Interferon-gamma release assays as a tool for differential diagnosis of gastrointestinal tuberculosis

In this editorial,we comment on an article published in a recent issue of the World Journal of Clinical Cases.There is a pressing need for reliable tools for diagnosing tuberculosis(TB)of the gastrointestinal tract.Despite advancements in the diagnosis and treatment,TB remains a global health challenge.Ali et al demon-strated that TB may mimic gastrointestinal conditions,such as gastric outlet obstruction,causing a delay in the diagnosis.Furthermore,the latter complication is frequently observed during infections,including Helicobacter pylori,and rarely is related to TB,as in the presented case.In line with this,we think that laboratory tests based on interferon-gamma release assays can be a helpful tool for diagnosing latent TB paced in the gastrointestinal tract.Innovative strategies and approaches for diagnosing latent/active extra pulmonary TB are crucial for establishing the diagnosis early and enhancing treatment strategies to mitigate the global burden of TB.

Deregulation of interferon-gamma receptor 1 expression and its implications for lung adenocarcinoma progression

Interferon-gamma(IFN-γ)plays a dual role in cancer;it is both a pro-and an antitumorigenic cytokine,depending on the type of cancer.The deregulation of the IFN-γcanonic pathway is associated with several disorders,including vulner-ability to viral infections,inflammation,and cancer progression.In particular,the interplay between lung adenocarcinoma(LUAD)and viral infections appears to exist in association with the deregulation of IFN-γsignaling.In this mini-review,we investigated the status of the IFN-γsignaling pathway and the expression level of its components in LUAD.Interestingly,a reduction in IFNGR1 expression seems to be associated with LUAD progression,affecting defenses against viruses such as severe acute respiratory syndrome coronavirus 2.In addition,alterations in the expression of IFNGR1 may inhibit the antiproliferative action of IFN-γsignaling in LUAD.

Evaluation of Interferon-Gamma Release Assay Testing and Tuberculin Skin Test for Early Diagnosis of Tuberculosis in Children and Adolescents

Background: This study aimed to evaluate the diagnostic value of interferon-γ release assay (IGRA), a sensitive microbiological diagnostic method, in children and adolescents with suspected tuberculosis in a country with a high burden of tuberculosis. Method: This study included 581 children and adolescents aged 4 - 19 years who were suspected of having tuberculosis, were latently infected with Mycobacterium tuberculosis, and had received at least one dose of BCG vaccine between April 17, 2019, and February 24, 2021. The study evaluated the TST results of 106 patients who had a positive Quantiferon test and were suspected of having tuberculosis. Results: The study included 581 patients aged between 4 and 19 years. Of these, 106 patients tested positive for the Quantiferon test, while 19 were indeterminate and 456 were negative. The Quantiferon test positivity rate was 18.24%. Among the 106 QFT-Plus-positive cases, 23 patients also tested positive for TST. The difference in distribution was found to be statistically significant. Conclusion: The QFT-Plus test is considered an alternative to TST and other microbiological diagnostic methods for early tuberculosis diagnosis, particularly in children and adolescents.

Modulation of the activation of Statl by the interferon-gamma receptor complex

The activation of Statl by the interferon-gamma （IFN-γ） receptor complex is responsible for the transcription of a significant portion of IFN-γ induced genes. Many of these genes are responsible for the induction of an apoptotic state in response to IFN-γ. In the absence of Stat 1 activation, IFN-γ instead induces a proliferative response. Modifying Stat 1 activation by IFN-γ may have pharmacological benefits. We report that the rate of activation of Statl can be altered in HeLa cells by overexpressing either the IFN-γ R1 chain or the IFN-γ R2 chain. These alterations occur in hematopoietic cell lines： Raji cells and monocytic cell lines, which have average and above-average IFN-γ R2 surface expression, activate Statl similarly to HeLa cells and HeLa cells overexpressing IFNγR2, respectively. The rapid Statl activation seen in HeLa cells can be inhibited by overexpressing a chimeric IFN-γR2 chain that does not bind Jak2 or （when high concentrations of IFN-γ are used） by overexpressing IFN-γR1. These data are consistent with a model in which the recruitment of additional Jak2 activity to a signaling complex accelerates the rate of Statl activation. We conclude that the rate of activation of Statl in cells by IFN-γ can be modified by regulating either receptor chain and speculate that pharmacological agents which modify receptor chain expression may alter IFN-γ receptor signal transduction.

