Performance and correlation of interferon gamma release assays and tuberculin skin test in HIV-infected children and adolescents with immune reconstitution

Objective:To evaluate the performance of interferon gamma release assays and tuberculin skin test in HIV-infected children and adolescents with immune reconstitution.Methods:A cross-sectional study was conducted in HIV-infected patients aged 5-18 years receiving antiretroviral treatment with CD4 T-lymphocytes>25%or>500 cells/mm3 for at least 6 months.QuantiF ERON-TB Gold,T-SPOT.TB,and tuberculin skin test were performed in each patient.Results:A total of 50 patients were enrolled with median age of 13.7 years,CD4 counts of 753(IQR:587-989)cells/mm3.Among 27 patients with tuberculosis(16)or tuberculosis exposure(11),8(29.6%)were positive to at least one test,2(7.4%)were positive QuantiFERON-TB Gold,3(11.1%)positive T-SPOT.TB,and 7(25.9%)had tuberculin skin test≥5 mm.Among 23 patients without history of tuberculosis or exposure,all had negative interferon gamma release assays,while 2(8.7%)had positive tuberculin skin test.Conclusions:All tests had low sensitivity despite immune reconstitution.

Effect of recombinant human growth hormone and interferon gamma on hepatic collagen synthesis and proliferation of hepatic stellate cells in cirrhotic rats

BACKGROUND: Fibrosis plays a key role in the development of liver cirrhosis. In this study, we investigated the effect of growth hormone and interferon gamma on hepatic collagen synthesis and the proliferation of hepatic stellate cells in a cirrhotic rat model. METHODS: Cirrhosis was induced in rats using carbon tetrachloride. Rats were simultaneously treated with daily subcutaneous injections of recombinant human growth hormone or interferon gamma combined with recombinant human growth hormone. The control group was given saline. The relative content of type I and type IV collagen was assessed by indirect immunofluorescence analysis. Activated hepatic stellate cells were prepared from cirrhotic rats. The 3-(4, 5-dimethyl-2-thiazolyl)-2, 5-diphenyl-2H-tetrazolium bromide (MTT) method was used to assess the effects of recombinant human growth hormone and interferon gamma on these cells in vitro. RESULTS: Both qualitative and quantitative analysis showed that type I and type IV collagen secretion increased with time after recombinant human growth hormone administration and was significantly higher than control and recombinant human growth hormone combined with interferon gamma administration. In vitro, recombinant human growth hormone significantly stimulated hepatic stellate cell proliferation in a concentration-dependent manner (10 -3 -10 -1 mg/100 μL), andinterferon gamma (10 -2 -10 -1 μg/100 μL) significantly inhibited their growth compared to the control group. Interferon gamma combined with recombinant human growth hormone eliminated this growth-promoting effect to a certain degree in a concentration-dependent manner (10 -1 μg/100 μL, P<0.05, 10 -2 -10 -3 μg/100 μL, P>0.05) and a time-dependent manner (P<0.05). CONCLUSIONS: Recombinant human growth hormone increased collagen secretion in cirrhotic rats in vivo and promoted the proliferation of hepatic stellate cells from cirrhotic rats in vitro. It is possible that concurrent interferon gamma therapy can offset these side-effects of recombinant human growth hormone.

Clinical and bacteriological correlates of whole blood interferon gamma(IFN-γ) in newly detected cases of pulmonary TB

Objective:To determine the relationship of the capacity to produce interferon gamma(IFN-γ) in whole blood,bacteriological,hematological,radiographic and clinical presentations in new,HIV seronegative cases of pulmonary tuberculosis(TB).Methods:80 cases and 50 control subjects aged 15 years onwards,representative of Kasturba Hospital and Nursing schools of Wardha district of Maharashtra state in India were examined for their health condition with standard methodology.Results:Among these TB patients,73.8%were Quantiferon-TB gold (QFT) positive with IFN-γconcentration as 0.35 IU or more and there was none in healthy controls.The mean IFN-γconcentrations varied between 9.58 IU(50-59 yrs) and 2.58 IU(≥60 yrs),showing no trend.The differences in positivity and mean IFN-γconcentrations were statistically insignificant.Both the QFT positivity and IFN-γconcentrations were higher in normal lymphocyte percent as compared to below and above normal,but differences were not statistically significant.Conclusions:The IFN-γconcentrations are not correlated with any of the predictors of disease severity studied,the levels are significantly higher in observation group as compared to healthy group.

