Early diagnosis value of peripheral blood PPARγ combined with IFN-γ detection for tuberculosis infection in patients with end-stage renal disease / 中国热带医学

@#Abstract: Objective　To explore the early diagnostic value of peripheral blood peroxisome proliferator-activated receptor γ (PPARγ) combined with γ-interferon (IFN-γ) release assay (IGRA) in the diagnosis of pulmonary tuberculosis in patients with end-stage renal disease (ESRD), and to provide reference for clinical diagnosis and treatment. Methods　From January 2019 to December 2021, 70 ESRD patients with suspicious symptoms of pulmonary tuberculosis were treated at Hebei Chest Hospital were selected as the research objects. According to the examination results, they were divided into ESRD group (40 cases) and ESRD complicated by pulmonary tuberculosis (40 cases, comorbidity group). In addition, 40 cases with pulmonary tuberculosis were used as the PTB group. All three groups of patients underwent IGRA test, and the peripheral blood PPARγ level was detected by enzyme-linked immunosorbent assay, and the diagnostic value of PPARγ combined with IGRA test for ESRD patients with pulmonary tuberculosis was explored. Results The expression level of PPARγ and IFN-γ content in the PTB group and the comorbidity group were obviously higher than those in the ESRD group (P<0.05), while the differences in PPARγ expression level and IFN-γ content between the PTB and comorbidity groups were not statistically significant (P>0.05). The ROC curve showed that the areas under the curve (AUC) of PPARγ and IGRA in the diagnosis of end-stage renal disease combined with tuberculosis were 0.823 (95%CI: 0.722-0.925) and 0.773 (95%CI: 0.662-0.883), respectively, and the AUC of combined detection was 0.928 (95%CI: 0.871-0.984), which was better than that of PPARγ and IGRA alone (Z/P=2.057/0.039, 2.843/0.005). The Kappa values of serum PPARγ and IGRA test compared with the clinical gold standard results in the diagnosis of ESRD complicated with pulmonary tuberculosis were 0.557 and 0.444 (P<0.05). The combined screening of ESRD with pulmonary tuberculosis was consistent with the clinical gold standard (Kappa=0.661, P<0.05). Among the 30 ESRD patients complicated with pulmonary tuberculosis, the sensitivity of PPARγ combined with IGRA test in diagnosis of ESRD complicated with pulmonary tuberculosis was 93.33% (28/30), which was higher than 70.00% (21/30) of PPARγ and 66.67% (20/30) of IGRA test alone (P<0.05). Conclusions Peripheral blood PPARγ and IGRA tests have certain diagnostic value for ESRD complicated with tuberculosis, and the combined detection of the two can improve the sensitivity and reduce the rate of missed diagnosis, which is worthy of clinical promotion.

Regulation of microRNA-126 on the polarization of human macrophages stimulated by Porphyromonas gingivalis lipopolysaccharide / 中华口腔医学杂志

