Effects and mechanisms of liver cancer cell-derived LC3B + extracellular vesicles on the exhaustion of CD8 + T cells / 中华微生物学和免疫学杂志

Objective:To investigate the potential molecular mechanisms of liver cancer cell-derived secretory autophagosomes, extracellular vesicles expressing LC3B (LC3B + EVs), in promoting the exhaustion of CD8 + T cells. Methods:The proportions of LC3B + EVs and PD-1 + CD8 + T cells in peripheral blood and ascites of liver cancer patients were measured by flow cytometry. Spearman correlation test was used to analyze the correlation between the proportions of LC3B + EVs and PD-1 + CD8 + T cells. Peripheral blood mononuclear cells (PBMCs) from healthy donors were treated with LC3B + EVs or heat shock protein 90α (HSP90α) blocking antibody-pretreated LC3B + EVs for 72 h in the presence of αCD3/CD28 antibodies and IL-2 in vitro. The proportions of PD-1 + CD8 + T and IFN-γ + CD8 + T cells and the concentrations of IL-2, TNF-α and IFN-γ in the supernatants were all detected by flow cytometry. Results:The proportions of LC3B + EVs and HSP90α + LC3B + EVs in plasma and ascites from liver cancer patients were significantly higher than those in healthy control group and non-cancerous ascites group. The level of plasma LC3B + EVs, especially HSP90α + LC3B + EVs, was significantly correlated with the percentage of exhausted PD-1 + CD8 + T cells. In addition, LC3B + EVs from human liver cancer cells up-regulated the percentage of exhausted CD8 + T cells in vitro. However, LC3B + EVs pretreated with HSP90α blocking antibody could significantly inhibit LC3B + EVs-induced CD8 + T cell exhaustion. Conclusions:Liver cancer cell-derived LC3B + EVs could effectively induce CD8 + T cell exhaustion mainly through the membrane-bound HSP90α.

The clinical application of plasmacytoma variant translocation gene 1 in the diagnosis of rheumatoid arthritis / 中华风湿病学杂志

Objective:To explore the clinical application and diagnosis of the long non-coding RNA plasmacytoma variant translocation gene 1 (PVT1) in plasma for rheumatoid arthritis (RA).Methods:One hundred and nineteen healthy individuals were designed as healthy control (HC), 158 patients with RA, 50 patients with systemic lupus erythematosus (SLE) and 50 patients with primary Sj?gren′s syndrome (pSS) were collected from Xuzhou Central Hospital. The plasma PVT1 of HC, RA, SLE and pSS patients and were determined by real time polymerase chain reaction (qRT-PCR). The t test of two independent-samples and One-Way analysis of variance (ANOVA) were used to compare the levels of plasma PVT1 in HC, RA, SLE and pSS patients. The correlation between PVT1 and RF, IL-6 and anti-CCP of RA patients were analyzed by Spearman's rank correlation test. Receiver operating characteristic (ROC) curves were used to identify the diagnostic performance of plasma PVT1 for RA. Results:Compared to HC [(1.32±1.22)], SLE [(1.15±0.83)] and pSS patients [(1.46±0.88)], the plasma PVT1 relative expression [(3.71±2.68)] were significantly increased in RA patients ( t=8.36, P<0.01; t=6.83, P<0.01; t=5.98, P<0.01). The PVT1 had a strong positive correlation with RF, IL-6 and anti-CCP( r=0.41, P<0.01; r=0.38, P<0.01; r=0.40, P<0.01). The area under curve (AUC) of plasma of PVT1 of RA was 0.79[95% CI(0.72, 0.85); P<0.01]. At the optimal cut-off of 1.97, the diagnostic sensitivity and specificity were 68.27% and 86.45%, and in this point provided better diagnostic accuracy. When combination PVT1 with RF, the AUC was 0.88[95% CI(0.83, 0.93); P<0.01], the sensitivity and specificity were 80.22% and 82.73%. Conclusion:Plasma PVT1 has potential diagnostic value for RA, which may become a new biomarker for the diagnosis for RA patients.

