Better understanding of c-reactive protein and leukocytes in psychiatric inpatients with affective disorders:A biopsychosocial approach

BACKGROUND Affective disorders(AD)have been linked to inflammatory processes,although the underlying mechanisms of this relationship are still not fully elucidated.It is hypothesized that demographic,somatic,lifestyle,and personality variables predict inflammatory parameters in AD.AIM To identify biopsychosocial factors contributing to inflammation in AD measured with two parameters,C-reactive protein(CRP)and leukocytes.METHODS This observational study investigated 186 hospital inpatients diagnosed with AD using demographic parameters,serum inflammatory markers,somatic variables,psychological questionnaires,and lifestyle parameters.Hierarchical regression analyses were used to predict inflammatory markers from demographic,somatic,lifestyle,and personality variables.RESULTS Analyses showed that 33.8%of the variance of CRP was explained by body mass index and other somatic medication(e.g.anti-diabetics),age and education,and age of affective disorder diagnosis.For leukocytes,20.1%of the variance was explained by smoking,diet,metabolic syndrome(MetS),and anti-inflammatory medication(e.g.non-steroidal anti-inflammatory drugs).Other psychiatric or behavioural variables did not reach significance.CONCLUSION Metabolic components seem important,with mounting evidence for a metabolic affective disorder subtype.Lifestyle modifications and psychoeducation should be employed to prevent or treat MetS in AD.

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.

Salivary C-reactive protein and mean platelet volume as possible diagnostic markers for late-onset neonatal pneumonia

BACKGROUND Neonatal sepsis,a formidable threat to newborns,is a leading cause of neonatal mortality,with late-onset sepsis manifesting after 72 hours post-birth being particularly concerning.Pneumonia,a prevalent sepsis presentation,poses a significant risk,especially during the neonatal phase when lung defenses are compromised.Accurate diagnosis of pneumonia is imperative for timely and effective interventions.Saliva,a minimally invasive diagnostic medium,holds great promise for evaluating infections,especially in infants.AIM To investigate the potential of serum C-reactive protein(CRP),salivary CRP(sCRP),and mean platelet volume(MPV)as diagnostic markers for late-onset neonatal pneumonia(LONP).METHODS Eighty full-term neonates were systematically examined,considering anthropometric measurements,clinical manifestations,radiology findings,and essential biomarkers,including serum CRP,sCRP,and MPV.RESULTS The study reveals noteworthy distinctions in serum CRP levels,MPV,and the serum CRP/MPV ratio between neonates with LONP and healthy controls.MPV exhibited a robust discriminatory ability[area under the curve(AUC)=0.87]with high sensitivity and specificity at a cutoff value of>8.8.Correlations between serum CRP,sCRP,and MPV were also identified.Notably,sCRP demonstrated excellent predictive value for serum CRP levels(AUC=0.89),underscoring its potential as a diagnostic tool.CONCLUSION This study underscores the diagnostic promise of salivary and serum biomarkers,specifically MPV and CRP,in identifying and predicting LONP among neonates.These findings advocate for further research to validate their clinical utility in larger neonatal cohorts.

Impact of apolipoprotein E isoforms on sporadic Alzheimer's disease:beyond the role of amyloid beta

The impact of apolipoprotein E(ApoE)isoforms on sporadic Alzheimer's disease has long been studied;however,the influences of apolipoprotein E gene(APOE)on healthy and pathological human brains are not fully understood.ApoE exists as three common isoforms(ApoE2,ApoE3,and ApoE4),which differ in two amino acid residues.Traditionally,ApoE binds cholesterol and phospholipids and ApoE isoforms display diffe rent affinities for their receptors,lipids transport and distribution in the brain and periphery.The role of ApoE in the human depends on ApoE isoforms,brain regions,aging,and neural injury.APOE E4 is the strongest genetic risk factor for sporadic Alzheimer's disease,considering its role in influencing amyloid-beta metabolism.The exact mechanisms by which APOE gene variants may increase or decrease Alzheimer's disease risk are not fully understood,but APOE was also known to affect directly and indirectly tau-mediated neurodegeneration,lipids metabolism,neurovascular unit,and microglial function.Consistent with the biological function of ApoE,ApoE4 isoform significantly alte red signaling pathways associated with cholesterol homeostasis,transport,and myelination.Also,the rare protective APOE variants confirm that ApoE plays an important role in Alzheimer's disease pathogenesis.The objectives of the present mini-review were to describe classical and new roles of various ApoE isoforms in Alzheimer's disease pathophysiology beyond the deposition of amyloid-beta and to establish a functional link between APOE,brain function,and memory,from a molecular to a clinical level.APOE genotype also exerted a heterogeneous effect on clinical Alzheimer's disease phenotype and its outcomes.Not only in learning and memory but also in neuro psychiatric symptoms that occur in a premorbid condition.Cla rifying the relationships between Alzheimer's disease-related pathology with neuropsychiatric symptoms,particularly suicidal ideation in Alzheimer's disease patients,may be useful for elucidating also the underlying pathophysiological process and its prognosis.Also,the effects of anti-amyloid-beta drugs,recently approved for the treatment of Alzheimer's disease,could be influenced by the APOE genotype.