Assessment by meta-analysis of interferon-gamma for the diagnosis of tuberculous peritonitis

AIM:To investigate the performance and diagnostic accuracy of interferon-gamma(IFN-γ) for tuberculous peritonitis(TBP) by meta-analysis.METHODS:A systematic search of English language studies was performed.We searched the following electronic databases:MEDLINE,EMBASE,Web of Science,BIOSIS,LILACS and the Cochrane Library.The Standards for Reporting Diagnostic Accuracy initiative and Quality Assessment for Studies of Diagnostic Accuracy tool were used to assess the methodological quality of the studies.Sensitivity,specificity,and other measures of the accuracy of IFN-γ concentration in the diagnosis of peritoneal effusion were pooled using random-effects models.Receiver operating characteristic(ROC) curves were applied to summarize overall test performance.Two reviewers independently judged study eligibility while screening the citations.RESULTS:Six studies met the inclusion criteria.The average inter-rater agreement between the two reviewers for items in the quality checklist was 0.92.Analysis of IFN-γ level for TBP diagnosis yielded a summary estimate:sensitivity,0.93(95%CI,0.87-0.97);specificity,0.99(95%CI,0.97-1.00);positive likelihood ratio(PLR),41.49(95%CI,18.80-91.55);negative likelihood ratio(NLR),0.11(95%CI,0.06-0.19);and diagnostic odds ratio(DOR),678.02(95%CI,209.91-2190.09).χ 2 values of the sensitivity,specificity,PLR,NLR and DOR were 5.66(P = 0.3407),6.37(P = 0.2715),1.38(P = 0.9265),5.46(P = 0.3621) and 1.42(P = 0.9220),respectively.The summary receiver ROC curve was positioned near the desirable upper left corner and the maximum joint sensitivity and specificity was 0.97.The area under the curve was 0.99.The evaluation of publication bias was not significant(P = 0.922).CONCLUSION:IFN-γ may be a sensitive and specific marker for the accurate diagnosis of TBP.The level of IFN-γ may contribute to the accurate differentiation of tuberculosis(TB) ascites from non-TB ascites.

Effectiveness of interferon-gamma release assays for differentiating intestinal tuberculosis from Crohn's disease:A meta-analysis

AIM:To investigate the clinical usefulness of interferon-gamma release assays(IGRAs)in the differential diagnosis of intestinal tuberculosis(ITB)from Crohn’s disease(CD)by meta-analysis.METHODS:A systematic search of English language studies was performed.We searched the following databases:Medline,Embase,Web of Science and the Cochrane Library.The Standards for Reporting Diagnostic Accuracy initiative and Quality Assessment for Studies of Diagnostic Accuracy tool were used to assess the methodological quality of the studies.Sensitivity,specificity,and other measures of the accuracy of IGRAs in the differential diagnosis of ITB from CD were pooled and analyzed using random-effects models.Receiver operating characteristic curves were applied to summarize overall test performance.Two reviewers independently judged study eligibility while screening the citations.RESULTS:Five studies met the inclusion criteria.The average inter-rater agreement between the two reviewers for items in the quality checklist was 0.95.Analysis of IGRAs for the differential diagnosis of ITB from CD produced summary estimates as follows:sensitivity,0.74(95%CI:0.68-0.80);specificity,0.87(95%CI:0.82-0.90);positive likelihood ratio,5.98(95%CI:3.79-9.43);negative likelihood ratio,0.28(95%CI:0.18-0.43);and diagnostic odds ratio,26.21(95%CI:14.15-48.57).The area under the curve was 0.92.The evaluation of publication bias was not significant(P=0.235).CONCLUSION:Although IGRAs are not sensitive enough,they provide good specificity for the accurate diagnosis of ITB,which may be helpful in the differential diagnosis of ITB from CD.

The Expression of Interleukin-17, Interferon-gamma, and Macrophage Inflammatory Protein-3 Alpha mRNA in Patients with Psoriasis Vulgaris

Summary: To investigate the role of Interleukin-17 (IL-17), Interferon-gamma (IFN-γ), and macrophage inflammatory protein-3 alpha (MIP-3α) in the pathogenesis of psoriasis, reverse transcriptase-polymerase chain reaction (RT-PCR) was used to semi-quantitatively analyze the mRNA expression of IL-17, IFN-γ, and MIP-3α in 31 psoriatic lesions and 16 normal skin tissues. The results showed that the mRNA of the three cytokines was present in all specimens. And the expression level of IL-17 mRNA in skin lesions was 1.1416±0.0591, which was significantly higher than that in normal controls (0.8788±0.0344, P<0.001). The expression levels of IFN-γ mRNA were 1.1142±0.0561 and 0.9050±0.0263, respectively, with significant difference(P<0.001). And the expression levels of MIP-3α mRNA in psoriatic lesions was 1.1397±0.0521, which was markedly higher than that in normal controls (0.8681±0.0308, P<0.001). These findings indicate that up-regulated expression of IL-17, IFN-γ, and MIP-3α might be involved in the pathogenesis of psoriasis.

