Objective To investigate the diagnostic efficacy and clinical value of GNB4 and Riplet gene methylation alone and in combination in the diagnosis of primary liver cancer.Methods A total of 313 patients were selected,including 78 patients with primary liver cancer,41 patients with other digestive system tumors,17 patients with non-digestive system tumors,20 patients with postoperative liver cancer,and 157 patients with benign liver disea-ses.The levels of GNB4 and Riplet gene methylation in plasma were detected using quantitative methylation-specific PCR(qMSP).Serum alpha-fetoprotein(AFP)levels were measured by direct chemiluminescence.Results The sensitivity and specificity of AFP in diagnosis were 51.3％and 94.3％,respectively;the sensitivity and specificity of GNB4 gene methylation in diagnosis were 83.3％and 99.4％,respectively;the sensitivity and specificity of Riplet gene methylation in diagnosis were 73.1％and 99.4％,respectively.The sensitivity and specificity of GNB4 and Riplet gene methylation combined diagnosis were 92.3％and 98.7％,respectively;the sensitivity and specificity of AFP,GNB4 and Riplet gene methylation combined diagnosis were 92.3％and 98.7％,respectively;the sensitivity and specificity of combined diagnosis including age and gender were 93.6％and 97.5％,respective-ly.Conclusion The sensitivity and specificity of AFP in the diagnosis of primary liver cancer are limited,while the methylation levels of GNB4 and Riplet genes are higher,and the sensitivity and specificity of their combined de-tection are higher than those of AFP.The sensitivity and specificity of AFP,GNB4 and Riplet gene methylation combined diagnosis are significantly higher than those of AFP,GNB4 and Riplet gene methylation alone.

Objective : To survey the current situation and analyze the relevant influencing factors of sleep quality in adolescent patients with temporomandibular joint anterior disc displacement. Methods: This study has been reviewed and approved by the Ethics Committee, and informed consent has been obtained from patients. A convenience sampling method was used to investigate 120 adolescent patients with temporomandibular joint anterior disc displacement in the outpatient department of stomatology in a grade A tertiary hospital in Shanghai using the general data questionnaire, the Pittsburgh sleep quality index scale (PSQI), the Chinese middle school student mental health scale (MMHI-60) and the pain visual analog scale (VAS). Descriptive analysis, single factor analysis, correlation analysis and multiple regression analysis were used to explore the relevant influencing factors. Results: The PSQI score of adolescent patients with temporomandibular joint anterior disc displacement in this study was 7.77 ± 4.63. There was a statistically significant difference in sleep quality among patients with different academic pressures and levels of sleep bruxism (P<0.05). The sleep quality score was positively correlated with the pain score (r = 0.45, P<0.001) and positively correlated with the psychological score (r = 0.74, P<0.001). The degree of pain can affect the patient's sleep quality, and those with good mental health have better sleep quality. The results of regression analysis showed that academic stress (OR = 2.511, 95% CI =1.307 ~ 4.828), bruxism (OR = 3.694, 95% CI = 1.394 ~ 9.791), pain score (OR = 2.104, 95% CI =1.095 ~ 4.041) and psychological score (OR = 1.039, 95% CI = 1.021 ~ 1.058) were statistically significant. Conclusion The sleep quality of adolescent patients with temporomandibular joint anterior disc displacement is generally poor. Academic pressure, sleep bruxism, pain and mental health are the influencing factors of sleep quality.

