CD8 + T cell exhaustion statuses in patients with human immunodeficiency virus infection, Mycobacterium tuberculosis infection and co-infection / 中华传染病杂志

Objective:To analyze the statuses of CD8 + T cell exhaustion in patients with human immunodeficiency virus (HIV) infection, Mycobacterium tuberculosis (MTB) infection and co-infection. Methods:A total of 87 patients infected with HIV and/or MTB in Wuxi Fifth People′s Hospital and Taicang First People′s Hospital from August 2019 to January 2020 were enrolled, including 18 cases of HIV infection, 34 cases of active tuberculosis (ATB), 19 cases of latent tuberculosis infection (LTB), seven cases of HIV coinfected with ATB, and nine cases of HIV coinfected with LTB. Another 11 healthy controls were also included. The peripheral blood of all subjects was collected for cell surface staining and intracellular cytokine staining, and flow cytometry was used to detect the expressions of activation molecules including CD62 ligand, CD44 and CD127, the transcription factor like eomesodermin (EOMES), T cell factor 1 (TCF-1), T-box expressed in T cells (T-bet), B lymphocyte-induced maturation protein 1 (Blimp-1), inhibitory receptors including programmed death-1 (PD-1) and T-cell immunoglobulin and mucin domain 3 (Tim-3) on CD8 + T cells. Mann-Whitney U test was used for statistical analysis. Results:The mean fluorescence intensities (MFIs) of the activation molecules CD62 ligand and CD44 in the HIV group were lower than those in the healthy control group, while the inhibitory receptor Tim-3 was higher than that in the healthy control group. The differences were all statistically significant ( U=31.00, 1.00 and 0.00, respectively, all P<0.010). The MFIs of CD62 ligand and CD44 in HIV coinfected with LTB group were lower than those in LTB group, while PD-1 and Tim-3 were higher than those in LTB group. The differences were all statistically significant ( U=4.00, 26.00, 6.00 and 3.00, respectively, all P<0.010). The MFIs of CD62 ligand, CD44 and CD127 in HIV coinfected with ATB group were lower than those in ATB group, while PD-1 and Tim-3 were higher than those in ATB group. The differences were all statistically significant ( U=9.00, 40.00, 45.50, 28.00 and 7.00, respectively, all P<0.010). The proportion of terminal effector CD8 + T cells in the HIV group was higher than that in the healthy control group, while the proportion of central memory CD8 + T cells was lower than that in the healthy control group. The differences were both statistically significant ( U=15.00 and 33.00, respectively, both P<0.010). The proportion of terminal effector CD8 + T cells in the HIV coinfected with LTB group was higher than the LTB group, while the proportion of central memory CD8 + T cells was lower than that in the LTB group. The differences were both statistically significant ( U=7.00 and 20.00, respectively, both P<0.010). The proportion of terminal effector CD8 + T cells in the HIV coinfected with ATB group was higher than that in ATB group, while the proportion of central memory CD8 + T cells was lower than that in ATB group. The differences were statistically significant (both U=7.00, P<0.001). The expression level of PD-1 + Tim-3 + T cells in HIV group was higher than that in healthy control group, that in HIV coinfected with LTB group was higher than that in LTB group, and that in HIV coinfected with ATB group was higher than that in ATB group. The differences were all statistically significant ( U=21.00, 6.00 and 5.50, respectively, all P<0.001). The MFI of transcription factors EOMES and TCF-1 in HIV coinfected with LTB group were lower than those in HIV group, while the MFI of T-bet was higher than that in HIV group. The differences were all statistically significant ( U=3.00, 4.00 and 9.00, respectively, all P<0.001). The MFI of EOMES and TCF-1 in HIV coinfected with ATB group were lower than those in HIV group, while the MFI of T-bet and Blimp-1 were higher than those in the HIV group. The differences were all statistically significant ( U=11.00, 14.00, 7.00 and 22.00, respectively, all P<0.050). Conclusions:MTB co-infected with HIV patients present lower immune function and a higher degree of CD8 + T cell exhaustion. In addition, HIV patients co-infected with LTB and ATB have a higher degree of CD8 + T cell exhaustion than HIV infected patients.