Objective: To study the effect of microRNA-126 (miR-126) on the polarization of human monocyte-derived macrophages stimulated by Porphyromonas gingivalis (Pg) lipopolysaccharide (LPS). Methods: Macrophages derived from human myeloid leukemia mononuclear cells were stimulated by Pg-LPS (5 mg/L) and by Pg-LPS (5 mg/L) after 24 h-transfection of miR-126 mimic or negative control RNA for 48 h, respectively. Real-time quantitative-PCR (qRT-PCR), enzyme-linked immunosorbent assay (ELISA) and Western blotting were conducted to detect the changes in miR-126, pro-inflammatory factor tumor necrosis factor-α (TNF-α), anti-inflammatory factors interleukin-10 (IL-10), inducible nitric oxide synthase (iNOS), arginase-1 (Arg-1) and M1 polarization-related pathways such as nuclear factor kappa-B (NF-κB) and mitogen-activated protein kinase (MAPK) signaling pathways. Results: Compared with non-LPS stimulation group (TNF-α: 1.000±0.020, iNOS: 1.125±0.064, miR-126: 1.004±0.113, IL-10: 1.003±0.053, Arg-1: 1.130±0.061), the mRNA levels of TNF-α (3.105±0.278) and iNOS (4.296±0.003) increased significantly (t=6.53, P=0.003; t=42.63, P<0.001, respectively), while miR-126, IL-10 and Arg-1 expressions (0.451±0.038, 0.545±0.004 and 0.253±0.017) decreased significantly (t=7.95, P=0.001; t=7.36, P=0.002; t=11.94, P<0.001, respectively) after Pg-LPS stimulated by human-derived macrophages for 48 h. The protein expression of iNOS, TNF-α, Arg-1 and IL-10 were consistent at mRNA levels. Meanwhile, the expressions of phospho-NF-κB p65 (p-p65), phospho-extracellular signal-regulated kinase (p-ERK) and phospho-p38 MAPK (p-p38) increased significantly, while the expression of Arg-1 decreased significantly. Compared with the negative controls (scramble RNA) (TNF-α: 1.141±0.197, iNOS: 1.173±0.115, IL-10: 1.032±0.138, Arg-1: 0.933±0.044), the mRNA levels of TNF-α (0.342±0.022) and iNOS (0.588±0.085) expressions significantly decreased (t=5.35, P=0.006; t=5.05, P=0.007), while IL-10 (1.786±0.221) and Arg-1 expressions (2.152±0.229) significantly increased (t=3.71, P=0.021; t=6.21, P=0.003) after Pg-LPS stimulation with miR-126 mimic transfection. The relative protein expressions of iNOS, p-p65, p-ERK and p-p38 significantly decreased (t=13.00, P<0.001; t=6.98, P=0.002; t=10.86, P<0.001; t=8.32, P=0.001), while the protein level of Arg-1 significantly increased (t=12.08, P<0.001). Conclusions: Pg-LPS could promote M1 polarization of macrophages. miR-126 might inhibit the effect of Pg-LPS on the M1 polarization of macrophages through down-regulating NF-κB and MAPK signaling pathways.

Application and effect of virtual-reality surgery simulation system in minimally invasive cataract surgery training for ophthalmology residents / 国际眼科杂志(Guoji Yanke Zazhi)

@#AIM: To investigate the application and effect of virtual-reality surgery exercise in minimally invasive cataract surgery training for ophthalmology residents.<p>METHODS:Twenty ophthalmology residents with equal seniority who had completed 3a standardized residency training in the Affiliated Eye Hospital of Nanjing Medical University from 2019 to 2021 were prospectively enrolled. After passing the theoretical examination, residents were randomly divided into virtual surgery exercise(Dry-lab)group(<i>n</i>=10)and real animal surgery exercise(Wet-lab)group(<i>n</i>=10). Dry-lab and Wet-lab group residents performed training using the Eye SI surgical simulator and pig eye respectively. At the end of the training, the overall training effects of the residents in both groups were rated using the Eye SI surgical simulator and the real pig eye operation, and the module training effects of the residents in both groups were rated using the virtual surgical simulator. Furthermore, a questionnaire survey was used to objectively evaluate the two training methods.<p>RESULTS:Residents in Dry-lab group had significantly higher total scores on both operation assessments,simulator assessment and real pig eye operation assessment than Wet-lab group(88.03±1.34 <i>vs</i> 80.35±2.87, 87.50±3.03 <i>vs</i> 77.60±5.62, 88.57±1.89 <i>vs</i> 83.10±3.22, all <i>P</i><0.01). The simulator module assessment results showed that the residents in Dry-lab group scored significantly better than Wet-lab group in terms of scores and completion time on each module(<i>P</i><0.01). The questionnaire results showed that Dry-lab group rated better than Wet-lab group in terms of the novelty of training, the proximity to the real surgical experience, the degree of help to the improvement of microsurgery skills, the confidence to perform real surgery, and the overall satisfaction of surgical training(<i>P</i><0.05). <p>CONCLUSION:Applying virtual-reality surgery exercise to cataract surgery skills training for ophthalmology residents can significantly improve the cataract skills, increase overall training satisfaction, and help residents enhance their confidence, psychological quality, decision-making, and processing ability during real surgery at the initial stage of practice. This provides a new standard and model for establishing a formal and standardized cataract surgery training system for ophthalmology residents.

Evaluation of Autofluorescence in Identifying Parathyroid Glands by Measuring Parathyroid Hormone in Fine-Needle Biopsy Washings.

