Abnormal coagulation activity and the related clinical significance in patients with novel bunyavirus infection / 中华检验医学杂志

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.

The expression and clinical significance of neutrophil myeloperoxidase in patients with myeloperoxidase-antineutrophil cytoplasmic antibody associated vasculitis / 中华内科杂志

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.

A preliminary study on the change and clinical significance of peptidylarginine deiminase 4 expression on the neutrophils in the peripheral blood from the patients with anti-neutrophil cytoplasmic myeloperoxidase antibody-associated vasculitis / 中华风湿病学杂志

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.

Correlation between Anxiety, Depression and Changes in Th17/Treg and Inflammatory Levels in Patients with Pulmonary Nodules / 中国肺癌杂志

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 P0.05). 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.

The clinical significance of circulating follicular helper T cells in patients with anti-neutrophil cytoplasmic myeloperoxidase antibody-associated vasculitis / 中华内科杂志

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.

A preliminary study on the change and significance of CD35 expression on the neutrophils of per-ipheral blood from patient with myeloperoxidase-antineutrophil cytoplasmic antibody associated vas culitis / 中华风湿病学杂志

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.

Natural killer cells in liver diseases / 医学前沿

The liver has been characterized as a frontline lymphoid organ with complex immunological features such as liver immunity and liver tolerance. Liver tolerance plays an important role in liver diseases including acute inflammation, chronic infection, autoimmune disease, and tumors. The liver contains a large proportion of natural killer (NK) cells, which exhibit heterogeneity in phenotypic and functional characteristics. NK cell activation, well known for its role in the immune surveillance against tumor and pathogen-infected cells, depends on the balance between numerous activating and inhibitory signals. In addition to the innate direct "killer" functions, NK cell activity contributes to regulate innate and adaptive immunity (helper or regulator). Under the setting of liver diseases, NK cells are of great importance for stimulating or inhibiting immune responses, leading to either immune activation or immune tolerance. Here, we focus on the relationship between NK cell biology, such as their phenotypic features and functional diversity, and liver diseases.

Effect of miRNA-143 on proliferation, migration and invasion of CNE-2Z cells of human nasopharyngeal cancer / 安徽医科大学学报

Objective To detect the effects of miR-143 on proliferation, migration and invasion of human nasopharyngeal cancer CNE-2Z cells.Methods The expression of miR-143 in NP69 cells, CNE-1 cells and CNE-2Z cells were detected by using real-time PCR.The overexpression of miR-143 in CNE-2Z cells was constructed through infecting lentivrius, and the level of miR-143 was determined by using real-time PCR.Cell viability was detected by using a CCK-8 kit according to the manufacturer's instruction at indicated time points.The effectiveness of overex-pression of miR-143 on migration and invasion of CNE-2Z cells were detected by using Transwell cell migration and invasion assay.Results The expression level of miR-143 in CNE-2Z was significantly lower than those in NP69 and CNE-1 (P<0.01).The miR-143 over-expressed CNE-2Z cell was successfully established.The cell viability in CNE-2Z/ miR-143 group was significantly decreased compared with CNE-2Z and CNE-2Z/miR-NC (P<0.01).Overexpression of miR-143 inhibited cell migration and invasion of CNE-2Z cells significantly(P<0.01).Conclusion miR-143 might inhibit cell proliferation, migration and invasion in human nasopharyngeal cancer CNE-2Z cells, indicating its important role in diagnosis, treatment and prognosis of nasopharyngeal cancer.

The significances of peripheral neutrophils CD55 and myeloperoxidase expression in patients with myeloperoxidase-specific anti-neutrophil cytoplasmic antibody associated vasculitis / 中华内科杂志