Clinical value of long non-coding RNA HOXA terminal transcript antisense RNA in the diagnosis of pancreatic cancer / 中华胰腺病杂志

Objective:To investigate the clinical value of the long non-coding RNA HOXA terminal transcript antisense RNA (HOTTIP) for diagnosing pancreatic cancer (PC).Methods:PC tissue and adjacent normal tissue (>1 cm distant from cancer tissue) from 18 PC patients confirmed by pathology after surgery were collected from June 2017 to December 2018 in Xuzhou Central Hospital. Plasma samples from 78 PC patients clinically confirmed were collected, those from 78 healthy individuals were designed as healthy controls and those from 50 patients of liver cancer, 50 patients of colorectal cancer and 50 patients of gastric cancer were also collected as disease controls. HOTTIP expression in PC tissue and plasma of PC patients, disease controls and healthy controls was tested by real time quantitative polymerase chain reaction; the plasma CA19-9 level was tested by CLIA. The correlation between plasma HOTTIP, cancer tissue HOTTIP and plasma CA19-9 were analyzed, and the relationship between plasma HOTTIP and clinicopathological features was analyzed. The survival curves of patients with high and low expression of HOTTIP were drawn, and the difference of survival rates between the two groups was compared by log-rank test. Receiver operating characteristic (ROC) curves were drawn to calculate area under the ROC curve (AUC), and the diagnostic performance of plasma HOTTIP for PC was evaluated.Results:Compared to normal pancreatic tissue, the level of HOTTIP expression was significantly up-regulated in pancreatic cancer tissue (2.24±0.25 vs 0.62±0.11, P<0.001), the relative expression of plasma HOTTIP of PC, liver cancer, colorectal cancer, gastric cancer patients and healthy controls were 1.33±0.32, 0.57±0.17, 0.51±0.10, 0.41±0.09 and 0.54±0.05; HOTTIP level of PC patients was higher than that of liver cancer, colorectal cancer, gastric cancer patients and healthy controls (all P<0.05), but the difference on HOTTIP level between liver cancer, colorectal cancer, gastric cancer patients and healthy controls was not statistically significant. The plasma HOTTIP of PC patients had a strong positive correlation with plasma CA19-9 and also had a positive correlation with HOTTIP level in cancer tissue (all P<0.05); meanwhile the plasma level of HOTTIP was significantly correlated with TNM stage ( P=0.029), but not with sex, age, lymph node metastasis and tumor size. The median survival time of patients with high HOTTIP level was obviously lower than that of those with low HOTTIP level (15.9 months vs 30.6 months, P<0.05). The AUC of plasma HOTTIP for diagnosing PC was 0.81(95% CI 0.74-0.87). At the optimal cutoff value of 1.14, the diagnostic sensitivity, specificity and accuracy were 62%, 94% and 74%. By combining plasma HOTTIP with CA19-9, the diagnostic sensitivity, specificity and accuracy can be increased to 81%, 97% and 84%, respectively. Conclusions:Plasma HOTTIP level has a significant value in the diagnosis of PC.

The clinical significance of lnc-DC in plasma for the diagnosis of primary Sj(o)gren's syndrome / 中华检验医学杂志