Objective:To investigate the expression and clinical significance of neutrophil myeloperoxidase (MPO) in patients with MPO-antibody associated vasculitis (AAV).Methods:Thirty-six newly diagnosed MPO-AAV patients who were hospitalized in the First Affiliated Hospital, Anhui Medical University from July 2018 to June 2021 were enrolled,and 36 age and sex matched healthy subjects were selected as controls. Neutrophil MPO level was detected by flow cytometry (FCM) and MPO mRNA was tested by real time quantitative polymerase chain reaction (RT-qPCR) in all subjects. Serum complement fragment C5 (C5a) and MPO in both groups and serum MPO-anti-antineutrophilic cytoplasmic antibody(ANCA) in MPO-AAV group were measured by enzyme linked immunosorbent assay (ELISA), while the disease activity was evaluated by Birmingham vasculitis activity score-V3 (BVAS-V3).Results:Compared with the heathy control group, the expression of MPO mRNA in neutrophils, serum MPO and complement C5a in MPO-AAV group were significantly higher[MPO mRNA:30.2±11.5 vs. 1.9±0.6, P<0.001;MPO:(112.0±68.7) IU/L vs. (87.4±22.9) IU/L, P=0.01; C5a:(187.3±90.3) ng/ml vs. (107.3±31.1) ng/ml, P<0.001; respectively], while the mean fluorescence intensity (MFI) of MPO in neutrophils were significantly lower [ 1 343.3±723.4 vs. 2 868.0±1 136.5, P<0.001]. In MPO-AAV group, the expression of neutrophil MPO mRNA was positively correlated with serum MPO-ANCA and MPO levels ( r=0.537, P=0.001 and r=0.358, P=0.032; respectively). Multiple regression analysis suggested that neutrophil MPO mRNA expression was positively correlated with serum MPO-ANCA level ( β=0.695, P=0.006); neutrophil MPO level was negatively correlated with serum MPO-ANCA, MPO and complement C5a levels ( r=-0.335, P=0.046; r=-0.372, P=0.026; r=-0.577, P<0.001; respectively). Further, neutrophil MPO level was negatively correlated with serum complement C5a level ( β=-0.374, P=0.043). BVAS-V3 was positively correlated with MPO mRNA expression in neutrophils, serum MPO-ANCA, MPO and complement C5a ( r=0.598, P<0.001; r=0.599, P<0.001; r=0.537, P=0.001; r=0.415, P=0.012; respectively) and negatively correlated with MPO level in neutrophils ( r=-0.342, P=0.041). In multiple regression analysis it suggested that BVAS-V3 was positively correlated with MPO mRNA expression in neutrophils ( β=0.511, P=0.002). Conclusion:In MPO-AAV patients, MPO synthesis and release in neutrophils are both significantly increased, which might be influenced by serum MPO-ANCA and C5a, respectively. Furthermore, MPO synthesis activity in neutrophils is an independent factor related to disease activity.

Objective:To explore the related indexes of coagulation and thrombosis and their clinical significance in patients with severe fever with thrombocytopenia symptoms (SFTS) during the onset and recovery period after novel bunyavirus infection.Methods:A total of 36 patients diagnosed with SFTS (SFTS onset group) and 18 convalescent SFTS patients, who were hospitalized in the First Affiliated Hospital of Anhui Medical University from April 12, 2020, to October 12, 2020 were recruited in this study. Thirty-six healthy controls were recruited from volunteers. Plasma was collected and prothrombin time (PT), activated partial thromboplastin time (APTT), fibrinogen (FIB), thrombin time(TT), antithrombin-Ⅲ (AT-Ⅲ), fibrin degradation product (FDP) and D-dimer (D-D) were determined by automatic blood coagulation analyzer. Thrombomodulin (TM), thrombin-antithrombin complex (TAT), plasminase-α2 plasminase inhibitor complex (PIC) and tissue plasminogen activator-plasminogen activator inhibitor 1 complex (t-PAIC) were determined by an automatic chemiluminescence analyzer.Results:Compared with the healthy control group, PT was significantly prolonged (12.5 [12.1, 13.6] s, vs 10.8 [10.5, 11.5] s, P<0.05) in SFTS onset group, but was still within the reference range (14.0-21.0 s), and APTT (49.1 [42.0, 58.2]s vs 28.5 [26.6, 30.4]s, P<0.05) was also significantly prolonged in SFTS onset group. Compared with healthy control group, FDP (6.07 [2.67, 8.64] μg/ml vs 1.00 [0.80, 1.87] μg/ml, P<0.001), D-D (2.27 [1.04, 2.98] μg/ml vs 0.30 [0.21, 0.47] μg/ml, P<0.001), TAT (16.05 [8.05, 26.58] ng/ml vs 3.55 [2.60, 4.85] ng/ml, P<0.001), PIC (4.44 [2.52, 5.54] μg/ml vs 0.84 [0.60, 1.35] μg/ml, P<0.001), TM ([19.41±8.29] TU/ml vs [9.33±1.89] TU/ml, P<0.001), and t-PAIC ([37.52±21.10] ng/ml vs [7.06±3.37] ng/ml, P<0.001) values were all significantly higher in the SFTS onset group (all P<0.001). The level of TAT in the SFTS recovery group (9.10 [3.95, 18.40] ng/ml) was still out of the reference range (<4 ng/ml), while the level of PIC in the SFTS recovery group was lower than in SFTS onset group (1.91 [1.45, 2.93] μg/ml vs 4.44 [2.52, 5.54] μg/ml, P<0.05). Compared with SFTS onset group, the levels of TM and t-PAIC were lower in the SFTS recovery group ( P<0.05). Conclusions:Coagulation system activation and vascular endothelial injury are evidenced in SFTS patients. In the convalescence period, the vascular endothelial injury is recovered, however, there is still a certain degree of coagulation dysfunction, therefore, it is necessary to monitor the coagulation indicator of discharged patients post SFTS.