Objective To investigate the expression of CD55 and myeloperoxidase (MPO) on neutrophils in patients with MPO-specific anti-neutrophil cytoplasmic antibody associated vasculitis(MPO-AAV), and analyze the relationship between the expression and clinical manifestation.Methods Forty untreated patients with active MPO-AAV (patient group) and 30 healthy volunteers (control group) were enrolled in this study.The CD55 on neutrophils and both membrane and cytoplasmic MPO were detected by flow cytometry.Serum fragment-from the activated complement factor B(Ba) and MPO were measured by ELISA.The clinical activity of vasculitis was valued by Birmingham vasculitis activity score-version 3(BVAS-V3).The significance of laboratory data was evaluated by Spearman correlation test and multivariate linear regression analysis.Results (1)The mean fluorescence intensity(MFI) of CD55 expressed on neutrophils was significantly higher than that in control group[4 068.6±2 306.0 vs 2 999.5±1 504.9,P=0.033].Similar results of serum MPO and Ba in patient group were found compared to controls [500.0(381.0, 612.7) IU/L vs 286.9(225.5, 329.1) IU/L,P<0.001;35.2(25.2, 79.5) ng/L vs 18.0(15.0, 28.0) ng/L,P<0.001], respectively.However, MIF of cytoplasmic MPO in patients was significantly lower than that of control group(1 577.1±1 175.9 vs 3 105.3±2 323.0,P=0.003).(2) In patient group, cytoplasmic intensity of MPO was negatively associated with the serum levels of MPO(r=-0.710,P<0.001) and Ba (r=-0.589,P=0.001).Moreover, serum MPO was positively associated with serum Ba(r=0.691,P<0.001).Membrane intensity of CD55 on neutrophils was positively correlated with patient age (r=0.514, P=0.001), C reactive protein (r=0.376, P=0.018), peripheral neutrophils count (r=0.485, P=0.001) and BVAS-V3 (r=0.484, P=0.002), whereas negative correlation between membrane CD55 and disease duration was seen (r=-0.403,P=0.01).(3) The result of multiple linear regression analysis showed there was statistically significant positive correlation between MFI of CD55 expressed on neutrophils and BVAS-V3 (β=0.001,P=0.027).Conclusions In MPO-AAV,CD55 expression on neutrophils is markedly enhanced, which is one of the independent risk factors related to disease activity.It might protect neutrophils from attacking AAV, CD55 expression on neutrophils is markedly enhanced, which is one of the independent risk factors related to disease activity.It might protect neutrophils from attacking by complement alternative pathway.Activated neutrophils release more MPO and lysosome to intensify the inflammation reaction and aggravate the disease.Thus CD55 might become a new potential target for the treatment of this disease in the future.

Comparative analysis of hepatitis B virus large protein, hepatitis B virus-DNA and Pre-S1 antigen in evaluating serum hepatitis B virus replication / 中华预防医学杂志

Objective@#To investigate the comparative study of serum hepatitis B virus (HBV) large protein (HBV-LP) , HBV-DNA, and Pre S1 antigen (Pre S1-Ag) detection in the evaluation of serum HBV replication in patients with chronic hepatitis B.@*Methods@#A total of 482 patients infected with chronic hepatitis B virus (CHB) were enrolled and the serums were collected in a hospital of Hefei city in Anhui province from June 2013 to March 2015. The serum HBV-LP, HBV markers(HBV-M) and Pre S1-Ag were detected using ELISA, and HBV-DNA were quantified using quantitative real-time PCR. The positive detection rate difference of HBV-DNA, HBV-LP and Pre S1-Ag were compared, the correlation between the logarithm of HBV-DNA copies number and the absorbance value of HBV-LP was analyzed using Spearman rank correlation.@*Results@#The positive rates of HBV DNA, HBV-LP, and Pre S1-Ag were 67.22% (324/482), 73.86% (356/482), and 37.34% (180/482), respectively (P<0.01). The positive rates of the three markers were 54.57% (185/339), 64.90% (220/339), and 27.73% (94/339), respectively, in 339 HBeAg-negative CHB patients (P<0.01). In HBeAg negative patients, the positive rate of HBV-LP in 185 cases of HBV-DNA positive samples was 90.81% (168/185), which was higher than that of Pre S1-Ag with rate of 39.46% (73/185) (P<0.01).The positive rate of HBV-LP was 33.77% (52/154) in 154 cases of patients with negative HBV-DNA whose positive rate was higher than Pre S1-Ag with positive rate of 13.64% (21/154)(P<0.01). The positive rates of HBV-DNA, HBV-LP and Pre S1-Ag in HBsAg, HBeAg and HBcAb positive groups were 97.04% (131/135), 94.81% (128/135), and 60.00% (81/135), (P<0.01); The positive rates of three indexes of HBsAg, HBeAb, HBcAb positive group were 53.74% (122/227), 63.88% (145/227), and 27.31% (62/227); The positive rates of three indexes of HBsAg and HBcAb positive group were 55.79% (53/95), 67.37% (64/95), and 28.42% (27/95) (P<0.01). The absorbance value of HBV-LP was positively related with the logarithm of HBV-DNA copies number (Spearman rank correlation coefficient was 0.908, P<0.01).@*Conclusion@#There was a good correlation between HBV-LP and HBV-DNA load value, and could be used as an effective complement for the detection of HBV-DNA and HBV-M. Compared with Pre S1-Ag.