C-reactive Protein/Albumin Ratio as a Prognostic Indicator in Posttraumatic Shock and Outcome of Multiple Trauma Patients

Objective C-reactive protein(CRP)/albumin ratio(CAR)is a new inflammation-based index for predicting the prognosis of various diseases.The CAR determined on admission may help to predict the prognostic value of multiple trauma patients.Methods A total of 264 adult patients with severe multiple trauma were included for the present retrospective study,together with the collection of relevant clinical and laboratory data.CAR,CRP,albumin,shock index and ISS were incorporated into the prognostic model,and the receiver operating characteristic(ROC)curve was drawn.Then,the shock index for patients with different levels of CAR was analyzed.Finally,univariate and multivariate logistic regression analyses were performed to identify the independent risk factors for the 28-day mortality of multiple trauma patients.Results A total of 36 patients had poor survival outcomes,and the mortality rate reached 13.6%.Furthermore,after analyzing the shock index for patients with different levels of CAR,it was revealed that the shock index was significantly higher when CAR was≥4,when compared to CAR<2 and 2≤CAR<4,in multiple trauma patients.The multivariate logistic analysis helped to identify the independent association between the variables CAR(P=0.029)and shock index(P=0.019),and the 28-day mortality of multiple trauma patients.Conclusion CAR is higher in patients with severe multiple trauma.Furthermore,CAR serves as a risk factor for independently predicting the 28-day mortality of multiple trauma patients.The shock index was significantly higher when CAR was≥4 in multiple trauma patients.

Association of C-reactive protein and complement factor H gene polymorphisms with risk of lupus nephritis in Chinese population

BACKGROUND Complement overactivation is a major driver of lupus nephritis(LN).Impaired interactions of C-reactive protein(CRP)with complement factor H(CFH)have been shown as a pathogenic mechanism that contributes to the overactivation of complement in LN.However,genetic variations of neither CRP nor CFH show consistent influences on the risk of LN.AIM To examine whether genetic variations of CRP and CFH in combination can improve the risk stratification in Chinese population.METHODS We genotyped six CRP single nucleotide polymorphisms(SNPs)(rs1205,rs3093062,rs2794521,rs1800947,rs3093077,and rs1130864)and three CFH SNPs(rs482934,rs1061170,and rs1061147)in 270 LN patients and 303 healthy subjects.RESULTS No linkage was found among CRP and CFH SNPs,indicating lack of genetic interactions between the two genes.Moreover,CRP and CFH SNPs,neither individually nor in combination,are associated with the risk or clinical manifestations of LN.Given the unambiguous pathogenic roles of the two genes.CONCLUSION These findings suggest that the biological effects of most genetic variations of CRP and CFH on their expressions or activities are not sufficient to influence the disease course of LN.

Relationship of High Sensitivity C-Reactive Protein with Cardiovascular, Diabetic, and Hepatic Biomarkers