Expression of immunosuppressive receptor T cell immunoglobulin and immunoreceptor tyrosine-based inhibitory motif domain on CD8 + T cells in silicosis patients with Mycobacterium tuberculosis infection / 中华传染病杂志

Objective:To explore the expression and clinical significance of immunosuppressive receptor T cell immunoglobulin and immunoreceptor tyrosine-based inhibitory motif domain (TIGIT) on the peripheral blood mononuclear cells (PBMC) in silicosis patients with Mycobacterium tuberculosis infection. Methods:August 2018, a total of 78 patients with silicosis (all were quarry workers in Sanmen County, Zhejiang Province) were enrolled and divided into silicosis combined with active pulmonary tuberculosis group (APTB group), silicosis combined with latent tuberculosis infection group (LTBI group), and simple silicosis with non-tuberculosis infection group (non-TB group). Flow cytometry was used to analyze the expressions of TIGIT, programmed death-1 (PD-1) and transcription factor T-bet on PBMC from patients. Mann-Whitney U test and Pearson correlations analysis were used for statistical analysis. Results:Among the 78 patients, eight were in the APTB group, 24 in the LTBI group, and 46 in the non-TB group. The expressions of PD-1 and TIGIT on CD8 + T cells in the APTB group (29.45%(16.78%) and 65.40%(12.12%), respectively) were significantly higher than those in the LTBI group (17.40%(11.17%) and 48.30%(28.75%), respectively; U=23.500 and 43.500, respectively, P=0.000 8 and 0.020 5, respectively) and non-TB group (15.95%(12.46%) and 45.30%(19.75%), respectively; U=64.000 and 69.000, respectively, P=0.002 3 and 0.003 8, respectively), and the differences were all statistically significant. The expression of TIGIT was positively correlated with PD-1 on CD8 + T cells in silicosis patients ( r=0.434 3, P<0.01). The proportion of PD-1 + TIGIT + CD8 + T cells in the APTB group (19.90%(22.67%)) was significantly higher than those in the non-TB group (11.55%(11.29%), U=76.500, P=0.007 1) and LTBI group (11.55%(10.53%), U=41.000, P=0.015 4), while the proportion of PD-1 -TIGIT -CD8 + T cells in the APTB group (30.60%(12.90%)) was significantly lower than non-TB group (48.90%(18.98%), U=58.000, P=0.001 3) and LTBI group (47.20%(24.59%), U=41.000, P=0.015 4). The differences were all statistically significant. The expression of T-bet on the peripheral blood CD8 + T cells in the APTB group (29.45%(16.78%)) was higher than that in the non-TB group (15.95%(12.46%)) and the LTBI group (17.40%(11.17%)), and the differences were both statistically significant ( U=46.500 and 46.000, respectively, P=0.000 3 and 0.028 3, respectively). The expression of T-bet on CD8 + T cells was positively correlated with TIGIT on CD8 + T cells ( r=0.456 7, P<0.01). The expression of T-bet on PD-1 + TIGIT + CD8 + T cells in the APTB group (65.40%(12.12%)) was higher than those in the LTBI group (48.30%(28.75%), U=23.500, P=0.000 8) and non-TB group (45.30%(19.75%), U=65.000, P=0.002 6), and the differences were both statistically significant. Conclusion:The immunosuppressive receptor PD-1 and TIGIT are highly expressed on CD8 + T cells in silicosis patients with active pulmonary tuberculosis, which indicates CD8 + T cells exhaustion in these population, while the highly co-expression of T-bet suggests the exhausted subsets may have reversed potentiality.