Clinical correlation between NKG2A+NK cells and regulatory T cells in patients with chronic hepatitis B virus infection / 中国免疫学杂志

Objective:To investigate the correlation between NKG2A+NK cells and regulatory T cells in peripheral blood of patients with chronic hepatitis B virus infection, and explore the clinical significances.Methods: Forty-six patients with chronic hepatitis B virus infection and 17 health individuals were included in this study.HBV DNA levels were measured by Real-time quantitative PCR ( FQ-PCR) .NKG2A+NK cells and Treg in PBMC were quantitatively analyzed by flow cytometry.Results:According to HBV DNA levels,the CHB patients were divided into two groups:Low HBV DNA group(Low viral load group,300-104 U/ml)and High HBV DNA group( High viral load group,105-108 U/ml).We found that ALT levels of High HBV DNA group were obviously higher than Low HBV DNA group(P<0.05).The frequenies of CD56dim NK cells of High HBV DNA group were obviously higher than low HBV DNA group ( P<0.05 ), and similarly the percentages of NKG2A+CD56dim NK cells of High HBV DNA group were significantly higher than Low HBV DNA and control groups ( P<0.05).We also found that the percentages of regulatory T cells ( Treg) of High HBV DNA group were significantly higher than Low HBV DNA and control groups ( P<0.05).In addition,the proportions of NKG2A+CD56dim NK cells were positively correlated with High HBV DNA levels (r=0.59,P<0.05) and the percentages of Treg(r=0.53,P<0.05) in CHB patients.Conclusion:NKG2A+CD56dim NK cells may closely relate to the HBV-related immune escape and the progress of CHB.

Study on efflux pump MexAB-OprM in carbapenem-resistant Pseudomonas aeruginosa / 中国感染与化疗杂志

Objective To study the relationship between efflux pump MexAB‐OprM and carbapenem resistance of Pseudomonas aerginosa strains .Methods The minimum inhibitory concentrations of imipenem and meropenem were determined by agar dilution method for 75 strains of P .aerginosa in the absence or presence of MC207110 to screen the phenotypes of active efflux pump .Reverse transcriptase‐polymerase chain reaction (RT‐PCR) method was used to determine the mRNA expression level of mexA which encodes the membrane fusion protein in active efflux pump MexAB‐OprM and the reference (housekeeping) gene rpoD .PCR method was used to amplify the regulatory genes mexR ,nalC ,and nalD of active efflux pump MexAB‐OprM in the strains overexpressing the efflux pump . The PCR products were subject to DNA sequencing and BLAST analysis . Results Of the 75 P .aeruginosa strains ,13 (17 .3% ) were positive for efflux pump MexAB‐OprM .Overexpression of the efflux pump was identified in 10 of the 13 strains and associated with positive regulatory genes mexR ,nalC and nalD .A Gly71→Glu mutation in nalC was found in 9 strains ,and a Ser209→Arg mutation in nalC was identified in 8 strains .Only one strain had a Thr158→Ile mutation in nalD .Eight strains had mutation in mexR .Conclusions Overexpression of multidrug efflux pump MexAB‐OprM plays an important role in carbapenem resistance of P .aeruginosa .High level expression of MexAB‐OprM is related to the mutations of its regulatory genes .

Follicular variant of papillary thyroid carcinoma:analysis of sonographic features and misdiagnosis / 中华医学超声杂志（电子版）