Biomarkers are early predictors of various disorders, circulating level of C-reactive protein is a sensitive biomarker of systemic inflammation and may also be associated with the development of diabetic, hepatic, and cardiovascular diseases. In the present study, we aimed to investigate the association between circulating levels of high sensitive C-reactive protein (hs-CRP) and various biomarkers for hepatic, diabetic, and cardiovascular health. The retrospective analysis included 438 individuals who were tested for these panels simultaneously at Vibrant America Clinical Laboratory. The study population included free-living individuals without any preexisting clinical conditions. Among the cardiovascular markers, a positive correlation and significant association was found between high levels of hs-CRP and serum levels of triglycerides (r = 0.0964, p −0.1423, p −0.1216, p < 0.0105) with circulating levels of hs-CRP. Among all the diabetic markers, glucose (r = 0.1547, p < 0.0011) and glycated serum protein (r = 0.1725, p < 0.0003) were positively correlated with circulating hs-CRP. In the hepatic panel, AST, a transaminase that plays a vital role in amino acid metabolism, was found to have a strong positive correlation with hs-CRP (r = 0.2139, p < 0.0001). In conclusion, the results clearly show the association of hs-CRP with diabetic, hepatic, and cardiovascular risk factors indicating its central value as a key marker for several lifestyle-associated disorders.

Association between Metabolic Syndrome Components and Serum High-Sensitivity C-Reactive Protein or Interleukin-6 Levels among Congolese Adults

Metabolic syndrome (MetS) and its components have been linked to elevated serum levels of inflammatory biomarkers such as C-reactive protein, interleukin-6, interleukin-1β and tumor necrosis factor alpha. The aim of our study was to address the association between MetS components with serum hs-CRP and IL-6 levels among Congolese adults. A total of 357 participants (aged 30 - 87 years) were included in this cross-sectional study. Anthropometrics were collected and fasting blood sampled for assessment of fasting blood glycaemia (FBG), lipids and inflammatory parameters using commercially available assays. NCEP-ATPIII criteria were used to define MetS. The Median (IQR) hs-CRP and IL-6 levels were higher in participants with MetS than in those without ([7 (4, 14) versus 6 (4, 8)] mg/L;p = 0.092 and [23.8 (20.9, 27.6) versus 22.3 (19.5, 25.0)] pg/mL;p = 0.002). hs-CRP and IL-6 levels were significantly higher in females with MetS than in those without, but not in males. Among participants, only TG was correlated with hs-CRP (r = 0.149, p = 0.007), and a significant correlation was observed between TG (r = 0.116, p = 0.037), FBG (r = 0.208, p = 0.000), HDL-C (r = −0.119, p = 0.034) and SBP (r = 0.143, p = 0.010) and IL-6. In males, hs-CRP levels were positively correlated with TG (0.316;p = 0.000), negatively with HDL-C (r = −0.290, p = 0.0022), without such correlations in females. In Ames, IL-6 levels were positively correlated with FBG (r = 0.202;p = 0.035), and negatively with HDL-C (r = −0.249, p = 0.009). Significant correlations between IL-6 levels and FBG (r = 0.214;p = 0.000) or SBP (r = 0.227, p = 0.000) were observed in females. Logistic regression analysis was carried out to identify the relationship between MetS components and hs-CRP or IL-6. Values of area under receiver-operating characteristic (ROC) curves suggest potential use of serum hs-CRP (AUC = 0.675) and IL-6 (AUC = 0.656) as diagnostic biomarkers of MetS. Combination of hs-CRP and IL-6 improved diagnosis accuracy, yielding a 0.698 ROC curve area. MetS components are associated with hs-CRP and IL-6 levels among adults Congolese. Combining the two biomarkers hs-CRP and IL-6 improves Mets diagnostic accuracy compared to hs-CRP or IL-6 alone.

Value of N-Terminal Pro B-Type Natriuretic Peptide,High-Sensitivity C-Reactive Protein,and Homocysteine Levels in Predicting Cardiovascular Events in Chronic Heart Failure Patients After Discharge

Objective:To investigate the value of N-terminal pro B-type natriuretic peptide(NT-proBNP),high-sensitivity C-reactive protein(hs-CRP),and homocysteine(Hcy)levels in predicting cardiovascular events(CV)in patients with chronic heart failure(CHF).Methods:A total of 63 patients with CHF admitted to our hospital between June 2019 and July 2021 were selected.Their NT-proBNP,hs-CRP,and Hcy levels were detected at discharge,and a 12-month follow-up was done after their discharge to collect clinical data.The collected data were inclusive of data from 21 CHF patients with cardiovascular disease and 42 CHF patients without cardiovascular disease.The effect of NT-proBNP,hs-CRP,and Hcy levels on the occurrence of CV was analyzed.Results:The levels of NT-proBNP,hs-CRP,and Hcy in the group with cardiovascular disease were significantly higher than those in the group without cardiovascular disease(P<0.05);the levels of serum NT-proBNP,hs-CRP,and Hcy at discharge had certain value in predicting short-term CV in CHF patients(P<0.05).Conclusion:NT-proBNP,hs-CRP,and Hcy levels can be used to predict CV in CHF patients,thus having clinical application value.

