ABSTRACT
Objective To analyze the value of peripheral blood T-lymphocyte subsets and cytokines in the diagnosis of Mycoplasma pneumoniae pneumonia(MPP)in children.Methods A total of 120 children with MPP who were admitted to the hospital from January 2020 to January 2023 were selected as the observation group,80 children with pulmonary tuberculosis(TB)were selected as the control group.During the same pe-riod,120 healthy children who were examined at the hospital check-up center were selected as the health group.The clinical data from each group were retrospective analyzed,and fasting venous blood from subjects was collected.The levels of T-lymphocyte subsets CD3+,CD4+and CD8+were detected by flow cytometry in each group,and the CD4+/CD8+was calculated.The levels of interferon(IFN)-y,interleukin-8(IL-8),inter-leukin-10(IL-10)and interleukin-13(IL-13)were measured by enzyme-linked immunosorbent assay,and the levels of peripheral blood T-lymphocyte subsets and cytokines in each group were compared.The receiver op-erating characteristic(ROC)curve was used to assess the value of peripheral blood T-lymphocyte subsets and cytokines in the diagnosis of MPP in children.Results The levels of CD3+,CD4+,CD4+/CD8+,IL-13,and IL-10 in the observation group and control group were significantly lower than those in the health group(P<0.05),while CD8+,IL-8,and IFN-γ levels were significantly higher than those in the health group(P<0.05).Compared with the control group,the observation group had higher levels of CD3+,CD8+,IL-8,IFN-γand lower levels of CD4+,CD4+/CD8+(P<0.05),and there were no statistically significant differences in IL-8 and IL-13 levels between the two groups(P>0.05).The ROC curve results showed that the area under the curve(AUC)or CD3+,CD4+,CD8+,CD4+/CD8+,IL-8,IFN-γ,IL-13,and IL-10 for the diagnosis of MPP in children were 0.751,0.687,0.784,0.864,0.798,0.672,0.650,and 0.811,and AUC of the combined detection was 0.924.Conclusion Children with TB and MPP have significantly decreased immune function in the early stages of the disease,and abnormal expression of peripheral blood T-lymphocyte subsets and cytokines.Com-pared with TB children,MPP children have lower levels of CD4+,CD4+/CD8+,and higher levels of CD3+,CD8+,IL-10 and IFN-γ,and the T-lymphocyte subsets and cytokine levels are closely related to the changes in the patients'condition.The combined detection of CD3+,CD4+,CD8+,CD4+/CD8+,IL-8,IFN-γ,IL-13 and IL-10 provides a theoretical basis for identifying and diagnosing early MPP in children.
ABSTRACT
Objective:To evaluate the effect of five-element acupuncture on clinical symptoms, brain metabolism and immunoglobulin level in patients with chronic fatigue syndrome.Methods:It was a randomized controlled trial. A total of 94 patients with chronic fatigue syndrome in our hospital from January 2021 to January 2022 were selected and divided into two groups according to the random number table method, with 47 in each group. The conventional western medicine group was treated with conventional western medicine, and the five-element acupuncture group was treated with five-element acupuncture on the basis of the conventional western medicine group. Both groups were treated for 4 weeks. Before and after treatment, the serum levels of interferon-γ (IFN-γ), corticosterone (CORT), IL-2 and 5-hydroxytryptamine (5-HT) were detected by ELISA; the levels of natural killer (NK) cells, CD4 +, CD8 +, IgG and IgM were detected by flow cytometry; the whole body superconducting MRI scanner was used to scan T2 Flair, T2WI and TlWI sequences of the hippocampus, and the spectral curves and the areas under the peak of N-acetylaspartic acid (NAA), creatine (Cr) and choline (Cho) were obtained, and the ratios of Cho/Cr and NAA/Cr were calculated. the fatigue Scale-14 (FS-14) and Fatigue Severity Scale (FSS) were used to evaluate the fatigue state of the patients, and the Depression-Anxiety-Stress Scale-21 (DASS-21) and Beck Anxiety Inventory (BAI) were used to evaluate the anxiety state of the patients. Symptom Checklist 90 (SCL-90) and Somatic and Mental Health Report Score (SPHERE) were used to evaluate the quality of life of patients. And the clinical efficacy was evaluated. Results:After treatment, the levels of IgG, CD4 + and NK in the five-element acupuncture group were significantly higher than those in the conventional western medicine group ( t values were 4.76, 3.65, 6.42, respectively, P<0.01), and the level of