Effects of lncRNA CDKN2B-AS1 targeting miR-98-5p on proliferation and invasion of lung cancer A549 cells / 中华内分泌外科杂志

Objective:To investigate the effects of long non-coding RNA (lncRNA) CDKN2B-AS1 targeting miR-98-5p on proliferation and invasion of lung cancer A549 cells.Methods:A549 cells cultured in vitro were divided into control group, pcDNA group (transfected with pcDNA) , CDKN2B-AS1 group (transfected with pcDNA CDKN2B-AS1) and double transfection group (transfected with pcDNA CDKN2B-AS1 and pcDNA miR-98-5p) . The expression of lncRNA CDKN2B-AS1, miR-98-5p and the protein expression of PCNA, MMP-9 in A549 cells were detected. The activity, clone number, cloning efficiency, and the number of invasive cells of A549 cells were detected.Results:Compared with pcDNA group, the expression level of lncRNA CDKN2B-AS1 [ (2.14±0.14) vs (1.03±0.10) ], OD value in each time points, clone number [ (314.60±18.13) vs (220.08±12.46) ], cloning efficiency [ (85.81±3.06) % vs (60.03±2.85) %], invasive cell number [ (233.30±18.98) vs (140.84±12.30) ], expression levels of PCNA [ (0.78±0.08) vs (0.48±0.07) ] and MMP-9 [ (0.75±0.06) vs (0.38±0.06) ] proteins in A549 cells in CDKN2B-AS1 group were significantly increased ( P<0.05) ; the expression level of miR-98-5p [ (0.23±0.03) vs (0.99±0.09) ] was significantly decreased ( P<0.05) ; compared with CDKN2B-AS1 group, there was no significant difference in the expression level of lncRNA CDKN2B-AS1 in A549 cells in double transfection group ( P>0.05) , while the expression level of miR-98-5p in A549 cells was significantly increased ( P<0.05) . The OD value in each time points, clone number, cloning efficiency, invasive cell number, expression levels of PCNA and MMP-9 proteins were significantly decreased ( P<0.05) . Conclusion:LncRNA CDKN2B-AS1 can promote the proliferation and invasion of lung cancer A549 cells by targetingly inhibiting the expression of miR-98-5p.

Diagnostic values of interleukin-22, interferon-γ and macrophage migration inhibition factor in pleural effusion for tuberculous pleurisy / 中华传染病杂志

Objective:To investigate the diagnostic values of interleukin-22 (IL-22), interferon-γ(IFN-γ)and macrophage migration inhibition factor (MIF) in pleural effusion for tuberculosis pleurisy.Methods:From April 2018 to May 2019, a total of 77 patients including 45 cases of tuberculous pleurisy, 19 cases of malignant pleurisy, 13 cases of parapneumonia and 13 cases of healthy control in Wuxi Fifth People′s Hospital were enrolled. The levels of IL-22, IFN-γ and MIF in plasma and pleural effusion were detected by enzyme linked immunosorbent assay (ELISA). Mann-Whitney U test was used for statistical analysis.The receiver operating characteristic (ROC) curve was used to evaluate the diagnostic values of IL-22, IFN-γ and MIF for tuberculous pleurisy. Results:The median levels of IL-22, IFN-γ, MIF and adenosine deaminase in 45 cases with pleural effusion in tuberculosis pleurisy group were 396.8 ng/L, 2 200.0 ng/L, 241.3 μg/L and 70.8 U/L, respectively, which were all significantly higher than 32 cases with non-tuberculosis pleurisy group, including 19 cases with malignant pleurisy and 13 cases with parapneumonia (52.8 ng/L, 232.3 ng/L, 179.6 μg/L and 17.0 U/L, respectively). The differences were all statistically significant ( U=179.000, 118.500, 287.000, 162.000, respectively, all P<0.05). The median levels of IL-22 and IFN-γ in plasma of tuberculosis pleurisy group were 20.0 ng/L and 45.9 ng/L, respectively, which were both higher than healthy control group (14.3 ng/L and 33.4 ng/L, respectively). The level of MIF was 96.2 μg/L, which was lower than healthy control (159.5 μg/L). The differences were all statistically significant ( U=74.000, 13.000 and 73.000, respectively, all P<0.05). The areas under ROC curve (AUC) of IL-22, IFN-γ and MIF in pleural effusion for the diagnosis of tuberculosis pleurisy were 0.876, 0.917 and 0.682, respectively.The sensitivities were 93.75%, 100.00% and 63.64%, respectively; the specificities were 82.22%, 91.11% and 65.85%, respectively. The median levels of IL-22 and IFN-γ in plasma in tuberculosis pleurisy group at two months of follow-up after anti-tuberculosis therapy were 16.0 ng/L and 33.9 ng/L, respectively, which were both lower than baseline (20.0 ng/L and 44.7 ng/L, respectively). The differences were both statistically significant ( U=2.156 and 2.221, respectively, both P<0.05). Conclusion:IFN-γ and IL-22 in pleural effusion could be used as effective indicators to identify tuberculous pleurisy, and the dynamic monitoring of IL-22 in patients′plasma could be an important biomarker in evaluating the efficacy of anti-tuberculosis treatment.