Objective To investigate the sonographic features of follicular variant of papillary thyroid carcinoma (FV-PTC) and to decrease misdiagnosis rate. Methods Thirty-one patients with 35 FV-PTCs and 66 patients with 75 conventional PTCs (C-PTCs) were enrolled in this study. The sonographic features were reviewed retrospectively between the two groups with universally accepted standards. Results The sonographic features of 35 FV-PTCs included irregular shapes (6/35), anteroposterior to transverse diameter ratio A/T > 1 (7/35), spiculated margins (25/35), marked hypoechogenicity (0/35), hypoechogenicity (18/35), isoechogenicity (16/35), no calcification (15/35), microcalcifications (11/35), macrocalcification (9/35), color Doppler lfow patternⅠ(20/35), color Doppler lfow patternⅡ(10/35), color Doppler lfow patternⅢ(5/35). Irregular shapes, A/T>1, spiculated margins, marked hypoechogenicity, microcalciifcations, and color type Ⅱ were rarer in FV-PTCs than in C-PTCs, while isoechogenicity, no calciifcation, macrocalciifcation, and color type Ⅲwere more frequent in FV-PTCs than in C-PTCs. The differences of the above features were statistically significant [χ2=4.276, P=0.039; χ2=8.125, P=0.004; P=0.009 (Fisher′ s exact test); χ2=8.548, P=0.003;χ2=4.898, P=0.027,χ2=7.796, P=0.005;χ2=5.462, P=0.019;P=0.001 (Fisher′s exact test)] . During the preoperative ultrasonography, 20 of 35 FV-PTCs were diagnosed as malignancy, and others were misdiagnosed as benign nodules (misdiagnosis rate was 43%). The lymphatic metastasis rate of FV-PTCs was 29%(9/31), significantly lower than C-PTCs [62%(41/66),χ2=9.246, P=0.002]. In terms of the sonographic features of metastatic lymph nodes, there was no marked difference between FV-PTCs and C-PTCs. Conclusions Some FV-PTCs are lack of malignant features, and tend to be misdiagnosed frequently when coexisting with benign thyroid nodules. Observing the echogenicity, color lfow characteristics and other features of each thyroid nodule and cervical lymph node with multiple views may decrease the misdiagnosis rate.

Study on the relationship between T cell subsets and count and virus load in patients with hepatitis C / 安徽医科大学学报

Objective To investigate the correlation between T cell subsets and HCV viralload in HCV infection. Methods Flow cytometry was used to detect peripheral T cell subsets count of 69 patients with hepatitis C and 20 cases of normal health human, and fluorescent quantitative PCR was used to detect viral load in patients with hepa-titis C. Results ① The study showed that the percentage ratio of CD4 +T cells (42.87 ±6.11) and of CD4 +/CD8 +(1.34±0.25) in patients patients was significantly lower than those of normal health group,it was 49.55±6.68 and 1.82±0.11 (P<0.01);but the percentage ratio of CD8 +T cells (32.78±5.48) was higher than that of the normal health human ( 27.35 ±4.32 ) ( P<0.01 ) . There were no significant differences between the two groups in the percentage ratio of CD3 + T cells. ② The viral load gradually increased as the percentage ratio of CD8 +T cells increased, while the percentage ratio of CD4 +T cells and the percentage ratio of CD4 +T cells and CD4 +/ CD8 + were decreased gradually. Conclusion It may be one of the important reasons as the chronic infec-tion of hepatitis C virus by change of T cell subsets, and may be an important cause leading to the decrease of T lymphocyte response ability as the expression level of HCV RNA increases.

A new function of NK cells:adaptive or memory? / 中国免疫学杂志

Recent reports have indicated that NK cells may have the function of immunological memory in common with cells of the adaptive immune system."Memory" NK cells move through the four phases of the adaptive immune response — proliferation,contraction,memory and recall which was similar to the T-cell responses."Memory" NK cells have special phenotypes.During MCMV infection,"memory" NK cells showed a greater expression of Ly49H,KLRG1,CD43 and Ly6C,and a decreased expression of CD27.In hapten (DNFB)-induced contact hypersensitivity model,the sorted liver NK cell subsets with the phenotype of Thy-1+Ly49C-I+ had the transferable memory activity,occupying around 10% of hepaedtic NK cells.These Ly49H+"memory" NK cells might produce more IFN-gamma ex vivo in response to plate-bound antibody against NK1.1 or Ly49H.Degranulation by Ly49H+ "memory" NK cells was also enhanced,as assessed by CD107a expression after ex vivo stimulation with anti-NK1.1."Memory" NK cells may move through the same migratory routes and adhesion pathways during the priming and effector phases of DNFB-primed Rag2-/-mice as conventional T cells of normal mice.Until now,it is not clear about the requirements for the triggering,regulation and maintenance of NK cell memory,but it is absolutely clear that "Memory" NK cells must play important role in host defense against pathogens.