IgM, CD8 + was significantly lower than that in the conventional western medicine group ( t values were 7.30, 4.79, P<0.01); the levels of IFN-γ, IL-2 and 5-HT in the observation group were significantly higher than those in the conventional western medicine group ( t values were 7.60, 4.05, 2.79, respectively, respectively, P<0.01), and the level of CORT was significantly lower than that in the conventional western medicine group ( t=6.72, P<0.01); the NAA/Cr levels in the left [(1.10±0.04) vs. (1.05±0.03), t=6.86] and right [(1.18±0.02) vs. (1.21±0.03), t=8.23] hippocampus of the experimental group were significantly higher than those in the conventional western medicine group ( P<0.01), and the Cho/Cr levels in the left [(1.08±0.04) vs. (1.03±0.03), t=5.70] and right [(1.17±0.02) vs. (1.20±0.03), t=5.71] hippocampus of the experimental group were significantly lower than those of the conventional western medicine group ( P<0.01). After treatment, the scores of physical fatigue, mental fatigue and FSS in the five-element acupuncture group were significantly lower than those in the conventional western medicine group ( t values were 8.08, 9.08 and 7.07, respectively, P<0.01). The scores of DASS-21, BAI, SCL-90 and SPHERE in the conventional western medicine group were significantly lower than those in the conventional western medicine group ( t values were 3.63, 5.77, 8.74, 5.92, respectively, P<0.01).The total effective rate was 95.74% (45/47) in the five-element acupuncture group and 82.98% (39/47) in the conventional western medicine group, and there was no significant difference between the two groups ( χ2=2.80, P=0.094). Conclusion:Five-elements acupuncture can improve the expression of T lymphocytes, increase the levels of immunoglobulin and NK, reduce the level of CORT, regulate the brain metabolism of NAA in the left and right hippocampus, improve the clinical symptoms and negative emotions, and improve the clinical efficacy and quality of life in patients with chronic fatigue syndrome.
ABSTRACT
Objective·To investigate the effect of Astragali Radix on T lymphocyte subsets and cytokine expression in Hashimoto's thyroiditis patients with normal thyroid function.Methods·A total of 120 Hashimoto's thyroiditis patients with normal thyroid function and complete data were selected from January 2020 to December 2020 in Jinshan Branch of Shanghai Sixth People's Hospital.The patients were randomly divided into intervention group(n=60)and control group(n=60)by the method of random number table.The treatment plan of the control group was iodine appropriate state diet,and the intervention group was combined with oral Astragali Radix solution(150 mL per time,twice/d)on the basis of the treatment of the control group.The two groups were treated for 6 months.The changes in peripheral blood serum T lymphocyte subsets(CD3+,CD4+,CD8+,and CD4+/CD8+),cytokines[interleukin-2(IL-2),tumor necrosis factor-α(TNF-α),interleukin-6(IL-6),and interleukin-10(IL-10)],hypersensitive C-reactive protein(hs-CRP),erythrocyte sedimentation rate(ESR),thyroid function,autoantibody,liver and kidney function,and other biochemical indexes were compared before and after treatment between the two groups.Adverse reactions were observed during the treatment.The factors influencing the change amplitude of thyroid peroxidase antibody(TPOAb)were analyzed by multifactor linear regression.Results·Finally,118 patients,with 59 cases in each group,were included in the study.After 6 months of treatment,the intervention group showed significant improvements in the proportions of CD4+ T cells,the ratio of CD4+/CD8+,and the levels of IL-2,TNF-α,IL-10,hs-CRP,ESR,TPOAb,and thyroglobulin antibody(TGAb)compared to the values before treatment and in the control group(P<0.05).There were no statistically significant differences on the above indicators before and after treatment in the control group(P>0.05).No serious adverse reactions were observed in the intervention group.Multiple linear regression analysis results showed that the use of Astragali Radix,increase of CD4+ level,increase of CD4+/CD8+ ratio,and decrease of hs-CRP level were influencing factors for the decrease of TPOAb level(β=-0.393,P=0.029;β=-0.513,P=0.010;β=-0.351,P= 0.035;β=0.434,P=0.023).Conclusion·Astragali Radix can improve the levels of CD4+ T cells,CD4+/CD8+ratio,IL-2,TNF-α,IL-6,and IL-10 in Hashimoto's thyroiditis patients with normal thyroid function,and it is safe to use.