牛病毒性腹泻病毒E^(rns)-ELISA抗体检测试剂盒的制备

为建立快速检测牛病毒性腹泻病毒(BVDV)的血清学方法,本试验以BVDV NADL毒株结构蛋白E^(rns)作为包被抗原,优化反应条件并组装BVDV酶联免疫吸附试验(ELISA)抗体检测试剂盒,通过重复性试验、灵敏性试验、符合率试验和保存期试验验证该试剂盒的准确性、灵敏性和稳定性。结果显示,抗原包被浓度为4μg/mL,1%酪蛋白溶液37℃封闭45 min,被检血清以1∶400稀释孵育30 min,兔抗牛HRP标记二抗以1∶20000稀释孵育30 min,TMB底物反应5 min时,ELISA的反应背景下降,区分度最好。优化后方法的批内重复性试验和批间重复性试验的变异系数均在10%以内,表明该试剂盒具有较好的稳定性;灵敏性试验结果显示,当阳性血清稀释度达到1∶6400时仍可以检测为阳性,说明该试剂盒灵敏度较高;该试剂盒与美国爱德士生物科技公司(IDEXX)BVDV总抗体检测试剂盒共同检测466份临床血清样品,两者总符合率为89.1%;保存期试验结果显示,第4个月时该试剂盒仍能检测到阳性血清,说明稳定性较好。综上表明,本试验利用E^(rns)蛋白建立的BVDV间接ELISA抗体检测试剂盒灵敏度高且重复性好,可为BVDV抗体检测和流行性调查提供新的方法。

Correlation between C-reactive Protein and Morphology of Aortic Intramural Hematoma on CT Angiography

Objectives To investigate the morphologic characteristics of intramural hematoma(IMH)on CT angiography(CTA),and evaluate the possible correlation of serum C-reactive protein(CRP)with morphologic characteristics of IMH.Material and Methods Forty-two patients who were initially diagnosed as IMH by aortic CTA and also had serum CRP examination on the same day of CTA were enrolled in this retrospective study,including 30 males and 12 females,with the mean age of 61±14 years old.The volumetric CT data were retrospectively processed and analyzed on post-processing workstation.Based on the thickness of IMH and the length-area curve,the crosssectional area of true lumen and total vessel were measured,the hematoma-vessel ratio(HVR)was calculated.Imaging characteristics were compared between patients who had pathological elevated CRP(>0.8 mg/dl)and those did not.Spearman correlation analyses of CRP level and morphological characteristics of IMH were performed,and the receiver operating characteristic(ROC)curve was used to evaluate the diagnostic validity of CRP.Results Of all 42 IMH patients,the mean serum CRP was 3.94±4.71 mg/dl,and the mean HVR was 46.7%±14.2%.HVR in patients with elevated CRP was significantly higher than those with normal CRP(49.7%±15.0%vs.40.7%±10.5%,P=0.030).HVR was mildly correlated with CRP in all patients(r=0.48,P<0.001).CRP levels differed neither between patients with Stanford type A and B(P=0.207),nor between patients with and without intimal disruption(P=0.230).To discriminate HVR>47%(the mean value),the area under curve(AUC)were 0.700(95%CI:0.535-0.865)for CRP at a cutoff point of 3.55 mg/dl,with a sensitivity of 54.5%and a specificity of 90.0%.Conclusion CRP was mildly correlated with the severity of cross-sectional hematoma area of IMH,but not with Stanford types and the presence of intimal disruption.