Objective: To explore the high expression level of serum heat shock protein 90α(HSP90α) and gene HSP90 AA1 in cancer tissue, and to discover the prognosis of lung cancer. Methods: A total of 109 cases of lung cancer were collected as the experimental group; 38 lung inflammation groups as the reference group; and 30 healthy controls. The serum HSP90α levels between the three groups were compared; the correlation between HSP90α and clinical parameters were analyzed. The TCGA data were used to analyze the correlation between the expression level of HSP90 AA1 and various pathological features, as well as its influence on the prognosis of lung cancers. Results: The serum HSP90α concentrations in the experimental group were higher than those of the reference group and the control group(P<0.05). The ROC curve area of HSP90α in the diagnosis of lung cancer was 0.898(P<0.05); the expression of HSP90α in the lung squamous cell carcinoma group(LUSC) and the small cell lung cancer group(SCLC) were higher than those in lung adenocarcinoma group(LUAD)(P<0.05); the level of HSP90α decreased when condition alleviated, while increased significantly when disease progressed(P<0.05); Further TCGA database showed that the expression of HSP90 AA1 in cancer tissues was higher than that of adjacent cancer tissues(P<0.05),meanwhile, remarkably increased in LUSC compared with in LUAD(P<0.05). The expression of HSP90 AA1 has no significant correlation with the age, gender, clinical stage, and tumor residue of lung cancer patients. Survival analysis and Cox regression analysis showed that high expression of HSP90 AA1 reduced the overall survival(OS) of lung cancer patients; HSP90 AA1 was an independent prognostic factor for lung cancer patients. Conclusion Serum HSP90α and HSPAA1 in cancer tissues are elevated in patients with lung cancer, which should be used as an auxiliary diagnosis method for lung cancer. Meanwhile they should be used for therapeutic effect observation and survival status prediction.

Objective:To investigate the change of peptidylarginine deiminase 4 (PAD4) expression in the neutrophils of peripheral blood of patients with anti-neutrophil cytoplasmic myeloperoxidase antibody-associated vasculitis (MPO-AAV), and to analyze the relationship between the change and disease activity.Methods:Thirty-nine untreated patients with active MPO-AAV (patient group) and thirty-nine healthy volunteers (control group) were enrolled into this study. The PAD4 expressed on neutrophils was detected by flow cytometry (FCM). The serum neutrophil extracellular traps (NETs), fragments from the activated complement C5 (C5a) and anti-neutrophil cytoplasmic myeloperoxidase antibody (MPO-ANCA) were measured by enzyme-linked immuno sorbent assay (ELISA). Their disease activity was evaluated by Birmingham vasculitis activity score (BVAS). All the detected results were compared between the 2 groups by t test, Spearman correlation analysis and multivariate linear regression analysis were employed to analyze the relationship between BVAS and the Lab test results in patient group. Results:The proportion of neutrophil expressing PAD4, the mean fluorescence intensity (MIF) of PAD4, the levels of NETs and C5a in patient group were significantly higher than those in the control group [(71±11)% vs (26±6)%, t=22.456, P<0.01; (33±4) vs (14±4), t=18.668, P<0.01; (0.62±0.22) vs (0.26±0.15), t=8.466, P<0.01 and (4.6±1.0) vs (2.9±1.0), t=7.697, P<0.01, respectively]. In patient group, BVAS was positively associated with the proportion of PAD4 + neutrophil, MFI of PAD4, the serum level of NETs, C5a and MPO-ANCA ( r=0.843, P<0.01; r=0.821, P<0.01; r=0.411 1, P<0.01; r=0.613, P<0.01 and P=0.790, P<0.01, respectively), however, the results of multiple linear regression analysis showed only the percentage of PAD4 + neutrophils and the level of MPO-ANCA were independent influencing factors on BVAS ( β=0.324 6, P<0.01 and β=0.796, P<0.01, respectively). Conclusion:The percentage of neutrophils expressed PAD4 in the peripheral blood of patient with active MPO-AAV is significantly increased, and it is an independent factor affecting the disease activity. Intervention on this expression might be a potential new pathway for MPO-AAV treatment.

