C-reactive protein to albumin ratio predict responses to programmed cell death-1 inhibitors in hepatocellular carcinoma patients

BACKGROUND Over the years,programmed cell death-1(PD-1)inhibitors have been routinely used for hepatocellular carcinoma(HCC)treatment and yielded improved survival outcomes.Nonetheless,significant heterogeneity surrounds the outcomes of most studies.Therefore,it is critical to search for biomarkers that predict the efficacy of PD-1 inhibitors in patients with HCC.AIM To investigate the role of the C-reactive protein to albumin ratio(CAR)in evaluating the efficacy of PD-1 inhibitors for HCC.METHODS The clinical data of 160 patients with HCC treated with PD-1 inhibitors from January 2018 to November 2022 at the First Affiliated Hospital of Guangxi Medical University were retrospectively analyzed.RESULTS The optimal cut-off value for CAR based on progression-free survival(PFS)was determined to be 1.20 using x-tile software.Cox proportional risk model was used to determine the factors affecting prognosis.Eastern Cooperative Oncology Group performance status[hazard ratio(HR)=1.754,95%confidence interval(95%CI)=1.045-2.944,P=0.033],CAR(HR=2.118,95%CI=1.057-4.243,P=0.034)and tumor number(HR=2.932,95%CI=1.246-6.897,P=0.014)were independent prognostic factors for overall survival.CAR(HR=2.730,95%CI=1.502-4.961,P=0.001),tumor number(HR=1.584,95%CI=1.003-2.500,P=0.048)and neutrophil to lymphocyte ratio(HR=1.120,95%CI=1.022-1.228,P=0.015)were independent prognostic factors for PFS.Two nomograms were constructed based on independent prognostic factors.The C-index index and calibration plots confirmed that the nomogram is a reliable risk prediction tool.The ROC curve and decision curve analysis confirmed that the nomogram has a good predictive effect as well as a net clinical benefit.CONCLUSION Overall,we reveal that the CAR is a potential predictor of short-and long-term prognosis in patients with HCC treated with PD-1 inhibitors.If further verified,CAR-based nomogram may increase the number of markers that predict individualized prognosis.

Over-expression of programmed death-ligand 1 and programmed death-1 on antigen-presenting cells as a predictor of organ dysfunction and mortality during early sepsis: a prospective cohort study

BACKGROUND:This study aimed to explore the changes of programmed death-ligand 1(PDL1)and programmed death-1(PD-1)expression on antigen-presenting cells(APCs)and evaluate their association with organ failure and mortality during early sepsis.METHODS:In total,40 healthy controls and 198 patients with sepsis were included in this study.Peripheral blood was collected within the first 24 h after the diagnosis of sepsis.The expression of PDL1 and PD-1 was determined on APCs,such as B cells,monocytes,and dendritic cells(DCs),by flow cytometry.Cytokines in plasma,such as interferon-γ(IFN-γ),tumor necrosis factor-α(TNF-α),interleukin-4(IL-4),IL-6,IL-10,and IL-17A were determined by Luminex assay.RESULTS:PD-1 expression decreased significantly on B cells,monocytes,myeloid DCs(mDCs),and plasmacytoid DCs(pDCs)as the severity of sepsis increased.PD-1 expression was also markedly decreased in non-survivors compared with survivors.In contrast,PD-L1 expression was markedly higher on mDCs,pDCs,and monocytes in patients with sepsis than in healthy controls and in non-survivors than in survivors.The PD-L1 expression on APCs(monocytes and DCs)was weakly related to organ dysfunction and infl ammation.The area under the receiver operating characteristic curve(AUC)of the PD-1 percentage of monocytes(monocyte PD-1%)+APACHE II model(0.823)and monocyte PD-1%+SOFA model(0.816)had higher prognostic value than other parameters alone.Monocyte PD-1%was an independent risk factor for 28-day mortality.CONCLUSION:The severity of sepsis was correlated with PD-L1 or PD-1 over-expression on APCs.PD-L1 in monocytes and DCs was weakly correlated with infl ammation and organ dysfunction during early sepsis.The combination of SOFA or APACHE II scores with monocyte PD-1%could improve the prediction ability for mortality.

Prognostic value of programmed death.1, programmed death-ligand 1, programmed death-ligand 2 expression, and CD8(+) T cell density in primary tumors and metastatic lymph nodes from patients with stage T1.4N+M0 gastric adenocarcinoma