不同程度儿童1型糖尿病酮症酸中毒与血清hs-CRP、IL-10、IgE、NLR水平的相关性分析

目的探讨不同程度儿童1型糖尿病酮症酸中毒血清高敏-C反应蛋白(high-sensitivity C-reactive pro-tein,hs-CRP)、白细胞介素-10(interleukin-10,IL-10)、免疫球蛋白E(serum immunoglobulin E,IgE)、中性粒细胞与淋巴细胞比值(neutrophil to lymphocyte ratio,NLR)水平的相关性。方法回顾性选取2022年1月—2023年5月淄博市妇幼保健院收治的52例儿童1型糖尿病酮症酸中毒患儿的临床资料,进行血清hs-CRP、IL-10、IgE、NLR检测,对检测结果进行相关性分析。结果重度酮症酸中毒患儿血清hs-CRP、IL-10高于轻度、中度患儿,IgE水平低于轻度、中度患儿,差异有统计学意义(P均<0.05)。儿童1型糖尿病酮症酸中毒的严重程度与血清hs-CRP、IL-10呈正相关,与IgE呈负相关(P均<0.05);与NLR无相关性(P>0.05)。结论对于儿童1型糖尿病酮症酸中毒的诊断中,血清hs-CRP、IL-10、IgE均发挥重要作用,NLR对于细菌感染的判断具有积极作用,可更好的对疾病进行诊断,同时监控疾病进展,是诊断和评估病情变化的重要指标。

Prognostic value of C-reactive protein levels within 6 hours after the onset of acute anterior myocardial infarction with primary PCI

Background Increased levels of inflammatory markers have been documented in various settings of coronary artery disease. The vulnerability of coronary lesions in acute myocardial infarction(AMI) at the time of onset may be related to serum levels of C reactive protein(CRP) on admission, before CRP levels are affected by myocardial damage.Objective This study assessed the predictive value of CRP levels within six hours after the onset of acute anterior myocardial infarction with primary percutaneous coronary intervention(PCI).Methods The plasma CRP of 76 patients with first acute anterior myocardial infarction was measured within 6 hours after onset. They were divided into 2 groups: group 1( n =20) with elevated CRP( ≥0.3mg/dl ) on admission within 6 hours after onset and group 2( n =56) with normal CRP( <0.3mg/dl ) within 6 hours after onset. All patients were treated by primary PCI. The primary combined end points, including death due to cardiac causes, re MI related to the infarction artery(RIA) and repeat intervention of the RIA, and the restenosis rate were assessed in relation to CRP levels within 6 hours after onset. Left ventricular end diastolic volume index(EDVI),end systolic volume index(ESVI),and ejection fraction(EF) on admission and 6 month after the onset were assessed by left ventriculography. Changes in EDVI(ΔEDVI),ESVI(ΔESVI), and EF(ΔEF) were obtained by subtracting respective on admission values from corresponding 6 month follow up values. Results There were no significant differences in baseline characteristics between the two groups. The primary combined end points were significantly more frequent in group 1(20%) than those in group 2( 1.79% , P <0.01 ).In addition, restenosis rates were significantly higher in group 1 than in group 2(41.18% vs 16.07%, P<0.05). Group 1 showed greater increases in left ventricular volume and less improvement in EF compared with group 2(ΔEDVI 6.31 ±2.17 vs 3.29 ±9.46ml/m 2 , ΔESVI 5.92 ±2.31 vs 3.86 ±1.08ml/m 2 , ΔEF 1.92 ±0.47 vs 4.79 ±1.73% , P <0.05 , respectively).Conclusions CRP levels within 6 hours after the onset of AMI might predict adverse outcome after primary PCI and progressive ventricular remodeling within 6 month of AMI.