BACKGROUND: The incidence of lung cancer is increasing annually. Clinicians pay special attention to lung tests during physical examinations. Due to the popularity of low-dose computed tomography, not only can lung cancer be diagnosed early, but physical examinations often reveal the presence of pulmonary nodules, an important health issue that cannot be ignored. Patients with pulmonary nodules are prone to adverse emotions such as anxiety and depression. Many studies have shown that patients with emotional disorders have immune system dysfunction and changes in inflammation levels. This study aimed to investigate the changes in anxiety, depression, the ratios of T helper cells 17 (Th17) and regulatory T cells (Tregs), and inflammation levels in patients with pulmonary nodules. METHODS: A total of 143 subjects from The First Affiliated Hospital of Anhui Medical University were included from April 2019 to July 2019. All of the subjects were assessed with the Beck Anxiety Inventory (BAI) and the Beck Depression Inventory-II (BDI-II). Overall, 40 cases were healthy controls (HC) and 103 cases were patients with pulmonary nodules. The patients were divided into two groups according to the scale scores: 62 cases in a non-anxiety and non-depression (NAD) group and 41 cases in an anxiety and/or depression (AD) group. The percentage of Th17 and Tregs in the peripheral blood and inflammatory factors in the serum were detected. The absolute Th17 cell counts were calculated and the differences between the groups and correlations between these indicators were analyzed. RESULTS: There were statistically significant differences in the percentage of Th17 cells, the absolute counts of Th17 and Th17/Treg cells, and the levels of interleukin-2 (IL-2), IL-6, and tumor necrosis factor-α (TNF-α) among three groups (all P<0.001). The AD group was higher than the HC and NAD groups (all P<0.05). There was no statistically significant difference between the HC and NAD groups (all P>0.05). The previously described indicators had no significant correlation with the severity of anxiety and depression (P>0.05). There were no significant differences in the percentage of Tregs or levels of IL-4 and IL-10 between the groups (all P>0.05). The proportion of anxiety and/or depression in female patients with pulmonary nodules was higher than that in males (P<0.05). CONCLUSIONS Patients with pulmonary nodules are prone to varying degrees of anxiety and depression, which leads to immune dysfunction and low-grade inflammation.

Objective To produce a plastic ampoule with an opening that fits syringe adapters to allow solutions to be drawn up without needles (to avoid needle stick and sharps injuries) and to test the effectiveness of the new design in terms of residual fluid volume and risk of microorganism contamination. Methods Evaluation study based on laboratory research was adopted. For the experimental group, 30 ampoules based on the new design were produced by 3D printing. The ampoules were sterilized and filled with 2 ml sterile water. The sterile water was drawn up without the use of needles, and the wastage fluid (i.e., 2 ml minus the quantity drawn up) was calculated. A 1 ml aliquot of sterile water from each ampoule was dropped onto a nutrient agar plate, and the number of colony-forming units was assessed after 48 h. For each ampoule, the experiment was performed twice. Sixty 2 ml glass-packaged sterile water injections constituted the control group. The fluid was drawn up with a needle, and the superfluous fluid and number of colony-forming units were assessed, as in the experimental group. Results The mean wastage fluid was 0.06 ml, 95% CI was 0.05-0.07 ml in the experimental group and the superfluous fluid was 0.06 ml, 95% CI was 0.06-0.07 ml in the control group, there was no significant difference between the two groups (Z=-1.194, P=0.233). The number of colony-forming units was 8 in the experimental group and 4 in the control group, there was no significant difference between the two groups (P=0.224). Conclusions The newly designed plastic ampoule opening could help health workers to avoid needle stick and sharps injuries when drawing up solution. The wastage fluid and microorganism contamination levels met the required standards, indicating that the new design is suitable for clinical application.