Background: Anti-programmed death-1/programmed death-ligand 1(PD-1/PD-L1) immunotherapy has been proved to be effective on gastric cancer in ongoing clinical trials. However, the value of PD-L1 in predicting responses of patients with gastric cancer to anti-PD-1/PD-L1 immunotherapy is controversial. Some studies suggested that intra-and inter-tumoral heterogeneity of PD-L1 expression might explain the controversy.This study aimed to analyze the expression of PD-L1, PD-L2, and PD-1 as well as CD8(+) T-cell density in primary tumors and lymph nodes from patients with stage T1-4 N+M0 gastric adenocarcinoma to explore the heterogeneity of PD-1 signaling pathway molecules.Methods: In primary tumors and metastatic as well as non-metastatic lymph nodes from patients with stage T1-4 N+M0 gastric adenocarcinoma, we detected PD-L1 and PD-L2 expression with immunohistochemistry. CD8(+)T-cell density in primary tumors and PD-1 expression on CD8(+)T cells were detected with immunofluorescence. Univariate analysis was used to determine the prognostic values of them. Cox proportional hazard regression model was used to identify independent risk factors that affect patients' overall survival and disease-free survival.Results: Among 119 eligible patients who had undergone surgical resection, the positive rate of PD-L1 was higher in metastatic lymph nodes than in primary tumors(45.4% vs. 38.7%, P = 0.005); the positive rate of PD-1 on CD8(+)T cells was significantly higher in primary tumors and metastatic lymph nodes than in tumor-free lymph nodes(both P < 0.001). The intensity of PD-1 expression on CD8(+) T cells in primary tumors and in metastatic lymph nodes were stronger than that in tumor-free lymph nodes from the same patient. Beside, the positive rate of PD-L2 did not show any differences between primary tumors and metastatic lymph nodes. In multivariate analysis, PD-L1 expression,PD-L2 expression, a low density of CD8(+) T cells in primary tumors, and PD-1 expression on CD8(+) T cells in primary tumors were associated with poor prognosis.Conclusion: The expression of PD-L1 is heterogeneous in primary tumors and in metastatic lymph nodes from patients with stageT1-4 N+M0 gastric adenocarcinoma, which might explain the inconsistent results in assessing the prognostic value of PD-L1 expression in previous studies.

Advances on biological evaluation methods of programmed cell death protein-1/ligand-1 inhibitors

Immuno-oncology represents a groundbreaking and well-established field within cancer treatment.Among the various immuno-oncology targets,the exploration of programmed cell death-1/ligand-1 for drug discovery has proven to be one of the most successful endeavors.Remarkably,it took nearly 30 years from the initial target identification to the clinical approval of monoclonal antibodies.Providing suitable and reliable bioassays for drug candidate evaluation is of paramount importance throughout the early stages of drug discovery,from lead compound identification to in vivo efficacy testing.This assay review aims to shed light on diverse assays reported in the literature for testing antagonism activity and efficacy of programmed cell death-1/ligand-1 inhibitors.Each of these assays possesses inherent advantages and can be applied in different research scenarios.The insights presented in this summary can serve as a valuable resource for scientists in this field,aiding in the selection of appropriate assays for their specific investigations.

Comprehensive insights into the effects and regulatory mechanisms of immune cells expressing programmed death-1/programmed death ligand 1 in solid tumors

The programmed cell death-1(PD-1)/programmed cell death ligand 1(PD-L1)signaling pathway is an important mechanism in tumor immune escape,and expression of PD-L1 on tumor cells has been reported more frequently.However,accumulating evidence suggests that PD-1/PD-L1 is also widely expressed on immune cells,and that regulation is also critical for tumor immune responses.In this review,we emphasized that under solid tumor conditions,the immunoregulatory effects of immune cells expressing PD-1 or PD-L1,affected the prognoses of cancer patients.Therefore,a better understanding of the mechanisms that regulate PD-1 or PD-L1 expression on immune cells would provide clear insights into the increased efficacy of anti-PD antibodies and the development of novel tumor immunotherapy strategies.

The clinical association of programmed cell death protein 4(PDCD4) with solid tumors and its prognostic significance:a meta-analysis

Background: Programmed cell death protein 4(PDCD4) is a novel tumor suppressor protein involved in pro?grammed cell death. Its association with cancer progression has been observed in multiple tumor models, but evidence supporting its association with solid tumors in humans remains controversial. This study aimed to determine the clinical signiicance and prognostic value of PDCD4 in solid tumors.Methods: A systematic literature review was performed to retrieve publications with available clinical informa?tion and survival data. The eligibility of the selected articles was based on the criteria of the Dutch Cochrane Centre proposed by the Meta?analysis Of Observational Studies in Epidemiology group. Pooled odds ratios(ORs), hazard ratios(HRs), and 95% conidence intervals(CIs) for survival analysis were calculated. Publication bias was examined by Begg's and Egger's tests.Results: Clinical data of 2227 cancer patients with solid tumors from 23 studies were evaluated. PDCD4 expression was signiicantly associated with the diferentiation status of head and neck cancer(OR 4.25, 95% CI 1.87–9.66) and digestive system cancer(OR 2.87, 95% CI 1.84–4.48). Down?regulation of PDCD4 was signiicantly associated with short overall survival of patients with head and neck(HR: 3.44, 95% CI 2.38–4.98), breast(HR: 1.86, 95% CI 1.36–2.54), digestive system(HR: 2.12, 95% CI 1.75–2.56), and urinary system cancers(HR: 3.16, 95% CI 1.06–9.41).Conclusions: The current evidence suggests that PDCD4 down?regulation is involved in the progression of several types of solid tumor and is a potential marker for solid tumor prognoses. Its clinical usefulness should be conirmed by large?scale prospective studies.