LncRNA ZEB1-AS1和LncRNA SOX2OT在糖尿病肾病患者中的表达及与肾功能的相关性研究

目的探究长链非编码RNA锌指E盒结合同源盒蛋白1反义链1(LncRNA ZEB1-AS1)和长链非编码RNA性别决定相关基因簇2重叠转录本(LncRNA SOX2OT)在糖尿病肾病(DN)患者中的表达及与肾功能的相关性。方法选取于2021年11月—2023年12月在武汉大学人民医院肾内科收治的DN患者106例为DN组,并根据24 h尿蛋白定量(24 h Upro)水平分为正常蛋白尿亚组43例(<30 mg)、微量蛋白尿亚组39例(30~<300 mg)、大量蛋白尿亚组24例(≥300 mg),另选取同期医院单纯糖尿病患者106例作对照组,检测患者血清LncRNA ZEB1-AS1、LncRNA SOX2OT水平;Pearson法分析LncRNA ZEB1-AS1和LncRNA SOX2OT与肾功能指标的相关性;Logistic分析影响DN患者肾功能损伤的因素。结果DN组血清LncRNA ZEB1-AS1、LncRNA SOX2OT水平低于对照组(t=11.471、10.257,P均<0.001)。血清LncRNA ZEB1-AS1、LncRNA SOX2OT比较,正常尿蛋白亚组>微量尿蛋白亚组>大量尿蛋白亚组(F=58.720、117.722,P均<0.001),BUN、SCr、UA水平比较,正常尿蛋白亚组<微量尿蛋白亚组<大量尿蛋白亚组,差异均有统计学意义(F=122.493、595.589、53.178,P均<0.001);LncRNA ZEB1-AS1、LncRNA SOX2OT分别与BUN、SCr、UA呈负相关(r=-0.487、-0.498、-0.521,-0.527、-0.515、-0.534,P均<0.001);Logistic回归分析显示,糖尿病病程长及高BUN、SCr、UA水平是影响DN患者肾功能损伤的危险因素[OR(95%CI)=1.672(1.128~2.479)、2.839(1.534~5.253)、2.754(1.512~5.017)、2.693(1.464~4.954)],高LncRNA ZEB1-AS1、LncRNA SOX2OT是保护因素[OR(95%CI)=0.875(0.798~0.959)、0.898(0.832~0.969)]。结论血清LncRNA ZEB1-AS1、LncRNA SOX2OT水平与DN患者肾功能有关,可能是评估DN患者肾功能的潜在指标。

栀子苷调节PI3K/AKT/mTOR信号通路在动脉粥样硬化形成过程中对Th17/Treg功能的影响

目的:观察栀子苷对载脂蛋白E缺乏(ApoE^(-/-))小鼠Th17/调节性T(Treg)细胞失衡的影响及其作用机制。方法:将50只纯合子ApoE^(-/-)雌性小鼠随机分为对照组、模型组和栀子苷低剂量组、栀子苷中剂量组、栀子苷高剂量组。对照组小鼠喂养普通饲料,模型组和栀子苷组小鼠喂养高脂饲料。从第8周开始,栀子苷各剂量组每日灌胃栀子苷(25、50、100 mg/kg),连续8周。试验结束时,采用油红O染色评估主动脉及其根部动脉粥样硬化(AS)病变面积比。采用定量逆转录聚合酶链式反应(RT-PCR)分析主动脉组织肿瘤坏死因子-α(TNF-α)、白细胞介素(IL)-6、IL-17A和IL-10 mRNA表达;采用流式细胞仪分析脾脏中Th17和Treg细胞百分比;蛋白免疫印迹法(Western Blot)检测主动脉组织磷脂酰肌醇3-激酶(PI3K)/蛋白激酶B(AKT)/哺乳动物雷帕霉素靶蛋白(mTOR)信号通路相关蛋白表达。结果:油红O染色病变显示,栀子苷中剂量组、栀子苷高剂量组病变百分比低于模型组(P<0.05)。与对照组比较,模型组主动脉TNF-α、IL-6和IL-17A mRNA表达水平升高(P<0.05);栀子苷各剂量组主动脉TNF-α、IL-6和IL-17A mRNA表达水平降低(P<0.05)。与对照组比较,模型组主动脉抗炎细胞因子IL-10 mRNA表达水平降低(P<0.05);栀子苷各剂量组主动脉抗炎细胞因子IL-10 mRNA表达水平升高(P<0.05)。与对照组比较,模型组小鼠脾脏中Th17细胞百分比升高,Treg细胞百分比降低(P<0.05)。栀子苷处理恢复了AS小鼠Th17和Treg细胞的平衡。栀子苷抑制PI3K的表达及AKT和mTOR的磷酸化,MHY1485(mTOR活化剂)减弱了栀子苷对T细胞分化的影响。结论:栀子苷抗AS作用机制可能与抑制PI3K/AKT/mTOR信号引起的Treg细胞增多和Th17细胞减少有关。