Retinal stem cells transplantation combined with copolymer-1 immunization reduces interferon-gamma levels in an experimental model of glaucoma

AIM: To explore the effect of immunization with copolymer-1 (COP-1) and retinal stem cells (RSCs) transplantation on interferon-gamma (IFN-gamma) levels in a rat experimental glaucoma model. METHODS: An experimental glaucoma was induced by argon laser photocoagulation of the episcleral veins and limbal plexus in the right eye of rats. Immediately following glaucoma induction, rats were immunized with COP-1. RSCs were cultured and transplanted intravitreally into the eyes of glaucoma model animals 1 week post-laser treatment. Six experimental groups were used: COP-1/RSC, PBS/RSC, COP-1/PBS, PBS/PBS, glaucoma model group, and a normal control group. The concentration of IFN-gamma in aqueous humor (AH) and serum was measured by enzyme-linked immunosorbent assay (ELISA) in each of the six groups. Retinal ganglion cell (RGC) survival was assessed by quantifying apoptosis using Hoechst staining. RESULTS: Concentrations of IFN-gamma in AH and serum of rats that had undergone glaucoma induction were higher than those of non-induced control rats. The concentrations of IFN-gamma in AH and serum of the COP-1/RSCs treated group were determined to be 2371.9ng/L and 710.9ng/L, respectively, which were significantly lower than those in the other treated groups (P<0.05). In fact, IFN-gamma levels in the dual treated group were reduced to background levels. The COP-1/RSC group had lower number of apoptotic RGCs than the other three experimental groups (P<0.05). CONCLUSION: The reduced levels of IFN-gamma in AH and serum of the COP-1/RSC group may be related to synergistic effects between RSCs transplantation and COP-1 immune modulation. It is likely that the lower levels of IFN-gamma prevented RGCs glaucomatous apoptasis.

Capillary Electrophoretic Immunoassay with Laser-induced Fluorescence Detection for Interferon-gamma

Capillary electrophoretic immunoassay with laser-induced fluorescence detection for recombinant human interferon-gamma (IFN-g) was established. The limits of detection for three forms of IFN-g are 6.9 ng/L, 5.7 ng/L and 5.0 ng/L, respectively.

Interferon-gamma release assays in tuberculous uveitis:a comprehensive review

·Tuberculous uveitis(TBU)comprises a broad clinical spectrum of ocular manifestations,making its diagnosis challenging.Ophthalmologists usually require evidence from investigations to confirm or support a clinical diagnosis of TBU.Since direct isolation of the causative organism from ocular specimens has limitations owing to the small volume of the ocular specimens,resultant test positivities are low in yield.Immunodiagnostic tests,including the tuberculin skin test and interferon-gamma release assays(IGRAs),can help support a clinical diagnosis of TBU.Unlike the tuberculin skin test,IGRAs are in vitro tests that require a single visit and are not affected by prior Bacillus Calmette-Guerin vaccination.Currently,available IGRAs consist of different techniques and interpretation methods.Moreover,newer generations have been developed to improve the sensitivity and ability to detect active tuberculosis.This narrative review collates salient practice points as a reference for general ophthalmologists,such as evidence for the utilization of IGRAs in patients with suspected TBU,and summarizes basic knowledge and details of clinical applications of these tests in a clinical setting.

Interferon-gamma and tumor necrosis factor-alpha synergistically enhance the immunosuppressive capacity of human umbilical-cordderived mesenchymal stem cells by increasing PD-L1 expression