ABSTRACT
Objective To explore the application value of lymphocyte subsets combined with various cytokines in the disease progression of elderly patients with corona virus disease 2019(COVID-19).Methods From December 2022 to January 2023,146 elderly patients with COVID-19 diagnosed in the emergency ward of the Eighth Medical Center of PLA General Hospital were selected and divided into two groups according to the prognosis:127 cases in the COVID-19 survival group,19 cases in the COVID-19 death group.In addition,51 osteoporosis patients in geriatric medicine department were collected as control group.The proportion and absolute count of lymphocyte subsets(including T,B and NK cells),and 12 cytokines in plasma(including IL-1β,IL-2,IL-4,IL-5,IL-6,IL-8,IL-10,IL-12p70,IL-17,TNF-α and IFN-γ)were compared between the control group and COVID-19 group,survival group and death group.The receiver operating characteristic(ROC)curve was used to evaluate its prognostic value in elderly patients with COVID-19 infection.Results Compared with the control group:① The proportion of NK cells in COVID-19 group was decreased,while the proportion of B cells was increased,and the differences were statistically significant(Z=-3.386,-4.140,all P<0.01).There was no significant difference in the proportion of T,CD8+T and CD4+T cells,and the differences were not statistically significant(Z=-1.244,-1.770,-0.951,all P>0.05).② The absolute numbers of T,CD8+T,CD4+T,NK and B cells in COVID-19 group were decreased,and the differences were statistically significant(Z=-9.418~-6.539,all P<0.01).③ The concentrations of IL-2,IL-6,IL-1β,IFN-γ,IL-8,IL-17,IL-12P70 and IL-10 in COVID-19 group were all increased,and the differences were statistically significant(Z=-8.851~-1.986,all P<0.05).There was no significant difference in the concentrations of IL-5,IFN-α,TNF-α and IL-4,and the differences were not statistically significant(Z=-0.460~-0.217,all P>0.05).Compared with the survival group:① There was no significant difference in the proportion of T,CD8+T,CD4+T,NK and B cells in the death group(Z=-1.873~-0.422,all P>0.05).② The absolute numbers of T,CD8+T and CD4+T cells in the death group were all decreased,and the differences were statistically significant(Z=-2.667,-2.287,-2.556,all P<0.05),while there was no significant difference in absolute numbers of NK and B cellsm and the differences were not statistically significant(Z=-1.934,-0.532,all P>0.05).③ The concentrations of IL-6,IFN-γ,IL-8,IL-17 and IL-10 in the death group were all increased,and the differences were not statistically significant(Z=-4.211~-2.655,all P<0.05),and there was no significant difference in the concentrations of IL-5,IFN-α,IL-2,IL-1β,IL-12p70,TNF-α and IL-4 the differences were not statistically significant(Z=-1.329~-0.279,all P>0.05).ROC curve analysis for the prognostic value of lymphocyte subsets combined with cytokines in elderly patients with COVID-19 showed that:the areas of total T cells,B cells and NK cells under ROC curve for predicting the prognosis of COVID-19 infection were 0.94,0.80 and 0.93,respectively.The areas of CD4+T cells and CD8+T cells under ROC curve for predicting the prognosis of COVID-19 infection were 0.93 and 0.90,respectively.The areas of IL-6,IFN-γ,IL-8,IL-17 and IL-10 in cytokines under the ROC curve for predicting the prognosis of COVID-19 infection were 0.91,0.71,0.87,0.74 and 0.90,respectively.However,the area of combined lymphocyte subsets and cytokines under ROC curve for predicting the prognosis of COVID-19 infection reached 0.99.Conclusion The immune status of elderly patients with COVID-19 was generally low.Evaluation of immune status has important clinical guidance significance in disease diagnosis,disease observation and prognosis.
ABSTRACT
Although a great deal of research has been done on sepsis in children,its pathogenesis remains controversial and unresolved,one of which is immune dysfunction.Immunosuppression is the core part of immune dysfunction,and it is related to the disease progression,treatment options,and prognosis changes in patients with sepsis.Since T lymphocytes play a crucial role in immune function,they have been regarded as an important indicator for judging changes in immune function in clinical practice.This review explained the mechanism of immunosuppression caused by some T lymphocyte subsets and its clinical application value in the evaluation,prognosis and immune intervention of sepsis.