Programmed cell death-1 inhibitor-related sclerosing cholangitis:A systematic review

BACKGROUND Programmed cell death-1(PD-1)inhibitor has been indicated for many types of malignancies.However,these inhibitors also cause immune-related adverse events.Hepatobiliary disorder is a phenotype of immune-related adverse event affecting 0%–4.5%of patients treated with PD-1 inhibitors.Recent studies have reported PD-1 inhibitor-related sclerosing cholangitis(SC);however,the associated clinical and pathological features are unclear.AIM To evaluate the clinical and pathological features of PD-1 inhibitor-related SC through a systematic review of the literature.METHODS The review,conducted using electronic databases in PubMed,was restricted to the period from January 2014 to September 2019 and focused on case reports/series on PD-1 inhibitor-related SC published in English.We scanned the references of the selected literature to identify any further relevant studies.Six cases previously studied by us,including three that have not yet been published,were included in this review.RESULTS Thirty-one PD-1 inhibitor-related SC cases were evaluated.Median age of patients was 67 years(range,43–89),with a male to female ratio of 21:10.The main disease requiring PD-1 inhibitor treatment was non-small cell lung cancer.Agents that caused PD-1 inhibitor-related SC were nivolumab(19 cases),pembrolizumab(10 cases),avelumab(1 case),and durvalumab(1 case).The median number of cycles until PD-1 inhibitor-related SC onset was 5.5(range,1–27).Abdominal pain or discomfort(35.5%,11/31)was the most frequent symptom.Blood serum tests identified liver dysfunction with a notable increase in biliary tract enzymes relative to hepatic enzymes,and a normal level of serum immunoglobulin G4.Biliary dilation without obstruction(76.9%,20/26),diffuse hypertrophy of the extrahepatic biliary tract(90.5%,19/21),and multiple strictures of the intrahepatic biliary tract(30.4%,7/23)were noted.In 11/23(47.8%)cases,pathological examination indicated that CD8+T cells were the dominant inflammatory cells in the bile duct or peribiliary tract.Although corticosteroids were mainly used for PD inhibitor-related SC treatment,the response rate was 11.5%(3/26).CONCLUSION Some clinical and pathological features of PD-1 inhibitor-related SC were revealed.To establish diagnostic criteria for PD-1 inhibitor-related SC,more cases need to be evaluated.

Immunotherapy against programmed death-1/programmed death ligand 1 in hepatocellular carcinoma: Importance of molecular variations, cellular heterogeneity, and cancer stem cells

Hepatocellular carcinoma(HCC)is a heterogeneous malignancy related to diverse etiological factors.Different oncogenic mechanisms and genetic variations lead to multiple HCC molecular classifications.Recently,an immune-based strategy using immune checkpoint inhibitors(ICIs)was presented in HCC therapy,especially with ICIs against the programmed death-1(PD-1)and its ligand PD-L1.However,despite the success of anti-PD-1/PD-L1 in other cancers,a substantial proportion of HCC patients fail to respond.In this review,we gather current information on biomarkers of anti-PD-1/PD-L1 treatment and the contribution of HCC heterogeneity and hepatic cancer stem cells(CSCs).Genetic variations of PD-1 and PD-L1 are associated with chronic liver disease and progression to cancer.PD-L1 expression in tumoral tissues is differentially expressed in CSCs,particularly in those with a close association with the tumor microenvironment.This information will be beneficial for the selection of patients and the management of the ICIs against PD-1/PD-L1.

Regorafenib combined with programmed cell death-1 inhibitor against refractory colorectal cancer and the platelet-to-lymphocyte ratio’s prediction on effectiveness