BACKGROUND The immunosuppressive capacity of mesenchymal stem cells(MSCs)is dependent on the“license”of several proinflammatory factors to express immunosuppressive factors such as programmed cell death 1 ligand 1(PD-L1),which determines the clinical therapeutic efficacy of MSCs for inflammatory or immune diseases.In MSCs,interferon-gamma(IFN-γ)is a key inducer of PD-L1 expression,which is synergistically enhanced by tumor necrosis factor-alpha(TNF-α);however,the underlying mechanism is unclear.AIM To reveal the mechanism of pretreated MSCs express high PD-L1 and explore the application of pretreated MSCs in ulcerative colitis.METHODS We assessed PD-L1 expression in human umbilical-cord-derived MSCs(hUC-MSCs)induced by IFN-γand TNF-α,alone or in combination.Additionally,we performed signal pathway inhibitor experiments as well as RNA interference experiments to elucidate the molecular mechanism by which IFN-γalone or in combination with TNF-αinduces PD-L1 expression.Moreover,we used luciferase reporter gene experiments to verify the binding sites of the transcription factors of each signal transduction pathway to the targeted gene promoters.Finally,we evaluated the immunosuppressive capacity of hUC-MSCs treated with IFN-γand TNF-αin both an in vitro mixed lymphocyte culture assay,and in vivo in mice with dextran sulfate sodium-induced acute colitis.RESULTS Our results suggest that IFN-γinduction alone upregulates PD-L1 expression in hUC-MSCs while TNF-αalone does not,and that the co-induction of IFN-γand TNF-αpromotes higher expression of PD-L1.IFN-γinduces hUCMSCs to express PD-L1,in which IFN-γactivates the JAK/STAT1 signaling pathway,up-regulates the expression of the interferon regulatory factor 1(IRF1)transcription factor,promotes the binding of IRF1 and the PD-L1 gene promoter,and finally promotes PD-L1 mRNA.Although TNF-αalone did not induce PD-L1 expression in hUCMSCs,the addition of TNF-αsignificantly enhanced IFN-γ-induced JAK/STAT1/IRF1 activation.TNF-αupregulated IFN-γreceptor expression through activation of the nuclear factor kappa-B signaling pathway,which significantly enhanced IFN-γsignaling.Finally,co-induced hUC-MSCs have a stronger inhibitory effect on lymphocyte proliferation,and significantly ameliorate weight loss,mucosal damage,inflammatory cell infiltration,and up-regulation of inflammatory factors in colitis mice.CONCLUSION Overall,our results suggest that IFN-γand TNF-αenhance both the immunosuppressive ability of hUC-MSCs and their efficacy in ulcerative colitis by synergistically inducing high expression of PD-L1.

Influence of interferon-gamma on the differentiation of cholinergic neurons in rat embryonic basal forebrain and septal nuclei

BACKGROUND： Interferon-gamma （IFN-γ） can make neurons in basal forebrain and septal nuclei differentiate into cholinergic neurons by treating the cells in cerebral cortex of newborn rats, without the inhibition from IFN-γ antibody. The important effect of IFN-γ on the development and differentiation of neurons has been found by some scholars. OBJ EClIVE：To investigate whether IFN-γ has differentiational effect on cholinergic neurons in basal forebrain and septal nuclei, and make clear that the increased number of cholinergic neurons is resulted by cell differentiation or cell proliferation. DESIGN ： Controlled observation trial SETTING： Department of Cell Biology, Medical School, Beijing University MATERIALS： Sixty-eight female Wistar rats at embryonic 16 days, weighing 250 to 350 g, were enrolled in this study, and they were provided by the Experimental Animal Center, Medical School, Beijing University. IFN-γ was the product of Gibco Company. METHODS： This study was carried out in the Department of Cell Biology, Medical School, Beijing University and Daheng Image Company of Chinese Academy of Sciences during September 1995 to December 2002. The female Wistar rats at embryonic 16 days were sacrificed, and their fetuses were taken out. Primary culture of the isolated basal forebrain and septal nuclei was performed. The cultured nerve cells were assigned into 3 groups： control group （nothing added）, IFN-γ group（1×10^5 U/L interferon）, IFN-γ＋ IFN-γ antibody group （1 ×10^5 U/L IFN-γ ＋ IFN-γ antibody）. The specific marker enzyme （choline acetyl transferase） of cholinergic neuron was stained with immunohistochemical method. Choline acetyl transferase positive cells were counted, and ^14C-acetyl CoA was used as substrate to detect the activity of choline acetyl transferase, so as to reflect the differentiational effect of IFN-γ on cholinergic neuron in basal forebrain and septal nuclei. Flow cytometry was used to analyze cell circle and detect the proliferation of nerve cells. Nerve cells were marked with MAP2 and counted to evaluate the neuronal proliferation in basal forebrain and septal nuclei. MAIN OUTCOME MEASURES： Effect of interferon-γ on the number and activity of choline acetyl transferase-positive ceils in basal forebrain and septal nuclei as well as the effect on neuronal proliferation. RESULTS ： ① Nerve cells in the basal forebrain and septal nuclei of IFN-γ group grew well compared with control group.②The differentiation of cholinergic neurons： The number and activity of choline acetyl transferase positive cells in IFN-γ group were significantly higher than those in the control group [（49.30 ±4.92） /100 cells vs （7.50±1.58） /100 cells; （2 049.00±12.30） min^-1 vs （1 227.30±12.59） min^-1, p 〈 0.01], while there was no significant difference in the number and activity of choline acetyl transferase positive cells between IFN-γ ＋ IFN-γ antibody group and control group（P 〉 0.05）. ③The proliferation of cholinergic neurons： Cell percentage was 17.2% and 19.8% at S-stage, 6.2% and 6.1% at G2＋M stage in the control group and IFN-γ group respectively, without significant difference （P 〉 0.05）. CONCLUSION ： IFN-γ does not promote the neuronal proliferation in basal forebrain and septal nuclei, and the increased expression of cholinergic neurons is not resulted by the increase in the number of neurons, but its differentiation.

