫后,云南省CSF免疫抗体合格率均达到国家标准,但病原阳性率整体呈上升趋势,且分布范围广,免疫密度低和抗体合格率低的散养户发生CSF的风险较高,提示应对其加强监测和免疫。

胸腺肽α_1单克隆抗体的制备及其鉴定

目的 研制胸腺肽α1(Tα1)单克隆抗体 ,为Tα1作用机制的阐明奠定基础。方法 用戊二醛将Tα1和牛血清白蛋白进行交联 ,以其交联物为抗原 ,按常规淋巴细胞杂交瘤技术制备单克隆抗体 ,并采用间接酶联免疫吸附 (ELISA)法和Western免疫印迹来鉴定单克隆抗体的特异性。结果 建立了分泌抗Tα1抗体的杂交瘤细胞株 ,能特异性地与Tα1结合 ,但与单纯牛血清白蛋白、酵母抽提物和转染Tα1酵母未诱导表达产物无反应。其抗体效价为 1∶10 2 4 0 ,Ig亚类为IgG2b。结论 本研究建立了针对Tα1高效价、高特异性单克隆抗体细胞株。

老年急性脑梗死患者血清Del-1和IL-17水平变化及与梗死面积和预后的关系

目的 探究老年急性脑梗死(ACI)患者血清内皮发育调节基因-1(Del-1)、白细胞介素-17(IL-17)水平与脑梗死面积、病情严重程度及预后的关系。方法 回顾性收集2019-01-2021-12厦门大学附属东南医院收治的126例老年ACI患者(病例组)及105名健康体检者(对照组)的临床资料,根据脑梗死面积将病例组患者分为大面积梗死组(梗死最大直径>5 cm,n=14)、中面积梗死组(梗死最大直径3~5 cm,n=53)和小面积梗死组(梗死最大直径<3 cm,n=59);根据NIHSS评分分为重症组(NIHSS评分≥16分,n=13)、中症组(5分<NIHSS评分<15分,n=58)和轻症组(NIHSS评分≤5分,n=55);根据m RS评分分为预后良好组(mRS评分≤2分,n=81)和预后不良组(mRS评分>2分,n=45)。采用荧光免疫法检测血清Del-1、IL-17水平,受试者工作特征(ROC)曲线评估血清Del-1、IL-17水平预测老年ACI患者预后的价值。结果 病例组吸烟史比例、收缩压、舒张压、糖化血红蛋白、血清IL-17水平均高于对照组,血清Del-1水平低于对照组(P<0.05)。不同脑梗死面积患者血清Del-1水平比较,小面积梗死组>中面积梗死组>大面积梗死组;血清IL-17水平比较,小面积梗死组<中面积梗死组<大面积梗死组(P<0.05)。轻症组血清Del-1水平高于中症组、重症组(P<0.05),不同病情严重程度患者血清IL-17水平比较,重症组>中症组>轻症组(P<0.05)。预后良好组患者血清Del-1水平高于预后不良组,血清IL-17水平低于预后不良组(P<0.05)。ROC曲线分析显示,血清Del-1、IL-17预测ACI患者预后的AUC值分别为0.763、0.747(P<0.05)。结论 老年ACI患者血清Del-1水平较低,IL-17水平较高,二者与患者脑梗死面积、病情严重程度和预后关系密切,可作为预测老年ACI患者预后的可靠指标。

PD-1单抗后线辅助治疗晚期结直肠癌患者对胃肠功能、肿瘤标志物的影响

目的分析程序性死亡受体-1(PD-1)单抗后线辅助治疗晚期结直肠癌患者对胃肠功能、肿瘤标志物的影响。方法前瞻性选取2020年1月至2023年1月海口市人民医院收治的105例晚期结直肠癌患者为研究对象,按照随机数字表法将患者分为对照组(n=50)、研究组(n=55)。对照组采用阿帕替尼治疗,研究组在对照组的基础上联合PD-1单抗后线治疗,3周为1个疗程,共治疗4个疗程。比较两组的临床疗效、胃肠功能改善时间,治疗前、治疗12周后的肿瘤标志物[癌胚抗原、糖类抗原(CA)72-4、CA199]、免疫功能(CD4^(+)、CD8^(+)、CD4^(+)/CD8^(+))指标以及不良反应发生情况。结果治疗12周后,研究组的客观缓解率(ORR)、病情控制率(DCR)分别为78.33%、90.91%,均高于对照组(58.00%、70.00%),差异均有统计学意义(P<0.05)。研究组患者的首次排便时间、首次排气时间、肠鸣音消退时间分别为(51.50±5.02)、(35.52±4.25)、(44.28±4.80)d,均短于对照组[(69.37±6.40)、(50.63±5.10)、(53.50±6.15)d],差异均有统计学意义(P<0.05)。治疗12周后,研究组癌胚抗原、CA72-4、CA199水平分别为(12.57±1.64)ng/mL、(17.60±3.45)U/mL、(58.29±8.70)kU/L,均低于对照组[(17.24±2.50)ng/mL、(27.59±4.67)U/mL、(82.65±12.29)kU/L],差异均有统计学意义(P<0.05)。治疗12周后,研究组CD4^(+)、CD4^(+)/CD8^(+)水平分别为(21.42±3.54)%、0.73±0.30,均低于对照组[(28.39±4.38)%、1.12±0.41],CD8^(+)水平为(30.36±4.52)%,高于对照组[(25.71±3.30)%],差异均有统计学意义(P<0.05)。研究组的不良反应发生率为9.09%,稍低于对照组(16.00%),但两组比较差异无统计学意义(P>0.05)。结论通过PD-1单抗后线辅助治疗可有效改善晚期结直肠癌患者的临床疗效,效果明显,改善胃肠功能,降低肿瘤标志物指标水平,调节机体免疫功能,可为临床干预此病提供参考。