BACKGROUND The effectiveness of regorafenib plus programmed cell death-1(PD-1)inhibitor in treating microsatellite stable(MSS)metastatic colorectal cancer(mCRC)remains controversial.AIM To investigate the benefits of regorafenib combined with PD-1 inhibitor in treating MSS mCRC and explore indicators predicting response.METHODS This retrospective study included a total of 30 patients with microsatellite stable metastatic colorectal cancer treated with regorafenib combined with programmed cell death-1 inhibitor at Henan Provincial People’s Hospital between December 2018 and December 2020.During a 4-wk treatment cycle,regorafenib was performed for 3 continuous weeks.PD-1 inhibitor was intravenously injected starting on the first day of the oral intake of regorafenib.We reviewed tumor response,progression-free survival(PFS),overall survival,and treatment-related adverse events(TRAEs)and evaluated association between platelet-tolymphocyte ratio(PLR)and outcomes in this retrospective study.RESULTS Stable disease and progressive disease were found in 18(60.0%)and 12(40.0%)patients,respectively.The disease control rate was 60.0%.The median follow-up time was 12.0 mo,and median PFS was 3.4 mo[95%confidence interval(CI):2.2-4.6 mo].Of the 12 patients with progressive disease,10(83.3%)had liver metastasis before starting the combined treatment.Among the 18 patients with SD,10(55.6%)did not have liver metastases.One patient without liver metastases at baseline was found with a substantially prolonged PFS of 11.2 mo.The liver metastasis,the choice of programmed cell death-1 inhibitor other than nivolumab or pembrolizumab and previous exposure to regorafenib was’t associated with treatment outcome.The median PFS in the low-PLR group was 4.2 mo(95%CI:3.5-4.9 mo),compared with 2.8 mo(95%CI:1.4-4.2 mo)in the high-PLR group(P=0.005).The major TRAEs included hand-foot syndrome(33.3%),hypertension(23.3%),malaise(20.0%),and gastrointestinal reaction(16.7%).The incidence of grade 3 TRAEs was 13.3%(4/30),which comprised abnormal capillary proliferation(n=1),transaminase elevation(n=1),and hand-foot syndrome(n=2).No grade 4 or higher toxicity was observed.CONCLUSION Regorafenib combined with PD-1 inhibitor could lead to a longer PFS in some patients with MSS mCRC.The PLR might be a prediction of the patient response to this therapy.

Programmed cell death protein 4 expression in renal cell carcinoma, penile carcinoma and testicular germ cell cancer

AIM:To investigate the expression of programmed cell death 4(Pdcd4)tumor suppressor gene in tissue specimen of renal cell carcinoma(RCC),testicular germ cell cancer and penile cancer.METHODS:Pdcd4 expression was studied using immunohistochemistry in 188 cases of RCC and 28 controls(including 9 oncocytoma);in 74 cases of penile carcinoma(including 17 metastatic tissue samples)and26 controls;in 11 cases of seminoma,in 14 cases of non-seminoma and 5 controls.RESULTS:Control tissues exhibited strong core and cytoplasmatic Pdcd4 staining.In contrast,core and cy-toplasmatic Pdcd4 levels were significantly decreased in cancer tissues.CONCLUSION:Our data support a role for Pdcd4(down-)regulation in urologic tumors.Interestingly,Pdcd4 expression seem to be a potential diagnostic marker for renal or penile tumors.

PDCD4和miR-107在阴茎癌组织中的表达及与患者预后的关系

目的:探究程序性细胞死亡因子4(PDCD4)及微小RNA-107(miR-107)在阴茎癌组织中的表达水平及与患者预后的关系。方法:选取2018年01月至2020年06月经本院确诊阴茎癌并行手术治疗患者32例,术中分别取患者阴茎癌组织(研究组)及癌旁正常阴茎组织(对照组)各32对,应用实时定量PCR(quantitative reverse transcription-PCR)检测两组PDCD4和miR-107的表达水平并分析其与临床病理参数及预后的关系,并采用Kaplan-Meier进行生存曲线分析及Logistic回归分析影响阴茎癌患者预后的因素。结果:研究组PDCD4表达量明显低于对照组,miR-107表达量明显高于对照组(P均<0.05);PDCD4和miR-107的表达与阴茎癌患者淋巴结转移、组织分级相关(P均<0.05),与年龄、吸烟史、饮酒史、肿瘤部位无关(P均>0.05);应用Kaplan-Meier法对生存率进行分析,miR-107低表达组生存率显著高于miR-107高表达组,PDCD4低表达组生存率低于PDCD4高表达组(P均<0.05)。多因素分析结果显示,TNM分期高、淋巴结转移、PDCD4低表达和miR-107高表达均是影响阴茎癌患者预后的独立危险因素(均P<0.05)。结论:阴茎癌患者miR-107呈异常高表达,PDCD4呈低表达,二者均是影响阴茎癌的独立危险因素且有望作为预测阴茎癌患者预后的重要指标。