Profiles of interferon-gamma and interleukin-2 in patients after allogeneic hematopoietic stem cell transplantation

BACKGROUND Allogeneic hematopoietic stem cell transplantation(allo-HSCT)may be related to the occurrence of complications,including graft-versus-host disease(GvHD)and infections.The pathogenesis of acute GvHD is connected with T lymphocytes,which identify alloantigens on host's antigen-presenting cells,activate production of interferon-gamma(IFN-gamma)and interleukin-2(IL-2),and act on the immune effector cells and damage tissues and organs.AIM The aim of the study was to investigate and distinguish serum concentration profiles of IFN-gamma and IL-2 within a 30-d period after allo-HSCT.METHODS We enrolled 62 patients,i.e.,30(48%)male and 32(52%)female subjects[median age 49.5(19-68)years],after allo-HSCT from siblings(n=12)or unrelated donors(n=50)due to acute myeloid leukemia with myeloablative conditioning(n=26;42%)and with non-myeloablative conditioning(n=36;58%).All patients were given standard immunosuppressive therapy with cyclosporin-A and methotrexate and pre-transplant antithymocyte globulin in the unrelated setting.Blood samples were collected pre-transplant before and after(on day-1)the conditioning therapy and on days+2,+4,+6,+10,+20,and+30 after allo-HSCT.Serum levels of IL-2 and IFNgamma were determined using ELISA.RESULTS Patients were divided into four groups depending on the presence of acute GvHD and clinical manifestations of infection.Group I included patients with neither acute GvHD nor infections[n=15(24%)],group II consisted of patients with infections without acute GvHD[n=17(27%)],group III was comprised of patients with acute GvHD without infections[n=9(15%)],and group IV included patients with both acute GvHD and infections[n=21(34%)].IFN-gamma concentrations were higher in Group II than in other groups on days+20(P=0.014)and+30(P=0.008).Post-hoc tests showed lower concentrations of IFN-gamma on day+30 in groups I(P=0.039)and IV(P=0.017)compared to group II.The levels of IL-2 were mostly undetectable.CONCLUSION Serum levels of IFN-gamma following allo-HSCT progressively escalate.High serum levels of IFN-gamma are related to infectious complications rather than acute GvHD.Serum concentrations of IL-2 in most patients are undetectable.

Interferon-Gamma Release Assay is Not Appropriate for the Diagnosis of Active Tuberculosis in High-Burden Tuberculosis Settings： A Retrospective Multicenter Investigation

Background：Interferon-gamma release assay （IGRA） has been used in latent tuberculosis （TB） infection and TB diagnosis,but the results from different high TB-endemic countries are different.The aim of this study was to investigate the value of IGRA in the diagnosis of active pulmonary TB （PTB） in China.Methods：We conducted a large-scale retrospective multicenter investigation to further evaluate the role of IGRA in the diagnosis of active PTB in high TB-epidemic populations and the factors affecting the performance of the assay.All patients who underwent valid T-SPOT.TB assays from December 2012 to November 2015 in six large-scale specialized TB hospitals in China and met the study criteria were retrospectively evaluated.Patients were divided into three groups：Group 1,sputum culture-positive PTB patients,confirmed by positive Mycobacterium tuberculosis sputum culture;Group 2,sputum culture-negative PTB patients;and Group 3,non-TB respiratory diseases.The medical records of all patients were collected.Chi-square tests and Fisher＇s exact test were used to compare categorical data.Multivariable logistic analyses were performed to evaluate the relationship between the results of T-SPOT in TB patients and other factors.Results：A total of 3082 patients for whom complete information was available were included in the investigation,including 905 sputum culture-positive PTB cases,914 sputum cultmre-negative PTB cases,and 1263 non-TB respiratory disease cases.The positive rate of T-SPOT.TB was 93.3% in the culture-positive PTB group and 86.1% in the culture-negative PTB group.In the non-PTB group,the positive rate of T-SPOT.TB was 43.6%.The positive rate of T-SPOT.TB in the culture-positive PTB group was significantly higher than that in the culture-negative PTB group （x2 =25.118,P 〈 0.01）,which in turn was significantly higher than that in the non-TB group （x2 =566.l 16,P 〈 0.01）.The overall results were as follows：sensitivity,89.7%;specificity,56.37%;positive predictive value,74.75%;negative predictive value,79.11%;and accuracy,76.02%.Conclusions：High false-positive rates of T-SPOT.TB assays in the non-TB group limit the usefulness as a single test to diagnose active TB in China.We highly recommend that IGRAs not be used for the diagnosis of active TB in high-burden TB settings.