高糖通过调控miR-429/ZEB1轴对胰腺癌细胞免疫逃逸的影响

目的探究高糖干预对胰腺癌细胞免疫逃逸的影响及作用分子机制。方法采用不同浓度葡萄糖(0、7.5、15、30 mmol/L)处理PANC-1细胞24 h构建高糖干预的PANC-1细胞。将miR-429 mimics及其阴性对照(mimics NC)转染至PANC-1细胞,分为对照组、HG组、HG+mimics NC组、HG+mimics组、HG+mimics+oe-NC组和HG+mimics+oe-ZEB1组。流式细胞术检测细胞表面分子细胞程序性死亡-配体1(PD-L1)表达水平;qRT-PCR检测细胞miR-429和锌指E-盒结合同源盒蛋白1(ZEB1)mRNA表达水平;Western blot检测细胞ZEB1蛋白表达水平。将以上各组PANC-1细胞与CD8^(+)T细胞建立共培养体系,CCK-8检测细胞增殖活性;流式细胞术检测细胞凋亡水平;乳酸脱氢酶(LDH)释放法检测CD8^(+)T细胞对PANC-1细胞的杀伤作用;采用双荧光素酶报告系统验证miR-429和ZEB1的靶向调控关系。结果HG可促进PANC-1细胞表面分子PD-L1及ZEB1表达(P<0.05),抑制miR-429表达,且呈浓度依赖性。miR-429过表达可显著抑制HG诱导的PANC-1细胞表面分子PD-L1表达,而过表达ZEB1可逆转miR-429过表达对HG诱导PANC-1细胞表面分子PD-L1表达的抑制作用。建立与CD8^(+)T细胞共培养体系后,与对照组比较,HG组PANC-1细胞增殖活性明显增加,细胞凋亡率和杀伤活性明显降低(P<0.05);与HG+mimics NC组比较,HG+mimics组PANC-1细胞增殖活性明显降低,细胞凋亡水平和杀伤活性明显升高(P<0.05)。与HG+mimics+oe-NC组比较,HG+mimics+oe-ZEB1组PANC-1细胞增殖活性明显增加,细胞凋亡率和杀伤活性明显降低(P<0.05)。双荧光素酶报告基因实验证实,miR-429靶向负调控ZEB1。结论高糖通过下调miR-429表达水平,靶向负调控ZEB1 mRNA的表达,提高PANC-1细胞表面分子PD-L1表达水平,进而促进PANC-1细胞免疫逃逸。

溶血素 E基因原核表达载体的构建与蛋白诱导表达

目的构建大肠杆菌(Escherichia Coli,E.coli)溶血素E(Hemolysin E)基因的重组质粒pCZN1-hlyE,诱导表达目的蛋白。方法通过NCBI数据库检索获取E.coli Hemolysin E基因序列(NC_000913.3),采用基于PAS(PCR-based Accurate Synthesis)的全基因合成法,设计全长拼接引物,合成目的基因,并成功构建重组质粒pCZN1-hlyE。将构建好的pCZN1-hlyE重组质粒转入大肠杆菌TOP10菌株中,并用异丙基-β-D-硫代半乳糖苷(IPTG)诱导Hemolysin E基因的表达,生成Hemolysin E毒力蛋白。结果经SDS-PAGE凝胶电泳,Western blot分析鉴定目的蛋白Hemolysin E分子量为28.72 kDa,与Hemolysin E基因核酸序列经DNAMAN软件分析所得蛋白分子量一致。结论被诱导表达的Hemolysin E蛋白存在于菌体裂解液上清中,并纯化制备重组蛋白。

miR-150-5p靶向ZEB1调控EMT对子宫内膜癌细胞恶性生物学行为的影响

目的探讨微小RNA-150-5p(miR-150-5p)是否可靶向锌指E盒结合蛋白1(ZEB1)调控子宫内膜癌(EC)细胞上皮间质转化(EMT)进而影响癌细胞的恶性生物学行为。方法RT-qPCR技术检测正常子宫内膜和EC组织中、子宫内膜上皮细胞系hEEC及EC细胞系Ishikawa和HEC-1-A中miR-150-5p相对表达量。过表达miR-150-5p,MTT法、菌落形成实验、伤口愈合实验和Transwell实验分别评估Ishikawa细胞活力、克隆形成、迁移及侵袭能力;双荧光素酶报告基因实验验证miR-150-5p与ZEB1的靶向关系;RT-qPCR检测miR-150-5p及ZEB1 mRNA相对表达量;Western blot技术检测ZEB1、E-钙黏蛋白(E-cadherin)、N-钙黏蛋白(N-cadherin)、波形蛋白(Vimentin)及基质金属蛋白酶9(MMP-9)蛋白表达量。结果与正常子宫内膜组织比较,EC组织中miR-150-5p相对表达量降低(P<0.05);与hEEC细胞比较,HEC-1-A细胞和Ishikawa细胞中miR-150-5p相对表达量降低(P<0.05),Ishikawa细胞中最低(P<0.05)。与空白组比较,miR-150-5p mimics组细胞490 nm处吸光度值、细胞菌落数、迁移数和侵袭数、ZEB1 mRNA和蛋白相对表达量及N-cadherin、Vimentin和MMP-9蛋白相对表达量显著降低,miR-150-5p相对表达量、E-cadherin蛋白相对表达量显著升高(P<0.05)。经生物信息学分析,ZEB1被预测为miR-150-5p的潜在靶基因。结论miR-150-5p可靶向ZEB1抑制癌细胞的恶性生物学行为,其作用机制可能与调控EC细胞EMT进展有关。