下调PDCD4抑制MAP2K3和p38 MAPK的表达减轻LPS诱导的肾小管上皮细胞炎症和凋亡

目的研究程序性细胞死亡蛋白4(programmed cell death protein 4,PDCD4)在脓毒症诱导的急性肾损伤(acute kidney injury,AKI)中的作用机制,以及调控PDCD4表达通过丝裂原活化蛋白激酶3(mitogen-activated protein kinase 3,MAP2K3)和p38蛋白激酶(p38 mitogen-activated protein kinase,p38 MAPK)对脓毒症AKI起到潜在治疗作用。方法用脂多糖(lipopolysaccharide,LPS)刺激人肾小管上皮细胞(HK-2)构建脓毒症AKI细胞模型。进一步用腺病毒介导siRNA和过表达载体抑制和上调AKI细胞模型中PDCD4的表达;CCK-8法检测细胞增殖;用DCFH-DA及激光共聚焦显微镜检测细胞中ROS水平,用总SOD活性检测试剂和MDA检测试剂盒检测细胞中SOD和MDA水平;免疫共沉淀验证PDCD4和MAP2K3之间的蛋白相互作用;TUNEL染色法检测细胞凋亡;RT-qPCR和Western blot检测PDCD4及相关基因的mRNA和蛋白表达水平;ELISA法检测患者血清中炎症相关因子水平。结果LPS诱导可以促进HK-2细胞中PDCD4表达,下调PDCD4可抑制LPS诱导的HK-2细胞的炎症、氧化应激及细胞凋亡。数据库预测及免疫共沉淀证实PDCD4可以与MAP2K3相互作用,且在LPS诱导的HK-2细胞中,MAP2K3表达水平显著增强。MAP2K3过表达和p38 MAPK激动剂可以减轻PDCD4下调对LPS诱导的细胞炎症和氧化应激的影响并抑制细胞凋亡。结论下调PDCD4可以通过抑制MAP2K3和p38 MAPK从而抑制LPS诱导的肾小管上皮细胞的炎症和凋亡。

苍附导痰汤对肥胖型PCOS大鼠内分泌激素及miRNA-16和PDCD-4表达的影响

目的:探讨苍附导痰汤对肥胖型多囊卵巢综合征(PCOS)大鼠内分泌激素及微小RNA(miRNA)-16和程序性细胞死亡因子4(PDCD-4)表达的影响。方法:将50只SPF级雌性SD大鼠随机分为5组,每组10只。阳性对照组大鼠灌胃格华止(0.43 g/kg),低剂量组大鼠灌胃低剂量苍附导痰汤(1.42 g/kg),高剂量组大鼠灌胃高剂量苍附导痰汤(5.68 g/kg),模型组和正常组大鼠灌胃等量生理盐水,各组均持续给药14 d。比较各组大鼠体质量,内分泌激素水平,miRNA-16和PDCD-4 mRNA表达,以及PDCD-4蛋白表达。结果:模型组、阳性对照组、低剂量组和高剂量组大鼠体质量均高于正常组(P<0.05);阳性对照组、低剂量组和高剂量组大鼠体质量均低于模型组(P<0.05),且呈剂量依赖性。模型组、阳性对照组、低剂量组和高剂量组大鼠血清E_(2)均水平低于正常组,而血清T、FSH和LH水平均高于正常组(P<0.05);阳性对照组、低剂量组和高剂量组大鼠血清E_(2)水平均高于模型组,而血清T、FSH和LH水平均低于模型组(P<0.05),且呈剂量依赖性。模型组、阳性对照组、低剂量组和高剂量组大鼠卵巢miRNA-16表达均低于正常组,而PDCD-4 mRNA表达高于正常组(P<0.05);阳性对照组、低剂量组和高剂量组大鼠卵巢miRNA-16表达均高于模型组,而PDCD-4 mRNA表达低于模型组(P<0.05),且呈剂量依赖性。模型组、阳性对照组、低剂量组和高剂量组大鼠卵巢PDCD-4蛋白相对表达量均高于正常组(P<0.05);阳性对照组、低剂量组和高剂量组大鼠卵巢PDCD-4蛋白相对表达量均低于模型组(P<0.05),且呈剂量依赖性。结论:苍附导痰汤可调节肥胖型PCOS大鼠内分泌激素水平,其机制可能与上调miRNA-16表达及下调PDCD-4表达有关。

Combined TIM-3 and PD-1 blockade restrains hepatocellular carcinoma development by facilitating CD4+ and CD8+T cellmediated antitumor immune responses

BACKGROUND Immune checkpoint inhibitors(ICIs)targeting programmed cell death protein 1(PD-1)and T cell immunoglobulin and mucin domain-containing protein 3(TIM-3)are beneficial to the resumption of anti-tumor immunity response and hold extreme potential as efficient therapies for certain malignancies.However,ICIs with a single target exhibit poor overall response rate in hepatocellular carcinoma(HCC)patients due to the complex pathological mechanisms of HCC.AIM To investigate the effects of combined TIM-3 and PD-1 blockade on tumor development in an HCC mouse model,aiming to identify more effective immunotherapies and provide more treatment options for HCC patients.METHODS The levels of PD-1 and TIM-3 on CD4+and CD8+T cells from tumor tissues,ascites,and matched adjacent tissues from HCC patients were determined with flow cytometry.An HCC xenograft mouse model was established and treated with anti-TIM-3 monoclonal antibody(mAb)and/or anti-PD-1 mAb.Tumor growth in each group was measured.Hematoxylin and eosin staining and immunohistochemical staining were used to evaluate T cell infiltration in tumors.The percentage of CD4+and CD8+T cells in tissue samples from mice was tested with flow cytometry.The percentages of PD-1+CD8+,TIM-3+CD8+,and PD-1+TIM-3+CD8+T cells was accessed by flow cytometry.The levels of the cytokines including tumor necrosis factor alpha(TNF-α),interferon-γ(IFN-γ),interleukin(IL)-6,and IL-10 in tumor tissues were gauged with enzyme-linked immunosorbent assay kits.RESULTS We confirmed that PD-1 and TIM-3 expression was substantially upregulated in CD4+and CD8+T cells isolated from tumor tissues and ascites of HCC patients.TIM-3 mAb and PD-1 mAb treatment both reduced tumor volume and weight,while combined blockade had more substantial anti-tumor effects than individual treatment.Then we showed that combined therapy increased T cell infiltration into tumor tissues,and downregulated PD-1 and TIM-3 expression on CD8+T cells in tumor tissues.Moreover,combined treatment facilitated the production of T cell effector cytokines TNF-α and IFN-γ,and reduced the production of immunosuppressive cytokines IL-10 and IL-6 in tumor tissues.Thus,we implicated that combined blockade could ameliorate T cell exhaustion in HCC mouse model.CONCLUSION Combined TIM-3 and PD-1 blockade restrains HCC development by facilitating CD4+ and CD8+T cell-mediated antitumor immune responses.