Preliminary study on predictive indicators of disease severity in patients with influenza A (H1N1) / 中华传染病杂志

Objective:To analyze the differences of peripheral blood transcriptome between mild and severe influenza A (H1N1) patients, and to find indicators for the assessment of disease severity.Methods:A total of ten patients (five patients with mild disease and five patients with severe disease) diagnosed with H1N1 infection from January to May 2018 at Huashan Hospital, Fudan University in Shanghai were enrolled, and five healthy people were also enrolled as controls. The peripheral blood of patients was collected for transcriptome sequencing at the time when they were first diagnosed. Measurement data were compared using t test or Mann-Whitney U test. The count data were compared using Fisher exact test when appropriate. Data analysis of transcriptome predictions was performed using bioinformatics methods. Results:The platelet counts were significantly different between mild and severe groups ((163.4±21.5 )×10 9/L vs (255.6±52.5)×10 9/L, t=3.636, P=0.007). There were no differences between the two groups in gender, age, white blood cell counts, neutrophil percentage, lymphocyte percentage and hemoglobin levels (all P>0.05). However, the average expression levels of matrix metalloproteinase (MMP) 8 and MMP9 in severe group (18.41 and 174.00, respectively) were both higher than those in mild group (2.33 and 22.91, respectively) and healthy control (1.43 and 34.65, respectively; all P<0.01). Conclusion:MMP8 and MMP9 could be expected to serve as the molecular biological markers for predicting the disease severity in patients with influenza A (H1N1) infection.

Clinical characteristics of patients with positive blood culture for Mycobacterium tuberculosis in a general hospital / 中华传染病杂志

Objective To retrospectively analyze the clinical and laboratory characteristics of patients with positive blood culture results for Mycobacterium tuberculosis (M.tb).Methods The clinical laboratory database of patients suspected with disseminated tuberculosis from January 2009 to January 2017 in Huashan Hospital affiliated with Fudan University were collected and analyzed.The clinical manifestations,laboratory characteristics and outcomes between disseminated tuberculosis patients with positive blood culture (positive blood culture group) for M.tb and negative results (negative blood culture group) were compared.T test,Mann-Whitney U test and Fisher exact test were used for statistical analysis.Results A total of 5 589 patients suspected with M.tb infection had peripheral blood culture for mycobacterium.Positive blood culture for M.tb was found in 26 disseminated tuberculosis patients,while 6 patients finally identified as nontuberculous mycobacterium (NTM) with species identification,and 22 disseminated tuberculosis patients with negative blood culture results were enrolled during the same period as control.The mean age ([49.1 ± 10.1] years old vs [38.3 ± 17.1] years old,t =2.460,P =0.018),the proportion of diagnosed with fever of unknown origin at admission (FUO) (65.0％ [13/20] vs 13.6％ [3/22],P =0.001),the proportion of diagnosed with focal infection (30.0％ [6/20] vs 86.4％ [19/22],P =0.001),the proportion of patients with other diseases (75.0％[15/20] vs 22.7％ [5/22],P =0.002),the proportion of patients with hematological diseases (35.0％ [7/20] vs 4.5％ [1/22],P =0.018) and the proportion of patients with tumor (20％ [4/20] vs 0[0/22],P =0.043) in the positive blood culture group were significantly different from those in the negative blood culture group.Laboratory examinations of the percentage of neutrophils,the percentage of lymphocytes,the percentage of monocytes,the value of neutrophil/lymphocyte,the level of hemoglobin,the level of erythrocyte sedimentation rate,the level of C-reactive protein,the level of procalcitonin and the positive rate of T-SPOT.TB in positive blood culture groups were significantly different from those in negative blood culture group (all P ＜ 0.05).Conclusions Peripheral blood M.tb culture is more likely to be positive for those elder disseminated tuberculosis patients with hematological diseases or tumors,and those with increase of neutrophil counts and inflammation markers but reduction of lymphocyte counts and hemoglobin.