Predictive value of C-reactive protein/albumin ratio in acute pancreatitis

BACKGROUND:Serum C-reactive protein(CRP) increases and albumin decreases in patients with inflammation and infection.However,their role in patients with acute pancreatitis is not clear.The present study was to investigate the predictive significance of the CRP/albumin ratio for the prognosis and mortality in acute pancreatitis patients.METHODS:This study was performed retrospectively with 192 acute pancreatitis patients between January 2002 and June 2015.Ranson scores,Atlanta classification and CRP/albumin ratios of the patients were calculated.RESULTS:The CRP/albumin ratio was higher in deceased patients compared to survivors.The CRP/albumin ratio was positively correlated with Ranson score and Atlanta classification in particular and with important prognostic markers such as hospitalization time,CRP and erythrocyte sedimentation rate.In addition to the CRP/albumin ratio,necrotizing pancreatitis type,moderately severe and severe Atlanta classification,and total Ranson score were independent risk factors of mortality.It was found that an increase of 1 unit in the CRP/albumin ratio resulted in an increase of 1.52 times in mortality risk.A prediction value about CRP/albumin ratio >16.28 was found to be a significant marker in predicting mortality with 92.1% sensitivity and 58.0% specificity.It was seen that Ranson and Atlanta classification were higher in patients with CRP/albumin ratio >16.28 compared with those with CRP/albumin ratio ≤16.28.Patients with CRP/albumin ratio >16.28 had a 19.3 times higher chance of death.CONCLUSION:The CRP/albumin ratio is a novel but promising,easy-to-measure,repeatable,non-invasive inflammationbased prognostic score in acute pancreatitis.

Elevated Homocysteine and C-reactive Protein Levels Independently Predict Worsening Prognosis after Stroke in Chinese Patients

Increased plasma total homocysteine (tHcy) and high sensitivity C-reactive protein (hsCRP) levels are independent risk factors for cardiovascular disease.However, the predictive value of tHcy in combination with hsCRP in patients with stroke is not known.To determine the relationship between tHcy and hsCRP, we enrolled 291 patients with first-onset stroke (196 ischemic and 95 hemorrhagic).Plasma tHcy and hsCRP levels were measured and subsequent vascular events and deaths were determined over a 5-year period.Using the arbitrary cutoff for tHcy (【18 μmol/L and ≥18 μmol/L) and hsCRP (【1 mg/L, 1-3 mg/L and 】3 mg/L), the patients were divided into 6 groups.Survival analysis showed that the probability of death or new vascular events during a 5-year follow-up increased according to tHcy and hsCRP levels (P【0.01).The relative risk (RR) of death or new vascular events was 4.67 (95% CI, 1.96 to 11.14, P=0.001) in patients with high tHcy (≥18 μmol/L) and hsCRP (】3 mg/L) compared with those with low tHcy (【18 μmol/L) and hsCRP (【1 mg/L).The increased tHcy level (≥18 μmol/L) combined with increased hsCRP level (】3 mg/L) was still significantly associated with the risk of death or new vascular events (RR, 4.10, 95% CI, 1.61 to 10.45, P=0.003) even when adjusted for other risk factors at inclusion.The combination of increased tHcy and hsCRP levels had a stronger predictive value than increased hsCRP alone or increased tHcy level alone.Further studies are required to evaluate the potential decrease in risks associated with lowering both Hcy and hsCRP levels in patients that present with both increased tHcy and hsCRP.