PDCD4沉默通过调控NLRP3/NLRP6炎症小体的平衡来减轻视网膜缺血再灌注损伤后的神经炎症反应

目的探究程序性细胞死亡因子4(PDCD4)通过含NOD样受体家族Pyrin域蛋白(NLRP)3/NLRP6信号通路活性对视网膜缺血再灌注(RIR)损伤后神经炎症反应的抑制作用及机制。方法选取50只SD大鼠作为研究对象,根据不同处理方式,分为Sham组(空白对照组)、RIR模型组、MCC950组(NLRP3/NLRP6抑制组)、si-PDCD4组(PDCD4沉默组)、si-PDCD4+MCC950组(PDCD4沉默+NLRP3/NLRP6抑制组),每组10只大鼠。除去空白对照组大鼠外,剩余大鼠均构建RIR损伤模型,并进行相应处理;Western-blot检测视网膜组织内PDCD4、NLRP3、NLRP6、天冬氨酸蛋白水解酶-1(Caspase-1)、抗坏血酸过氧化物酶(ASC)表达水平;苏木精-伊红(HE)染色分析视网膜组织病理变化;终端尿苷酸核苷酸末端标记法检测视网膜组织细胞凋亡能力;酶联免疫吸附试验检测大鼠眼球血、视网膜组织内炎症因子白细胞介素(IL)-6、IL-18、IL-1β、肿瘤坏死因子(TNF)-α水平;Western-blot检测视网膜组织内细胞凋亡相关蛋白Bax、Bcl-2、Cleaved-caspase-3表达水平。结果与Sham组比较,RIR模型组PDCD4、NLRP3、NLRP6、Caspase-1、ASC、IL-6、IL-8、IL-1β、TNF-α水平及细胞凋亡指数(AI)均升高,Bcl-2表达水平降低,差异有统计学意义(P<0.05);与RIR模型组比较,MCC950组、si-PDCD4组大鼠NLRP3、NLRP6、Caspase-1、ASC、IL-6、IL-8、IL-1β、TNF-α水平及AI均降低,Bcl-2表达水平升高,差异有统计学意义(P<0.05);与si-PDCD4组及MCC950组比较,si-PDCD4+MCC950组PDCD4、NLRP3、NLRP6、Caspase-1、ASC、IL-6、IL-8、IL-1β、TNF-α水平及AI进一步降低,Bcl-2表达水平进一步升高,差异有统计学意义(P<0.05)。结论PDCD4沉默具有减轻RIR病理损伤,抑制视网膜细胞凋亡的作用,其分子机制可能与抑制NLRP3/NLRP6炎症小体诱导神经炎症反应有关。