Objective To investigate the change of circulating follicular helper T cells (cTfh) in patients with anti-neutrophil cytoplasmic myeloperoxidase antibody-associated vasculitis (MPO-AAV), and to analyze the relationship between cTfh and disease activity. Methods Thirty-eight untreated MPO-AAV patients (patient group) and thirty-eight healthy volunteers (control group) were enrolled in this study. cTfh and membrane expression of inducible co-stimulator(ICOS)and programmed cell death protein 1(PD-1) were detected by flow cytometry (FCM). Serum anti-neutrophil cytoplasmic myeloperoxidase antibody (MPO-ANCA) was measured by ELISA. Disease activity was evaluated by Birmingham vasculitis activity score (BVAS). Results Compared with those in control group, the proportions of cTfh, ICOS+Tfh and PD-1+Tfh cells in patient group were significantly higher [(25.9±3.8)％vs. (21.0±5.3)％, P<0.001;(1.8±0.8)％vs. (0.8±0.5)％, P<0.001 and (10.2±2.8)％vs. (8.2±2.2)％, P=0.001, respectively]. Meanwhile, the expression of ICOS and PD-1 on cTfh in patient group was markedly more intensive (59.6±10.0 vs.49.2±6.9, P<0.001 and 532.6±104.2 vs. 485.1±73.4, P=0.025, respectively). In patient group, the proportion of cTfh was positively correlated with the ratio of ICOS+Tfh, the expression of ICOS, the level of MPO-ANCA and BVAS (r=0.407, P=0.011; r=0.705, P<0.001; r=0.737, P<0.001 and r=0.663, P<0.001, respectively). The expression intensity of ICOS on cTfh was positively associated with ICOS+Tfh ratio, serum MPO-ANCA and BVAS (r=0.388, P=0.016; r=0.645, P<0.001 and r=0.653, P<0.001, respectively). Nevertheless, the expression of PD-1 on cTfh was only positively correlated with the ratio of PD-1+Tfh (r=0.473, P=0.003). Conclusions Enhanced cTfh in patients with MPO-AAV might produce MPO-ANCA, which is related to the aggravation of MPO-AAV. Thus, cTfh and its ICOS could be potentially targeted for the treatment of MPO-AAV.

Objective To investigate the change of CD35 expression on neutrophils in the peripheral blood and the relationship between the change and disease activity in patient with myeloperoxidase antineutrophil cytoplasmic antibody-associated vasculitis (MPO-AAV).Methods Forty untreated patients with active MPO-AAV(patient group)and forty healthy volunteers (control group) were enrolled into this study,and Bermingham vasculitis activity score (BVAS) for every patient was recorded.Flow cytometry (FCM) was employed to detect the CD35 and MPO expression on the neurtrophil,and enzyme linked immunosorbent assay (ELISA) was taken to test the levels of autoantibody against MPO-Antineutrophil cytoplasmic antibody (MPO-ANCA),fragment a from the activated complement factor B (Ba) and MPO in peripheral blood from both group.All test results were compared between the 2 groups by t test,Non-parametric test,Spearman correlation analysis.In addition,the relations among the laboratory results and the relationship between BVAS and the laboratory results were analyzed respectively.Results Compared with the control group,the expression level,which was represented as mean flourscence indensity (MFI),of CD35 and neutrophil membrane MPO on peripheral blood neutrophils was significantly increased [(2 014±968) vs (1 454±511),t=3.024,P=0.002 and (709±244) vs (580±158),t=2.806,P＜0.01,respectively],and the MPO expression level in neutrophils was significantly lower [(1 525±1 033) vs (3 196±2 126),t=-4.468,P＜0.01].Ba and MPO levels in serum of the patient group was significantly higher than that in the control group [37.89(26.17,63.14) μg/L vs 27.99(18.64,46.52) μg/L,Z=-2.521,P=0.012 and 546.16(450.55,729.96) U/L vs 327.93(279.02,365.10) U/L,Z=7.121,P＜0.01,respectively].In patient group,the expression level of CD35 had a significant positive relationship with peripheral blood neutrophil count (r=0.573,P＜0.01),serum Ba (r=0.433,P=0.005) and BVAS (r=0.368,P=0.020),respectively,whereas,there was a negative correlation between the MPO expressed on the neutrophils and that in the neutrophils (r=-0.458,P=0.003),and a positive relationship between MPO-ANCA and BVAS (r=0.351,P=0.026).Conclusion There is significant increased expression of CD35 on the neutrophil of patient with MPO-AAV,which might protect the neutrophil from destruction by the activated complement alternative pathway,and more neutrophils consequently contribute to the MPO-AAV pathogenesis.Inhibition of CD35 expression might become one of the potential new pathways for the treatment of MPO-AAV.