扶正驱邪方联合新辅助化疗对三阴性乳腺癌患者肿瘤复发、血清TK1 水平及免疫功能的影响

【目的】探究扶正驱邪方联合新辅助化疗对三阴性乳腺癌(TNBC)患者肿瘤复发、血清胸苷激酶1(TK1)水平及免疫功能的影响。【方法】将80例TNBC气阴两虚型患者随机分为联合组和对照组,每组各40例。对照组给予AC-T序贯化疗方案(多柔比星与环磷酰胺联合并序贯多西他赛)治疗,联合组在对照组的基础上加用扶正驱邪方治疗。1个疗程为21 d,连续治疗4个疗程。观察2组患者治疗前后中医证候积分、生活质量Karnofsky功能状态(KPS)评分、肿瘤标志物[糖类抗原125(CA125)、糖类抗原153(CA153)、TK1]水平及T淋巴细胞亚群的变化情况,比较2组患者的临床疗效以及肿瘤的转移、复发情况。【结果】(1)治疗4个疗程后,联合组的总有效率为87.50%(35/40),对照组为67.50%(27/40),组间比较(χ2检验),联合组的疗效明显优于对照组(P<0.05)。(2)治疗后,2组患者的中医证候积分均较治疗前明显降低(P<0.05),KPS评分均较治疗前明显升高(P<0.05),且联合组对中医证候积分的降低幅度及对KPS评分的升高幅度均明显优于对照组(P<0.05或P<0.01)。(3)治疗后,2组患者的血清CA125、CA153、TK1水平均较治疗前明显降低(P<0.05),且联合组对血清CA125、CA153、TK1水平的降低幅度均明显优于对照组(P<0.01)。(4)治疗后,2组患者的T细胞CD3+、CD4+水平和CD4+/CD8+比值均较治疗前明显升高(P<0.05),CD8+水平均较治疗前明显降低(P<0.05),且联合组对T细胞CD3+、CD4+水平和CD4+/CD8+比值的升高幅度及对CD8+水平的降低幅度均明显优于对照组(P<0.05或P<0.01)。(5)经过1年的随访调查,联合组的肿瘤复发率和肿瘤转移率分别为7.50%(3/40)和12.50%(5/40),明显低于对照组的25.00%(10/40)和35.00%(14/40),组间比较,差异均有统计学意义(P<0.05)。【结论】扶正驱邪方联合新辅助化疗对于TNBC气阴两虚型患者的治疗效果较好,能有效改善患者的免疫功能,降低血清肿瘤标志物水平,提高患者的生活质量,降低肿瘤复发和转移的发生率。

H5N1亚型禽流感病毒神经氨酸酶蛋白在昆虫细胞中的表达与鉴定

研究旨在利用昆虫细胞-杆状病毒表达系统制备H5N1亚型禽流感病毒(AIV)的神经氨酸酶(NA)蛋白。试验为获得具有良好反应原性的NA蛋白,将H5N1亚型AIV(DK/YN/S4469/2022)密码子优化后的N1基因插入载体pFastBac1,构建的重组转移载体pFastBac1-N1转化至DH10Bac感受态细胞,经过蓝白斑筛选获得阳性重组杆粒rBacmid-N1,然后将重组杆粒转染至sf9昆虫细胞,获得N1亚型重组杆状病毒。结果显示:N1重组蛋白在sf9昆虫细胞中正确表达,并且该重组蛋白具有良好的反应原性,能够与N1蛋白单克隆抗体3F3发生特异性反应,其仅能识别N1亚型鸡血清,特异性较好。研究成功制备了N1重组蛋白,为NA蛋白表达方法提供参考并为临床血清样品的N1抗体检测方法的建立奠定基础。