妊娠期肝内胆汁淤积症孕妇血清PDCD4,GLUT1表达水平及其与妊娠结局的相关性研究

目的检测妊娠期肝内胆汁淤积症(intrahepatic cholestasis of pregnancy,ICP)孕妇血清程序性细胞死亡因子4(programmed cell death factor 4,PDCD4)和葡萄糖转运蛋白1(glucose transporter 1,GLUT1)的表达水平,分析二者与孕妇妊娠结局的相关性。方法收集巴中市巴州区妇幼保健院产科2018年7月~2020年7月收治的ICP孕妇126例作为研究组,其中轻度ICP组46例,重度ICP组80例,选择同期该院120例健康产检孕妇作为对照组。采用实时荧光定量PCR(qRT-PCR)法测定ICP孕妇血清PDCD4和GLUT1水平,多因素Logistic回归分析影响ICP孕妇妊娠结局的因素,Pearson相关性分析ICP孕妇血清PDCD4和GLUT1水平的相关性。结果与对照组比较,研究组PDCD4(1.36±0.23 vs 1.02±0.21),GLUT1(1.40±0.22 vs 0.99±0.18)水平升高,差异具有统计学意义(t=15.935,12.090,均P=0.000)。重度ICP组PDCD4(1.41±0.25),GLUT1(1.45±0.22)水平显著高于轻度ICP组(1.27±0.20,1.31±0.21),差异具有统计学意义(t=3.246,3.496,均P<0.05)。研究组羊水胎粪污染(20.63%)、自发性早产(7.14%)、产后出血(8.73%)、宫内窘迫(11.90%)等不良妊娠结局的发生率均高于对照组(0.00%,0.83%,0.83%,1.67%),差异均具有统计学意义(χ^(2)=1.049~29.159,均P<0.05);妊娠结局良好组和妊娠结局不良组患者的发病程度(OR=1.109,95%CI=1.035~1.188)、PDCD4(OR=1.428,95%CI=1.013~2.012)以及GLUT1(OR=1.453,95%CI=1.066~1.980)水平差异具有统计学意义(均P<0.05);多因素Logistic回归分析显示,发病程度、PDCD4,GLUT1为ICP孕妇妊娠结局不良的影响因素(Waldχ^(2)==8.738,1.428,1.453;P=0.003,0.041,0.018);Pearson相关性分析显示,ICP孕妇血清PDCD4与GLUT1水平呈正相关(r=0.460,P<0.05)。结论PDCD4,GLUT1在ICP孕妇血清中表达均上调,二者呈正相关,是ICP孕妇妊娠结局不良的影响因素。

lncRNA SNHG1靶向miR-145-5p/PDCD4轴对缺氧/复氧诱导的心肌细胞凋亡的影响

目的探讨长链非编码RNA(lncRNA)核内小RNA宿主基因1(SNHG1)靶向miR-145-5p/程序性细胞死亡因子4(PDCD4)轴对缺氧/复氧(H/R)诱导的心肌细胞凋亡的影响。方法将H9c2细胞分为对照组(NC组)、H/R组、si-NC组、si-SNHG1组、mimic NC组、miR-145-5p mimic组、si-SNHG1+inhibitor NC组、si-SNHG1+miR-145-5p inhibitor组。除NC组外,其他组H9c2细胞均需转染对应物质后构建H/R模型。实时定量PCR(qRT-PCR)检测H9c2细胞中SNHG1、miR-145-5p表达;CCK-8法、流式细胞术分别检测细胞增殖、凋亡;试剂盒检测H9c2细胞中超氧化物歧化酶(SOD)活性、丙二醛(MDA)含量;Western blotting检测H9c2细胞中PDCD4、caspase 3、Bcl-2、Bax蛋白表达;双荧光素酶报告基因实验验证SNHG1与miR-145-5p、miR-145-5p与PDCD4的关系。结果沉默SNHG1或过表达miR-145-5p可促进H/R诱导的H9c2细胞中miR-145-5p表达和细胞增殖,抑制PDCD4蛋白表达、细胞凋亡和氧化应激。miR-145-5p inhibitor减弱了沉默SNHG1对H/R诱导的H9c2细胞中miR-145-5p表达和细胞增殖的促进作用,以及对PDCD4蛋白表达、细胞凋亡、氧化应激的抑制作用。SNHG1靶向调控miR-145-5p/PDCD4轴。结论沉默SNHG1可能通过调控miR-145-5p/PDCD4轴抑制H/R诱导的H9c2细胞凋亡。

miR-155、PDCD4、GSN在高血压性脑出血中的表达及预后价值研究

目的探讨微小核糖核酸(miRNA)-155(miR-155)、程序性细胞死亡蛋白4(PDCD4)、凝溶胶蛋白(GSN)在高血压性脑出血(HICH)中的表达及在预后判断中的价值。方法将2021年1月至2022年1月该院收治的150例HICH患者纳入研究作为研究组。另外,将同期150例无脑出血的高血压患者纳入研究作为对照组。比较研究组、对照组及不同严重程度、不同预后患者miR-155、PDCD4、GSN水平及一般资料。分析各指标与疾病严重程度、预后的相关性。分析HICH预后不良的独立危险因素及各指标单独及联合检测在预后判断中价值。结果研究组miR-155和PDCD4水平均高于对照组,而GSN水平低于对照组(P<0.05)。随HICH严重程度加重,miR-155、PDCD4水平增高,GSN水平降低(P<0.05)。预后不良组miR-155和PDCD4水平均高于预后良好组(P<0.05),而GSN水平低于预后良好组(P<0.05)。miR-155、PDCD4与HICH严重程度呈正相关(r=0.653、0.642,P<0.05),与HICH预后呈负相关(r=-0.625、-0.575,P<0.05);GSN与HICH严重程度呈负相关(r=-0.591,P<0.05),与HICH预后呈正相关(r=0.583,P<0.05)。预后不良组出血量≥60 mL占比高于预后良好组(P<0.05)。miR-155、PDCD4水平升高,GSN水平降低是HICH患者预后不良的危险因素(P<0.05)。miR-155、PDCD4、GSN联合检测用于HICH患者预后不良预测的曲线下面积为0.954,高于miR-155、PDCD4、GSN单独检测(P<0.05)。结论miR-155、PDCD4在HICH患者中呈高表达,GSN呈低表达。miR-155、PDCD4水平升高,GSN水平降低是HICH患者预后不良的危险因素。3项联合检测用于HICH患者预后判断中的价值较高。