Effect of betulinic acid on spatial learning and memory in type 2 diabetic rats and its mechanism / 中华行为医学与脑科学杂志

Objective To investigate the effects of betulinic acid on spatial learning and memory in rats with type 2 diabetes mellitus(T2DM) and to explore its underlying mechanisms.Methods Forty-five male Wistar rats were randomly divided into control group (Sham),T2DM group and betulinic acid treatment group (BA) with 15 in each group.The rats in the latter two groups were given a high fat diet plus low dose of streptozotocin (STZ) to induce T2DM,and the rats in BA group was given 10 mg/kg BA for intragastric administration once daily for 12 weeks,the rats in T2DM group and Sham group were given an equal volume of physiological saline.Morris water maze method was used to detect the cognitive function.The mRNA levels of synaptic plasticity related protein GAP-43,SYP and PSD-95 were detected by RT-PCR.The protein expressions of cholinergic neurotransmitter ChAT and Ach were detected by Elisa.Results Compared with the Sham group,the escape latency of the T2DM group was significantly prolonged on days 2-4 ((62.28 ± 2.97)s vs (47.09±2.16)s;(48.52±3.09) s vs (26.18±2.21)s;(42.31±2.80)s vs (13.42±1.23)s),the original platform quadrant time was significantly reduced ((24.60±2.42) s vs (41.85 ± 1.98) s),GAP-43,SYP,ChAT and Ach levels were significantly lower,the differences were statistically significant (the t values were 16.37,22.34,39.78,20.42,116.01,91.35,71.84,21.88 and 21.11,respectively,all P＜ 0.05).Compared with T2DM group,the escape latency of BA group in 2-4 days were significantly shortened ((55.61±2.75)s vs (62.28±2.97)s;(31.35±2.63)s vs (48.52±3.09)s;(42.31±2.80)s vs (16.58± 1.37) s),the original platform quadrant time increased significantly ((37.58± 2.31) s vs (24.60±2.42) s),and GAP-43,SYP,PSD-95,ChAT and Ach levels were significantly higher,and the differences were statistically significant (the t values were 6.80,16.21,33.54,14.55,41.83,35.23,36.20,12.82 and 9.97,respectively,all P＜0.05).Conclusion Betulinic acid can improve the spatial learning and memory of type 2 diabetic rats,and increasing the expression of GAP-43,SYP,PSD-95,ChAT and Ach may be its potential mechanism.

The effect of 3-n-butylphthalide on learning and memory abilities in vascular dementia rats / 中华行为医学与脑科学杂志

Objective To investigate the effect of 3-n-butylphthalide (NBP) on learning and memory abilities in vascular demensia (VD) rats. Methods VD model rats was established by two-vessel method. 60 3-month-old Wistar male rats were randomly divided into VD group, sham-operate group and NBP group. Rats in NBP group were given NBP 120 mg · kg-1 · d-1 ,VD and sham-operate group were given equal quality vegetable oil.Morris water maze was used to assess the learning and memory abilities in each group and HE staining was used to observe the hippocampus morphology of the rats. Results The escape latency in hidden plat test was( (38.34 ±2.46 ) s, ( 14.83 ± 3.77s ), ( 75.74 ± 6.33 ) s ) and the original platform quadrant time was ( ( 26.45 ± 4.66 ) s,(35.21 ±3.78)s, ( 18.67 ±5.39)s) in NBP group,sham-operate group and VD group respectively. Compared to VD group, NBP group had obviously decreased escape latency in hidden plat test, increased original platform quadrant time and distinctly decreased the necrosis of the neurons in HE dyeing. Conclusion NBP can improve the learning and memory deficits in VD rats.