Inhibitory effect of gold nanoparticles conjugated with interferon gamma and methionine on breast cancer cell line

Objective: To develop a gold nanoparticles complex conjugated with interferon-gamma(IFN-g) and methionine along with application of hyperthermia using near-infrared laser beams for the treatment of cancer cells.Methods: Gold nanorods(10 nm) were conjugated with IFN-g and methionine using carbodiimide family and characterized after purification by dialysis bags. Breast cancer cells were cultured and incubated with gold nanorods at different concentrations followed by irradiation with near-infrared laser beam. Samples were then evaluated for their viability in order to determine the effect of treatment and variables by MTT assy.Results: Zetasizer results confirmed the conjugation of gold nanorods with methionine and IFN-g. The median percentage of cell viability in 0.30 mg/m L concentration of gold nanorods was 82%. The cell viability reached to 85% at the same concentration of gold nanorods, which existed in the assayed complex. The results of MTT assay showed that the 0.60 mg/m L concentration of gold nanoparticles complex was toxic on tumor cells(P < 0.05). After exposure to hyperthermia, the viability of cells at 6 min decreased to77% in 0.30 mg/m L concentration of gold nanorods complex.Conclusions: The size and concentration of gold nanorods was not cytotoxic. However,their presence during irradiation near-infrared laser increased the number of dead cells during the treatment of cells.

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.

Association between interferon gamma receptor 1-56C/T gene polymorphism and tuberculosis susceptibility： a meta-analysis

Background Genetic variations in the interferon-gamma （IFN-γ） receptor 1 gene （IFNGR1） may contribute to tuberculosis （TB） risk in different populations.Many studies have investigated the relationship between IFNGR1 56C/T polymorphism and the susceptibility to TB,but have yielded conflicting results.A comprehensive meta-analysis is needed to provide a more accurate estimation of the relationship between them.Methods A literature search based on a combination of manual and computer-based methods was conducted on four English databases （PubMed,Science Direct,SpringerLink,and EBSCO） and three Chinese databases （Wanfang,CQVIP,and Chinese National Knowledge Infrastructure databases）.Pooled odds ratios （ORs） and 95％ confidence intervals （95％ Cls） were calculated using either the fixed-effects model or the random-effects model for different genetic models based on the heterogeneity examination.Results A total of six studies comprising 1 497 confirmed TB cases and 1 802 controls were included in this meta-analysis.Overall,no significant association was observed between IFNGR1-56C/T polymorphism and TB susceptibility （C vs.T,OR=0.90,95％ Cl 0.69-1.17; CC vs.TT,OR=0.87,95％ Cl 0.65-1.18; TC vs.TT,OR=-1.031,95％ Cl 0.872-1.219; CC＋TC vs.TT,OR=0.89,95％ Cl 0.64-1.26; CC vs.TC＋TT,OR=0.92,95％ Cl 0.66-1.29）.In subgroup analysis,a significant association was found in the dominant model （CC＋TC vs.TT,OR=1.24,95％ Cl 1.02-1.51） in Africans,but not in Asians or Caucasians.Conclusions Our meta-analysis did not provide enough powerful evidence to identify a significant association between IFNGR1-56C/T polymorphism and TB susceptibility in the overall population.In subgroup analysis,it indicates that IFNGR1-56C/T is possibly associated with increased TB risk in Africans,but not in Asians or Caucasians.However,larger sample size and better-designed case-control studies are needed to validate these findings.

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.