血清PDCD4、ZEB1、G-17联合检测对早期胃癌的诊断价值

目的探究血清程序性细胞死亡因子4(PDCD4)、E盒结合锌指蛋白1(ZEB1)、胃泌素-17(G-17)联合检测对早期胃癌(GC)的诊断价值。方法将本院收治的136例GC患者、82例萎缩性胃炎患者分别设为GC组、萎缩性胃炎组,另选取同期136例体检健康者设为对照组。比较各组血清PDCD4 mRNA、ZEB1 mRNA及G-17、糖类抗原19-9(CA19-9)、癌胚抗原(CEA)水平。根据GC患者病情进展程度将其分为早期GC组(80例)和进展期GC组(56例),比较早期GC组、进展期GC组患者血清PDCD4 mRNA、ZEB1 mRNA及G-17、CA19-9、CEA水平;分析血清PDCD4 mRNA、ZEB1 mRNA、G-17、CA19-9、CEA单独或联合检测对早期GC的诊断价值。结果与对照组相比,萎缩性胃炎组、GC组患者血清PDCD4 mRNA表达水平降低(P<0.05),ZEB1 mRNA、G-17水平升高(P<0.05);与萎缩性胃炎组相比,GC组患者血清PDCD4 mRNA表达水平降低(P<0.05),ZEB1 mRNA、G-17、CA19-9、CEA水平升高(P<0.05)。进展期GC组患者血清PDCD4 mRNA表达水平低于早期GC组(P<0.05),ZEB1 mRNA、G-17、CA19-9、CEA水平高于早期GC组(P<0.05)。血清PDCD4、ZEB1、G-17联合诊断早期GC的曲线下面积(AUC)为0.946,高于三者单独诊断及CA19-9、CEA(P<0.05),联合诊断的灵敏度为86.25%,特异度为94.64%,诊断价值最高。结论GC患者血清PDCD4 mRNA表达水平较低,ZEB1 mRNA、G-17水平较高,血清PDCD4、ZEB1、G-17均可辅助诊断早期GC,且三者联合检测可能更有益于临床筛查早期GC。

LncRNA TUG1通过调节miR-181b-5p/PDCD4轴对高糖诱导的心肌细胞凋亡的影响

目的 探讨长链非编码RNA(LncRNA)牛磺酸上调基因1(TUG1)通过调节miR-181b-5p/程序性细胞死亡蛋白4(PDCD4)轴对高糖诱导的心肌细胞凋亡的影响。方法 采用高糖(25 mmol/L葡萄糖)在体外构建糖尿病心肌病(DCM)细胞模型。AC16细胞分为NG组、HG组、HG+sh-NC组、HG+sh-TUG1组、HG+miR-NC组、HG+miR-181b-5p组、HG+sh-TUG1+anti-miR-NC组、HG+sh-TUG1+anti-miR-181b-5p组、HG+miR-181b-5p+pcDNA组、HG+miR-181b-5p+pc-PDCD4组。细胞计数试剂盒-8(CCK-8)法检测细胞活力;乳酸脱氢酶(LDH)测定试剂盒检测LDH释放总量;采用实时定量聚合酶链反应(q RT-PCR)检测TUG1、miR-181b-5p和PDCD4 mRNA表达;流式细胞术检测细胞凋亡;Western blot检测B细胞淋巴瘤2-相关X(Bax)、活化的胱天蛋白酶3(cleaved caspase 3)和PDCD4蛋白表达;caspase-Glo3检测试剂盒评估caspase 3活性;双萤光素酶报告基因实验验证TUG1或PDCD4与miR-181b-5p的靶向关系。结果 与NG组比较,HG组细胞活性降低,LDH释放总量、凋亡率、Bax、cleaved caspase 3表达及caspase 3活性升高(P<0.05),敲低TUG1或上调miR-181b-5p表达可拮抗上述变化(P<0.05)。抑制miR-181b-5p可减轻TUG1沉默对高糖处理下心肌细胞活力和凋亡的影响(P<0.05)。PDCD4过表达可减弱miR-181b-5p上调对高糖处理的心肌细胞活力的促进作用和对凋亡的抑制作用。TUG1可通过吸附miR-181b-5p上调PDCD4表达(P<0.05)。结论TUG1通过下调miR-181b-5p、上调PDCD4表达促进高糖诱导的心肌细胞凋亡。