Background: Near-infrared autofluorescence imaging has potentially great value for assisting endocrine surgeons in identifying parathyroid glands and may dramatically change the surgical strategy of endocrine surgeons in thyroid surgery. This study is designed to objectively evaluate the role of near-infrared autofluorescence imaging in identifying parathyroid glands during thyroid surgery by measuring intraoperative parathyroid hormone in fine-needle aspiration biopsy washings. Methods: This study was conducted at a tertiary referral teaching hospital in China from February 2020 to June 2020. Patients undergoing total thyroidectomy with or without neck lymph node dissection were consecutively included. The surgeon used near-infrared autofluorescence imaging to identify parathyroid glands during thyroid surgery and confirmed suspicious parathyroid tissues by measuring their intraoperative parathyroid hormone. Nanocarbon was injected into the thyroid gland if the thyroid autofluorescence intensity was too strong. The sensitivity and accuracy of near-infrared autofluorescence imaging and vision for identifying parathyroid glands, and the difference in autofluorescence intensity in various tissues were the main outcomes. Results: Overall, 238 patients completed the trial. Based on the pathological and aIOPTH results, the sensitivity of near-infrared autofluorescence imaging for detecting parathyroid glands (568 of 596 parathyroid glands; 95.30%)was significantly higher than that of vision (517 of 596 parathyroid glands; 86.74%, P<.001). The accuracy of near-infrared autofluorescence imaging (764 of 841 tissues; 90.84%) was significantly higher than that of vision (567 of 841 tissues; 67.42%, P<.001) when the evaluations of certain tissues were inconsistent. There was a significant difference between the autofluorescence intensity of the parathyroid glands and that of the lymph nodes (74.19 ± 17.82 vs 33.97 ± 10.64, P<.001). Conclusion: The use of near-infrared autofluorescence imaging, along with intraoperative parathyroid hormone and nanocarbon for the identification of parathyroid glands in thyroid surgery may increase the number of confirmed parathyroid glands. Using near-infrared autofluorescence imaging can effectively distinguish lymph nodes and parathyroid glands during lymph node dissection.

CD137 agonist induces gastric cancer cell apoptosis by enhancing the functions of CD8+ T cells via NF-κB signaling.

BACKGROUND: CD137 is a target for tumor immunotherapy. However, the role of CD137 in gastric cancer (GC), especially in inducing GC cell apoptosis, has not been studied. METHODS: Foxp3+ and CD8+ T cells in GCs were investigated using immunohistochemistry (IHC). CD137 expression in GCs was detected using flow cytometry, IHC and immunofluorescence (IF). Peripheral blood mononuclear cells (PBMCs) and CD8+ T cells isolated from peripheral blood were stimulated with a CD137 agonist in vitro. CD8+ T cell proliferation and p65 expression was examined using flow cytometry. P65 nuclear translocation was analyzed using IF. IL-10, TGF-ß, IFN-γ, perforin and granzyme B were detected using real-time quantitative PCR (real-time PCR). PBMCs and primary GC cells were cocultured and stimulated with a CD137 agonist in vitro. Apoptosis of primary GC cells was detected using flow cytometry. RESULTS: Our data demonstrated that GC tumors showed characteristics of an immunosuppressive microenvironment. CD137 was predominantly expressed in CD8+ T cells in GCs and had a positive correlation with tumor cell differentiation. The CD137 agonist promoted CD8+ T cell proliferation and increased the secretion of IFN-γ, perforin and granzyme B, which induced primary GC cell apoptosis. Mechanistically, this study found that the CD137 agonist induced NF-κB nuclear translocation in CD8+ T cells. CONCLUSION: Our results demonstrated that a CD137 agonist induced primary GC cell apoptosis by enhancing CD8+ T cells via activation of NF-κB signaling.

Follow-up study of the pulmonary function and related physiological characteristics of COVID-19 survivors three months after recovery.