Animal experiment and clinical study of effect of gamma-interferon on hepatic fibrosis

AIM: To evaluate the antifibrotic effect of different doses of recombinant human Gamma-Interferon (IFN-gamma) in two rat models of hepatic fibrosis, and to observe its effect on moderate chronic hepatitis B virus fibrosis. METHODS: Hepatic fibrosis was successfully induced in 150 and 196 rats by subcutaneous injection of carbon tetrachloride (CCl4) and intraperitoneal injection of dimethylnitrosamine (DMN), respectively. Each of the two model groups was divided into: (1) fibrotic model group; (2) colchicine treatment group (0.1 mg/kg/day, gastrogavage for 8 weeks); (3) high-dose IFN-gamma group (15 MU/kg per day, i.m. for 8 weeks); (4) medium-dose IFN-gamma group (5 MU/kg daily, i.m. for 8 weeks); and (5) Y low-dose IFN-gamma group (1.67 MU/kg daily, i.m. for 8 weeks). Another group of 10 rats without any treatment was used as normal controls. At the end of the experiment, semi-quantitative histopathological scores of inflammation and fibrosis, liver alpha smooth muscle actin (alpha-SMA) expression level, liver hydroxyl proline content and serum hyaluronic acid levels were compared. And 47 medium chronic hepatitis B viral fibrosis patients were studied. They were given IFN-gamma treatment, 100 MU/day i.m. for the first three months and 100 MU qod i.m. for the next six months. Semi-quantitative pathological scores of inflammation and fibrosis and serum hepatic fibrosis indices were compared within the 9 months. RESULTS: In animal experiment, the pathological fibrosis scores and liver hydroxyl proline content were found to be significantly lower in rats treated with different doses of IFN-gamma as compared with rats in fibrotic model group induced by either CCl4 or DMN, in a dose-dependent manner. For CCl4-induced model, pathological fibrosis scores in high, medium and low doses IFN-gamma groups were 5.10 +/- 2.88, 7.70 +/- 3.53 and 8.00 +/- 3.30, respectively, but the score was 14.60 +/- 7.82 in fibrotic model group. Hydroxyl proline contents were 2.83 +/- 1.18, 3.59 +/- 1.22 and 4.80 +/- 1.62, in the three IFN-gamma groups, and 10.01 +/- 3.23 in fibrotic model group. The difference was statistically significant (P【0.01). Similar results were found in DMN-induced model. Pathological fibrosis scores were 6.30 +/- 0.48, 8.10 +/- 2.72 and 8.30 +/- 2.58, in high, medium and low doses IFN-gamma groups, and 12.60 +/- 3.57 in fibrotic model group. Hydroxyl proline contents were 2.72 +/- 0.58, 3.14 +/- 0.71 and 3.62 +/- 1.02, in the three IFN-gamma groups, and 12.79 +/- 1.54 in fibrotic model group. The difference was statistically significant (P【0.01).Serum hepatic fibrosis indices decreased significantly in the 47 patients after IFN-gamma treatment (HA: 433.38 +/- 373.00 vs 281.57 +/- 220.48; LN: 161.22 +/- 41.02 vs 146 +/- 35 +/- 44. 67; PC III: 192.59 +/- 89.95 vs 156.98 +/- 49.22; C-I: 156.30 +/- 44.01 vs 139.14 +/- 34.47) and the differences between the four indices were significant (P 【0.05). Thirty-three patients received two liver biopsies, one before and one after IFN-gamma treatment. In thirty of 33 patients IFN-gamma had better effects according to semi-quantitative pathological scores (8.40 +/- 5.83 vs 5.30 +/- 4.05, P【0.05). CONCLUSION: All the three doses of IFN-gamma are effective in treating rat liver fibrosis induced by either CCl4 or DMN, the higher the dose, the better the effect. And IFN-gamma is effective for patients with moderate chronic hepatitis B viral fibrosis.

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.

Effect of neuropeptide Y on white matter demyelination and serum interleukin-4 and gamma-interferon levels in the guinea pig with experimental allergic encephalomyelitis