Interferon-gamma inhibits aldehyde dehydrogenase^(bright)cancer stem cells in the 4T1 mouse model of breast cancer

Background:Despite improvements in disease diagnosis,treatment,and prognosis,breast cancer is still a leading cause of cancer death for women.Compelling evidence suggests that targeting cancer stem cells(CSCs)have a crucial impact on overcoming the current shortcomings of chemotherapy and radiotherapy.In the present study,we aimed to study the effects of T cells and a critical anti-tumor cytokine,interferon-gamma(IFN-γ),on breast cancer stem cells.Methods:BALB/c mice and BALB/c nude mice were subcutaneously injected with 4T1 tumor cells.Tumor growth and pulmonary metastasis were assessed.ALDEFLOUR™assays were performed to identify aldehyde dehydrogenase^(bright)(ALDH^(br))tumor cells.ALDH^(br)cells as well as T cells from tumor-bearing BALB/c mice were analyzed using flow cytometry.The effects of CD8^(+)T cells on ALDH^(br)tumor cells were assessed in vitro and in vivo.The expression profiles of ALDH^(br)and ALDH^(dim)4T1 tumor cells were determined.The levels of plasma IFN-γwere measured by enzyme-linked immunosorbent assay,and their associations with the percentages of ALDH^(br)tumor cells were evaluated.The effects of IFN-γon ALDH expression and the malignancy of 4T1 tumor cells were analyzed in vitro.Results:There were fewer metastatic nodules in tumor-bearing BALB/c mice than those in tumor-bearing BALB/c nude mice(25.40 vs.54.67,P<0.050).CD8^(+)T cells decreased the percentages of ALDH^(br)4T1 tumor cells in vitro(control vs.effector to target ratio of 1:1,10.15%vs.5.76%,P<0.050)and in vivo(control vs.CD8^(+)T cell depletion,10.15%vs.21.75%,P<0.001).The functions of upregulated genes in ALDH^(br)4T1 tumor cells were enriched in the pathway of response to IFN-γ.The levels of plasma IFN-γdecreased gradually in tumor-bearing BALB/c mice,while the percentages of ALDH^(br)tumor cells in primary tumors increased.IFN-γat a concentration of 26.68 ng/mL decreased the percentages of ALDH^(br)4T1 tumor cells(22.88%vs.9.88%,P<0.050)and the protein levels of aldehyde dehydrogenase 1 family member A1 in 4T1 tumor cells(0.86 vs.0.49,P<0.050)and inhibited the abilities of sphere formation(sphere diameter<200μm,159.50 vs.72.0;≥200μm,127.0 vs.59.0;both P<0.050)and invasion(89.67 vs.67.67,P<0.001)of 4T1 tumor cells.Conclusion:CD8^(+)T cells and IFN-γdecreased CSC numbers in a 4T1 mouse model of breast cancer.The application of IFN-γmay be a potential strategy for reducing CSCs in breast cancer.