BACKGROUND: The long-term pulmonary function and related physiological characteristics of COVID-19 survivors have not been studied in depth, thus many aspects are not understood. METHODS: COVID-19 survivors were recruited for high resolution computed tomography (HRCT) of the thorax, lung function and serum levels of SARS-CoV-2 IgG antibody tests 3 months after discharge. The relationship between the clinical characteristics and the pulmonary function or CT scores were investigated. FINDINGS: Fifty-five recovered patients participated in this study. SARS-CoV-2 infection related symptoms were detected in 35 of them and different degrees of radiological abnormalities were detected in 39 patients. Urea nitrogen concentration at admission was associated with the presence of CT abnormalities (P = 0.046, OR 7.149, 95% CI 1.038 to 49.216). Lung function abnormalities were detected in 14 patients and the measurement of D-dimer levels at admission may be useful for prediction of impaired diffusion defect (P = 0.031, OR 1.066, 95% CI 1.006 to 1.129). Of all the subjects, 47 of 55 patients tested positive for SARS-CoV-2 IgG in serum, among which the generation of Immunoglobulin G (IgG) antibody in female patients was stronger than male patients in infection rehabilitation phase. INTERPRETATION: Radiological and physiological abnormalities were still found in a considerable proportion of COVID-19 survivors without critical cases 3 months after discharge. Higher level of D-dimer on admission could effectively predict impaired DLCO after 3 months discharge. It is necessary to follow up the COVID-19 patients to appropriately manage any persistent or emerging long-term sequelae. FUNDING: Key Scientific Research Projects of Henan Higher Education Institutions.

Effect of Ginsenoside Rg1 on Lipid Uptake and Oxidation in HepG2 Cells of Nonalcoholic Fatty Liver Disease / 中国实验方剂学杂志

Objective:To investigate the effect and mechanism of ginsenoside Rg1（G-Rg1）in ameliorating lipid uptake and oxidation in HepG2 cells induced by free fatty acids (FFA). Method:HepG2 cells were divided into normal group, model group，low-dose ginsenoside Rg1 group (25 μmol·L-1) and high-dose G-Rg1 group (50 μmol·L-1). HepG2 cells were treated with 1 mmol·L-1 free fatty acid for 24 h to construct the NAFLD cell model, and then treated with 25，50 μmol·L-1 G-Rg1 for 24 h. The effect of G-Rg1 on HepG2 cell activity was determined by cell counting kit-8（CCK-8） assay. The level of triglyceride (TG) was detected by micro method. The accumulation of lipid droplets was observed by oil red O staining. Quantitative real-time fluorescence polymerase chain reaction (Real-time PCR) and Western blot were used to detect the alterations of key genes and proteins relating to lipid uptake and metabolism. Result:Compared with the normal group, the intracellular TG level and the absorbance of the oil red O staining in the model group were significantly increased (P<0.01). Compared with the model group, G-Rg1 reduced TG and lipid deposition were significantly reduced (P<0.01).Results of Real-time PCR and Western blot showed that compared with normal group, model group peroxisome proliferators-activated receptors gamma（PPARγ），fatty acid binding protein 1（FABP1），fatty acid transport protein 2/5（FATP2/5）and fatty acid translocase（CD36）expressions increased（P<0.05），whereas peroxisome proliferators-activated receptors α（PPARα），carnitine palmitoyltransferase 1（CPT1）and peroxisomal acyl-coenzyme A oxidase 1（ACOX1）expressions decreased（P<0.05）. Compared with the model group, the expressions of PPARγ, FABP1, FATP2, FATP5 and CD36 in the G-Rg1 group were decreased (P<0.05，P<0.01), while the expressions of PPARα, CPT1 and ACOX1 were increased (P<0.05，P<0.01). Conclusion:G-Rg1 can ameliorate lipid deposition in NAFLD cell model by reducing lipid uptake and increasing lipid oxidation.

Polyethylene glycol and divalent salt-induced DNA reentrant condensation revealed by single molecule measurements.

In this study, we investigated the DNA condensation induced by polyethylene glycol (PEG) with different molecular weights (PEG 600 and PEG 6000) in the presence of NaCl or MgCl2 by using magnetic tweezers (MT) and atomic force microscopy (AFM). The MT measurements show that with increasing NaCl concentration, the critical condensation force in the PEG 600-DNA or PEG 6000-DNA system increased approximately linearly. PEG 6000 solution has a larger critical force than PEG 600 solution at a given NaCl concentration. In comparison, a parabolic trend of the critical condensation force was observed with increasing MgCl2 concentration, indicating that DNA undergoes a reentrant condensation. The AFM results show that the morphologies of the compacted DNA-PEG complexes depended on the salt concentration and were consistent with the MT results.