BACKGROUND： Neuropeptide Y （NPY） may influence differentiation of Th cells immunological pathology of experimental allergic encephalomyelitis （EAE） is differentiation of Th cells It is assumed that the related to abnormal OBJECTIVE： To investigate the effect of NPY on white matter demyelination, the serum levels interleukin-4 （IL-4） and gamma-interferon （IFN-γ ）, as well as EAE pathogenesis in an EAE guinea pig model following NPY injection into the lateral cerebral ventricle. DESIGN, TIME AND SETTING： A randomized controlled animal study, which was performed in the Infection Immunity Animal Laboratory, Affiliated Hospital of Luzhou Medical College, China, from October 2005 to April 2006. MATERIALS： Thirty healthy female guinea pigs of 8-12 weeks of age, and 10 healthy female rats of three months of age were used. NPY was provided by Sigma Company, USA. NPY kit was provided by Beijing Huaying Biotechnology Institute, China. METHODS： Thirty guinea pigs were randomly divided into three groups： normal control group, EAE model group, and NPY intervention group （n =10 per group）. Normal control group and EAE model group： Saline （10μ L, once） was injected into the lateral cerebral ventricle. After one week, the same volume of Freund＇s adjuvant complete was either injected subcutaneously into two post-palms or EAE was modeled. NPY intervention group： EAE was modeled after one week and NPY was injected （10 μ L of 6 nmol NPY, once） into the lateral cerebral ventricle. Myelin basic protein （MBP） antigen made from rat spinal cord homogenate and Freund＇s adjuvant complete were injected subcutaneously into both post-palms （0.2 mL per palm） to establish the EAE model. MAIN OUTCOME MEASURES： White matter demyelination of the cerebrum, cerebellum, brain stem, and spinal cord were observed by light microscopy after HE staining. Levels of serum IFN-γ and IL-4 were detected by the double antibody sandwich ABC-ELISA technique. NPY content was detected by radioimmunoassay. RESULTS： Pathological alterations in the NPY intervention groups were reduced compared to those in the EAE model group, suggesting a reduction and remission of white matter demyelination with NPY treatment. When compared to the model group, the serum IL-4 level was increased in the NPY intervention group during the high-frequent EAE stage （P 〈 0.01）, but the serum IFN-γ level was decreased （P 〈 0.01）. Furthermore, the EAE latency was prolonged （P 〈 0.01）, the neurological scores were decreased in the high-frequent EAE stage （P 〈 0.01）, and the death rate was decreased （P 〈 0.05）. NPY content and the serum IL-4 level at the peak stage were positively correlated with those in the latent phase （r =0.863-0.900, P 〈 0.01）, but negatively correlated with neurological scores at the peak stage （r=- -0.068 to -0.863, P 〈 0.05-0.01）. The IFN-γ level at the peak stage was negatively correlated to that in the latent phase （r = -0.683-0.650, P 〈 0.05）, but positively correlated to neurological scores at the peak stage （r =0.975, 0.845, P 〈 0.05）. CONCLUSION： NPY injection into the lateral cerebral ventricle can promote the secretion of IL-4, inhibit the production of IFN-γ, relieve white matter demyelination, and inhibit EAE attack in an experimental model of EAE.

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。

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。

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。

Low interleukin-10 level indicates a good prognosis in Salmonella enterica serovar typhimurium-induced pediatric hemophagocytic lymphohistiocytosis:A case report

BACKGROUND Secondary hemophagocytic lymphohistiocytosis(sHLH)triggered by Salmonella enterica serovar Typhimurium is rare in pediatric patients.There is no consensus on how to treat S.typhimurium-triggered sHLH.CASE SUMMARY A 9-year-old boy with intermittent fever for 3 d presented to our hospital with positive results for S.typhimurium,human rhinovirus,and Mycoplasma pneumoniae infections.At the time of admission to our institution,the patient’s T helper 1/T helper 2 cytokine levels were 326 pg/mL for interleukin 6(IL-6),9.1 pg/mL for IL-10,and 246.7 pg/mL for interferon-gamma(IFN-γ),for which the ratio of IL-10 to IFN-γwas 0.04.In this study,the patient received meropenem,linezolid,and cefoperazone/sulbactam in combination with high-dose methylprednisolone therapy(10 mg/kg/d for 3 d)and antishock supportive treatment twice.After careful evaluation,this patient did not receive HLH chemotherapy and recovered well.CONCLUSION S.Typhimurium infection-triggered sHLH patient had a ratio of IL-10 to IFN-γ≤1.33,an IL-10 concentration≤10.0 pg/mL,and/or an IFN-γconcentration≤225 pg/mL at admission.Early antimicrobial and supportive treatment was sufficient,and the HLH-94/2004 protocol was not necessary under these conditions.

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。

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。