血清中PD-L1含量与胃癌病人预后的关系

目的:分析胃癌病人血清中程序性死亡配体1(programmed death-ligand 1,PD-L1)含量及其与肿瘤分期和预后的关系。方法:收集我院2018年8月至2019年12月60例胃癌病人血清,15例健康成人血清作为对照,通过酶联免疫吸附试验(ELISA)检测血清PD-L1的含量,比较胃癌病人与健康成人血清PD-L1含量的差异,分析胃癌病人血清PD-L1的含量与病期及预后的关系。免疫组织化学检测肿瘤组织内PD-L1的表达情况,并分析其与血清中PD-L1含量的相关性。结果:胃癌组血清中PD-L1的含量明显高于对照组。Ⅲ、Ⅳ期胃癌病人血清PD-L1含量明显高于Ⅰ、Ⅱ期胃癌病人。胃癌病人合并腹膜转移者血清PD-L1含量明显升高,提示预后不良。生存分析显示,血清PD-L1含量高的胃癌病人生存期明显短于低者。检测肿瘤组织标本中PD-L1的表达水平,发现其与血清中PD-L1含量无明显相关性。结论:胃癌病人血清中PD-L1含量高于健康成人。随病期进展逐渐升高。胃癌伴腹膜转移的病人血清中PD-L1升高明显。血清中PD-L1含量与肿瘤组织中的PD-L1表达不相关。

抗单纯疱疹病毒1型的IgY制备及其生物学活性检测

目的:制备抗单纯疱疹病毒1型(HSV-1)的卵黄抗体(IgY),探讨该抗体的生物学活性,阐明其抗HSV-1的能力。方法:制备HSV-1灭活疫苗,采用鸡胸多点注射法免疫高产蛋鸡,采用聚乙二醇-6000(PEG-6000)法提纯IgY,采用间接ELISA法、十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)技术和蛋白浓度定量试剂盒测定IgY效价、纯度及蛋白水平,采用间接ELISA法检测IgY中和病毒的能力,测定病毒阻断率,以Vero细胞为病毒感染靶细胞,测定不同抗体浓度(0.01560、0.03125、0.06250、0.12500、0.50000、和1.00000 g·L^(-1))作用后细胞病变效应(CPE),根据CPE程度测定IgY体外抗HSV-1能力。结果:制备的IgY效价随免疫时间逐渐升高,达到1/1024000后保持稳定;IgY轻链重链条带清晰;IgY平均蛋白水平为11.544 g·L^(-1);IgY对HSV-1的阻断率可高达77.90%;HSV-1致CPE程度随抗体浓度升高而逐渐降低,当IgY浓度达到0.50000g·L^(-1)及以上时,25%以下细胞(甚至无细胞)发生病变。结论:成功制备出抗HSV-1的IgY,该抗体对HSV-1具有明显的中和阻断能力和体外抑制活性,可用于药物及诊断方法的开发。

老年脑梗死患者血清ACA、抗β2GP1、Hcy和hs-CRP水平变化及其与疾病严重程度的关系

目的:探讨老年脑梗死患者血清抗心磷脂抗体(ACA)、抗β2糖蛋白1抗体(抗β2GP1)、同型半胱氨酸(Hcy)和超敏C反应蛋白(hs-CRP)水平变化及其与疾病严重程度的关系。方法:选取148例老年脑梗死患者为观察组,根据神经功能缺损(NDS)评分分为轻度组(n=30)、中度组(n=32)及重度组(n=86);另选同期76名体检健康老年人为对照组。比较观察组和对照组及观察组不同病情程度患者的血清ACA、抗β2GP1、Hcy、hs-CRP水平,分析其与疾病严重程度的关系。结果:观察组患者血清ACA、抗β2GP1、Hcy、hs-CRP水平高于对照组(P<0.05)。不同病情程度脑梗死患者血清ACA、抗β2GP1、Hcy、hs-CRP水平比较:重度组>中度组>轻度组(P<0.05)。相关性分析显示,血清ACA、抗β2GP1、Hcy、hs-CRP水平与老年脑梗死患者病情严重程度均呈正相关关系(r=0.636、0.614、0.563、0.571,P<0.05)。ROC曲线分析显示,血清ACA对轻中度与重度的区分有更高的鉴别价值(P<0.05),敏感度和特异性分别为90.70%、87.10%。结论:老年脑梗死患者血清ACA、抗β2GP1、Hcy、hs-CRP水平均升高,且与病情严重程度正相关,可作为鉴别脑梗死严重程度的有效指标。