Comparison of cardiac function parameters in gated myocardial SPECT determined by filtered backprojection and OSEM reconstruction methods / 中华核医学杂志

Objective To compare the cardiac function parameters in gated SPECT determined by filtered back projection (FBP) and OSEM reconstruction methods. Methods One hundred and forty-four patients underwent 99Tcm-MIBI gated-SPECT imaging studies. The parameters LVEF, EDV and ESV, were derived using quantitative gated SPECT (QGS), four-dimensional model SPECT (4D-MSPECT) and emory cardiac toolbox (ECToolbox) softwares. Each image was reconstructed by FBP or OSEM. Bland-Altman analysis and paired t-test were applied to evaluate those parameters. Results Correlation coefficients for LVEF, EDV and ESV between FBP and OSEM methods were all more than 0.93 (all P<0.001). EDV calculated by FBP was lower than that by OSEM using QGS software, but became the opposite when using 4D-MSPECT and ECToolbox softwares. (QGS: (82.2±39.1) ml vs (83.5±40.8) ml, t=-2.53, P<0.05; 4D-MSPECT: (93.5±46.9) ml vs (88.8±45.2) ml, t=5.95, P<0.01; ECToolbox: (106.4±51.1) ml vs (100.8±49.0) ml, t=3.99, P<0.01). ESV calculated by FBP was higher than that by OSEM using 4D-MSPECT software (4D-MSPECT:(37.5±41.4) ml vs (34.8±37.6) ml, t=3.92, P<0.01). LVEF calculated by FBP was lower than that by OSEM using QGS software ((62.1±16.9)% vs (63.1±16.1)%, t=-3.14, P<0.05), but higher than that by OSEM using ECToolbox software ((74.1±18.8)% vs (71.3±17.1)%, t=5.28, P<0.01). Conclusion Generally, cardiac functional parameters based on FBP and OSEM construction methods correlated well, although they might have singnificantly different results.

18F-FDG/99Tcm-MIBI SPECT myocardial imaging for the detection of myocardial viability in patients with acute myocardial infarction / 中华核医学杂志

Objective To investigate the value of 18F-FDG/99Tcm-MIBI SPECT myocardial imaging for the detection of myocardial viability and prognosis in patients with AMI. Methods 18F-FDG/99Tcm-MIBI SPECT myocardial imaging was performed in 98 consecutive patients [man 87, women 11; average age (58 ±11)y] with AMI. The myocardium was scored individually for nine segments: mildly decreased uptake = 1,significantly decreased uptake = 2, and no uptake = 3. Perfusion defect but preserved 18 F-FDG uptake was defined as perfusion-metabolism mismatch, indicating jeopardized but viable myocardium. Perfusion defect and decreased 18 F-FDG uptake were defined as match, indicating myocardial necrosis. Echocardiogram was performed before and after treatment for evaluating the LVEF. All patients were followed after treatment.The rate of cardiac events was calculated and compared between patients with medication and revascularization. Paired t test, Chi-square test and log-rank test were used for statistical analysis. Results In the group with viable myocardium, 27 patients received revascularization and 10 received medication. In the group with infarcted myocardium, 26 patients received medication and 35 received revascularization. Patients underwent revascularization and with medication had no significant difference in improvement of LVEF between both groups (viable myocardium group: χ2 = 0.509, P > 0. 05; infarcted myocardium group: χ2 =0.035, P > 0.05). In viable myocardium group, cardiac event rate was significantly higher in patients with medication than in those who had undergone revascularization (50.0% vs 14.8%, χ2 =4.91, P<0.05).In the infarcted myocardium group, cardiac event rate was also significantly higher in patients with medication (30.7% vs5.7% ,χ2 =6.83, P<0.05). Conclusions 18F-FDG/ -MIBI SPECT myocardial imaging may well be of value but limited for the detection of myocardial viability and prediction of improvement in cardiac function as well as prognosis. However, more prospective data are needed for final evaluation.