慢加急性乙型肝炎肝衰竭患者细菌感染病原菌分布及血清PCT、IFN-γ和IL-6水平变化分析

目的调查慢加急性乙型肝炎肝衰竭(HBV-ACLF)患者细菌感染病原菌分布特征,分析血清降钙素原(PCT)、干扰素-γ(IFN-γ)和白细胞介素-6(IL-6)水平预测细菌感染的效能。方法2019年12月~2023年1月我院收治的HBV-ACLF患者86例,常规分离和鉴定菌种,采用电化学发光免疫法检测血清PCT水平,采用ELISA法检测血清IFN-γ和IL-6水平,应用受试者工作特征(ROC)曲线评估血清指标联合预测HBV-ACLF患者发生细菌感染的效能。结果本组发生细菌感染者37例,未发生明确感染者49例;在37例HBV-ACLF并发细菌感染患者中,共检出68株感染病原菌,其中革兰氏阴性菌41株(60.3%),革兰氏阳性菌27株(39.7%);感染组血清PCT、IFN-γ和IL-6水平分别为(10.9±3.1)μg/L、(46.5±1.9)pg/mL和(16.9±1.6)pg/mL,均显著高于未感染组【分别为(0.9±0.1)μg/L、(20.1±2.4)pg/mL和(4.8±0.9)pg/mL,P<0.05】,28 d和90 d病死率分别为67.6%和75.7%,均显著高于未感染组(分别为8.2%和12.2%,P<0.05);经ROC分析显示,分别以PCT>3.3μg/L、IFN-γ>45.5 pg/mL和IL-6>15.4pg/mL为截断点,其联合预测HBV-ACLF患者并发细菌感染的曲线下面积(AUC)为0.874,诊断的灵敏度为93.6%,特异度为84.1%。结论HBV-ACLF患者细菌感染的病原菌分布具有一定的特征性,以革兰氏阴性菌感染为主,除检测血清PCT水平外,监测血清IFN-γ和IL-6水平可能有助于早期发现细菌感染而给予预防性处理,或能提供生存率。

TB-DNA、IGRAs、TB-Ab在肺结核中的诊断价值

目的探究结核分枝杆菌核酸(TB-DNA)、γ-干扰素释放试验(IGRAs)和结核分歧杆菌抗体(TB-Ab)检测在肺结核中的诊断价值。方法收集2022年6月至2023年1月莆田学院附属医院收治的154例疑似肺结核患者,所有患者均接受TB-DNA、IGRAs和TB-Ab检测,评价不同检测方法的检测阳性率、诊断肺结核的灵敏度、特异度、阳性预测值、阴性预测值及诊断准确率;采用受试者工作特征曲线(ROC)评估不同检测方法对肺结核的诊断效能。结果154例疑似肺结核患者中,107例确诊为肺结核(肺结核组),47例确诊为非肺结核的其他肺部疾病(对照组)。肺结核组TB-DNA、IGRAs和TB-Ab阳性率均高于对照组,差异有统计学意义(P<0.05)。TB-DNA、IGRAs检测诊断肺结核的灵敏度和诊断准确率均高于TB-Ab检测,差异有统计学意义(P<0.05),TB-DNA检测诊断肺结核的阴性预测值高于TB-Ab检测,差异有统计学意义(P<0.05)。ROC曲线分析结果显示,TB-DNA诊断肺结核的AUC为0.851(明显大于IGRAs诊断的0.770)、TB-Ab诊断的0.770,IGRAs诊断的AUC明显大于TB-Ab,差异均有统计学意义(P<0.05)。IGRAs+TB-DNA联合诊断的灵敏度、诊断准确率明显高于IGRAs单独检测(P<0.05);IGRAs+TB-Ab联合诊断的灵敏度明显高于IGRAs、TB-Ab单独检测,诊断准确率明显高于TB-Ab单独检测(P<0.05);TB-DNA+TB-Ab联合诊断的灵敏度、阴性预测值、诊断准确率明显高于TB-Ab单独检测(P<0.05)。TB-DNA、IGRAs、TB-Ab两两联合检测和三者联合检测诊断肺结核的灵敏度、特异度、阳性预测值、阴性预测值及诊断准确率比较,差异均无统计学意义(P>0.05)。结论相比TB-Ab和IGRAs检测,TB-DNA检测对肺结核的诊断价值更高;TB-DNA、IGRAs、TB-Ab两两联合检测对肺结核的诊断效能高于单一指标检测。

消瘿散结汤联合左甲状腺素钠治疗桥本氏甲状腺炎合并甲状腺功能减退床疗效分析

目的探讨消瘿散结汤联合左甲状腺素钠片治疗桥本氏甲状腺炎(hashimoto's thyroiditis,HT)合并甲状腺功能减退患者的临床疗效。方法选择2022年1月—2022年12月辽宁中医药大学附属第二医院收治的80例HT合并甲状腺功能减退患者,按照随机数字表法为对照组和观察组各40例。对照组予以左甲状腺素钠治疗;观察组在对照组基础上加用消瘿散结汤口服治疗。观察两组临床疗效、中医证候积分变、血清游离三碘甲腺原氨酸(FT3)、游离甲状腺素(FT4)、促甲状腺激素(TSH)、抗甲状腺球蛋白抗体(TGAb)、抗甲状腺过氧化物酶抗体(TPOAb)、血清γ干扰素(IFN-γ)、白细胞介素-2(IL-2)、白细胞介素-4(IL-4)、白细胞介素-10(IL-10)等。结果治疗后,观察组总有效率高于对照组(P<0.05)。两组TGAb、TPOAb抗体滴度均降低,观察组低于对照组(P<0.05);两组中医证候总积分均降低,观察组低于对照组(P<0.05);两组IFN-γ、IL-2水平均降低(P<0.05),观察组低于对照组(P<0.05)。结论消瘿散结汤联合左甲状腺素钠片治疗HT合并甲状腺功能减退有效,且可在一定程度上降低TGAb、TPOAb及IL-2、IL-4、IL-10水平,改善甲状腺功能及临床症状,提升临床治疗有效率,值得推广。