Objective To explore the long non-coding RNA that ineract within dentridic cell (lnc-DC) in plasma as a novel bio-marker for primary Sj(o)grens syndrome (pSS).Methods Case-control study.One hundred and thirty-nine patients of pSS,50 patients of systemic lupus erythematosus(SLE) and 50 patients of rheumatoid arthritis(RA) were collected from Xuzhou Central Hospital from January 2017 to January 2018,and 78 healthy individuals were designed as healthy control.The plasma lnc-DC of pSS,SLE and RA patients and healthy control(HC)was determined by real time polymerase chain reaction (RT-PCR).The plasma levels of anti-SSA(anti-Sj(o)gren's syndrome type A) and anti-SSB(anti-Sj(o)gren's syndrome type B) were tested by ELISA.The correlation between lnc-DC and dry mouth,dry eyes,ESR,β2-microglobulin,anti-SSA and anti-SSB of pSS patients were analyzed by Spearman's rank correlation test.Receiver operating characteristic (ROC) curves were used to identify the diagnostic performance of plasma lnc-DC for pSS.Results Compared to healthy control (3.38 ± 0.44),the lnc-DC relative expression (6.82 ± 0.51) was significantly increased in pSS patients (t=4.78,P＜0.01).The lnc-DC had a strong positive correlation with dry mouth (r=0.81,P＜0.01),dry eyes(r=0.75,P＜0.01),ESR(r=0.73,P＜0.01),β2-microglobulin(r=0.63,P＜0.01),anti-SSA(r=0.45,P＜0.01)and anti-SSB(r=0.53,P＜0.01).The area under curve (AUC) of plasma of lnc-DC was 0.81,(95％ CI:0.71-0.90).When combination lnc-DC with anti-SSA and anti-SSB,the AUC is 0.86 (95％CI:0.78-0.94).At the optimal cut-off of 5.1,the diagnostic sensitivity and specificity were 0.73(95％CI:0.60-0.84) and 0.82 (0.66-0.92).And in this point may provided better diagnostic accuracy.Meanwhile,compared with SLE and RA patients,the plasma level of lnc-DC were higher,ROC curves of lnc-DC in pSS patients can distinguish pSS from other autoimmune disease such as SLE and RA.Conclusions Plasma lnc-DC can provide additional diagnostic information beyond other clinican characteristics such as dry mouth,dry eyes,ESR,β2-microglobulin,anti-SSA and anti-SSB.It can be used as a novel bio-marker for pSS patients,and also have significant values in diagnosis of pSS.

Changes of plasma NGAL and serum CysC in patients with essential hypertension and its clinical significance / 中国基层医药

Objective To explore the levels of neutrophil gelatinase -associated lipocalin(NGAL)and Ser-um cystatin C(CysC)in plasma in essential hypertension(EH)and to discuss their clinical significance.Methods 92 patients with essential hypertension were selected as EH group.At the same time,88 healthy subjects were selected as the healthy control group.The levels of plasma NGAL,serum CysC and some other biochemical markers were detected.The results were statistically processed.Results The level of Plasma NGAL in EH group was(149.22 ± 11.52)μg/L and the level of serum CysC in EH group was(0.92 ±0.03)mmol/L.The level of Plasma NGAL in the healthy control group was(101.4 ±7.71)μg/L and the level of serum CysC in the healthy control group was(0.71 ± 0.02)mmol/L.The levels of Plasma NGAL and serum CysC were significantly increased in EH group compared with those of the healthy control group(F =27.491,P <0.01;F =24.646,P <0.01).Plasma NGAL was positively corre-lated with the degree of EH and the level of serum CysC(r =0.48,P <0.01).Conclusion Plasma NGAL and serum CysC were significantly increased in EH patients,with a significant positive correlation to the degree of nephropathy damage.Both of them can become diagnostic measurements for early diagnosis of nephropathy damage induced by EH.

Relationship between plasma interleukin-18 concentration and cardiovascular risk factors in patients with systemic lupus erythematosus / 国际检验医学杂志

Objective To explore the relationship between interleukin-18(IL-18)and cardiovascular risk factors in patients with systemic lupus erythematosus(SLE).Methods A total of 59 female SLE patients were divided into three groups,according to the IL-18 concentration:≤200 pg/mL(group A),>200-350 pg/mL(group B),>350 pg/mL(group C).The cardiovascular risk fac-tors including body mass index(BMI),systolic blood pressure,diastolic blood pressure(DBP),fasting insulin and glucose,blood lip-id,brachial-ankle pulse wave velocity(baPWV),and plasma homocysteine(Hcy)were determined in all patients.Results Compared with group A patients whose plasma IL-18 level was the lowest,the levels of insulin,triglyceride,Hcy and values of homeostasis model assessment insulin resistance(HOMA-IR)were significantly higher in SLE patients of group C,whose plasma IL-18 level was the highest(P <0.05).Conclusion In patients with SLE,the synergistic effects of hyperinsulinaemia,insulin resistance,hyper-homocysteinaemia,and vascular stiffness most likely contribute to the elevation of plasma IL-18 concentrations.