藏獒IFN-γ基因在毕赤酵母中的表达与抗病毒活性分析

采用毕赤酵母表达系统表达藏獒γ-干扰素(Tibetan mastiff interferon gamma,TmIFN-γ)并分析其生物活性,为抗病毒生物制品的研发与临床应用奠定基础。将构建的TmIFN-γ的真核表达质粒pPIC9k-TmIFN-γ转化至毕赤酵母GS115细胞,用不同浓度的G418筛选得到多拷贝重组菌株;利用甲醇进行诱导表达,以镍层析柱纯化目的蛋白,对表达产物进行SDS-PAGE和Western blotting鉴定;通过CCK-8试验检测重组TmIFN-γ蛋白的细胞毒性作用,利用VSV-MDBK系统检测其抗病毒生物活性。结果显示,成功筛选得到高拷贝pPIC9k-TmIFN-γ重组转化毕赤酵母菌株,利用甲醇诱导可获得TmIFN-γ重组蛋白,该蛋白对细胞无毒性,具有显著抗病毒活性,其抗病毒生物效价为1.727×10~5IU/mL。本研究获得了具有抗病毒活性的TmIFN-γ,有助于进一步探索TmIFN-γ的功能和开发新型基因工程抗病毒药物。

肺结核患者血清IP-10、SOCS1及CA125表达意义及其与引发支气管狭窄的相关性分析

目的分析肺结核患者血清γ干扰素诱导蛋白10(IP-10)、细胞因子信号转导抑制物1(SOCS1)及糖类抗原125(CA125)表达意义及其与引发支气管狭窄的相关性。方法回顾性选择2021年1月至2023年1月河北省胸科医院收治的116例肺结核患者作为观察组,同期116名健康体检者作为对照组。根据观察组患者胸部影像学检查结果,分为重度组(n=26)和轻中度组(n=90);通过纤维支气管镜检查及活检,判断患者是否存在支气管狭窄,分为支气管狭窄组(n=42)与单纯肺结核组(n=74)。检测所有入选者血清IP-10、SOCS1及CA125水平,比较血清IP-10、SOCS1及CA125在对照组与观察组、不同严重程度的肺结核患者中的差异性;比较支气管狭窄组与单纯肺结核组第1秒用力呼气量(FEV1)、用力肺活量(FVC)、最大呼气峰流速(PEF);分析血清IP-10、SOCS1及CA125与肺结核患者肺功能相关指标及其并发支气管狭窄的相关性。采用受试者工作特征(ROC)曲线分析血清IP-10、SOCS1及CA125水平预测肺结核患者发生支气管狭窄的效能。结果观察组血清IP-10、CA125水平分别为(95.84±12.58)ng/L、(1.89±0.86)U/mL,均高于对照组[(71.25±5.63)ng/L、(0.74±0.23)U/mL],SOCS1水平为(165.32±7.95)μg/L,低于对照组[(252.54±24.71)μg/L],差异均有统计学意义(P<0.05)。重度组肺结核患者血清IP-10、CA125水平分别为(106.74±15.01)ng/L、(2.67±1.12)U/mL,均高于轻中度组[(89.72±8.16)ng/L、(1.45±0.60)U/mL],SOCS1水平为(154.12±6.07)μg/L,低于轻中度组[(200.75±15.83)μg/L],差异均有统计学意义(P<0.05)。支气管狭窄组FEV1、FVC、PEF均小于单纯肺结核组,差异均有统计学意义(P<0.05)。经Pearson相关性分析,肺结核患者FEV1、FVC、PEF与血清IP-10、CA125水平呈正相关(P<0.05),与SOCS1水平呈负相关(P<0.05)。经ROC曲线分析,血清IP-10、SOCS1联合CA125预测肺结核患者发生支气管狭窄的敏感度为89.12%,特异度为48.63%,曲线下面积为0.924。结论肺结核患者血清IP-10、CA125水平明显升高,SOCS1水平降低,与病情严重程度相关,其联合预测支气管狭窄具有较好的效能。