ABSTRACT
Objective:To explore the relationship between peripheral blood lymphocyte subsets, interleukin-6 (IL-6) levels, and prognosis in patients with viral pneumonia.Methods:A total of 100 patients with viral pneumonia admitted to the Emergency Department of the Chuiyangliu Hospital Affiliated to Tsinghua University from December 2021 to November 2023 were selected and divided into a survival group and a death group. 20 healthy individuals who underwent physical examinations during the same period were selected as the control group. General information of the patients was collected, and peripheral blood lymphocyte subsets and related cytokine IL-6 were detected. Differences in peripheral blood lymphocyte subsets and IL-6 levels among different groups were analyzed, And multiple logistic regression was used to analyze the influencing factors of viral pneumonia disease and death.Results:The proportion of death group with diabetes was significantly higher than that of survival group ( P<0.05). The mortality of patients with diabetes was higher than that of patients without diabetes [60.7%(17/28) vs 12.5%(9/72), P<0.05]. The total number of peripheral blood T lymphocytes, helper/inducible T lymphocytes (Th), CD4 + /CD8 + ratio, B lymphocyte count, and natural killer (NK) cells in patients with viral pneumonia were significantly lower than those in the healthy control group (all P<0.05), and the level of IL-6 was significantly higher than that in the healthy control group ( P<0.05); The total number of peripheral blood T lymphocytes, Th cells, CD4 + /CD8 + ratio, B lymphocyte count, and NK cells in the death group were significantly lower than those in the survival group (all P<0.05), and the level of IL-6 was significantly higher than that in the survival group ( P<0.05). The results of multivariate logistic regression analysis showed that T lymphocyte subsets and IL-6 were all influencing factors for the disease and death of viral pneumonia (all P<0.05). Conclusions:The mortality of viral pneumonia patients with diabetes increased; The absolute number of lymphocyte subsets decreases and the level of IL6 increases in patients with viral pneumonia; The difference in changes between the death group and the survival group is more significant; Early detection of peripheral blood lymphocyte subsets and IL6 levels is beneficial for evaluating the prognosis of patients with viral pneumonia and has certain guiding value for clinical practice.
ABSTRACT
Background and Aims: The host immune system plays an important role in the pathogenesis and defense mechanism of Mycobacterium tuberculosis (Mtb). This study aimed to explore the different changes in the immune system between smear-negative pulmonary tuberculosis (PTB) and smear-positive PTB patients. Materials and Methods: A total of 85 active PTB patients and 50 healthy adults were enrolled. The participants were divided into smear-negative PTB, smear-positive PTB, and control groups. Chest computed tomography (CT) and lymphocyte subgroup counts in peripheral blood were measured in all participants. Results: There were higher numbers of CD4 + T-cells, NK cells, and pulmonary cavities in the smear-positive PTB group, whereas the numbers of B-ells were significantly increased in the smear-negative PTB group. Conclusions: Smear-negative PTB showed fewer pulmonary cavities, mild inflammatory response, lower numbers of immune cells, and higher numbers of B- cells.
ABSTRACT
Objective:To investigate the changes of T lymphocyte subsets in peripheral blood of patients with diffuse large B-cell lymphoma (DLBCL) and its clinical significance.Methods:The clinical data of 99 DLBCL patients admitted to the First Affiliated Hospital of Xiamen University from January 2022 to January 2023 were retrospectively analyzed. T lymphocyte subsets in peripheral blood before and after treatment were detected by using flow cytometry. According to the disease status at the time of blood collection and detection, the patients were divided into the newly-diagnosed DLBCL group (28 cases), and the newly-treated remission DLBCL group (71 cases); and 40 healthy volunteers undergoing the physical examination during the same period were selected as the healthy control group. The proportion and absolute count differences of T lymphocytes and the related subsets in 3 groups were compared. Besides, the correlation among T lymphocyte subsets, the correlation of each subset with international prognostic index (IPI) score and treatment response in newly-diagnosed DLBCL patients were further analyzed.Results:The proportion of CD3 + T cells in newly-diagnosed DLBCL group was decreased compared with that in the healthy control group [(58±14)% vs. (67±7)%, P < 0.05]. The absolute count of CD3 + T cells in both newly-diagnosed group and the newly-treated remission group was reduced compared with that in the healthy control group [(875±483) /μl and (808±553) /μl vs. (1 374±279) /μl, P < 0.001]. The absolute count of CD4 + and CD8 + T cells in newly-diagnosed group was decreased compared with that in the healthy control group [(478±313) /μl vs. (695±154) /μl, (316±181) /μl vs. (525±193) /μl, all P < 0.001]. Both the proportion and absolute count of CD4 + T cells in the newly-treated remission DLBCL group were decreased compared with those in the newly-diagnosed DLBCL group and the healthy control group [(40±14)% vs. (53±14)% and (51±9)%, (313±247) /μl vs. (478±313) /μl and (695±154) /μl, all P < 0.05]. The porportion of CD8 + T cells was increased compared with that in the other two groups [(51±15)% vs. (37±12)% and (38±9)%, all P < 0.001]. Compared with the healthy control group, the effect/memory subsets proportion of regulatory T cell (Treg) and conventional T cell (Tcon) were increased in both newly-diagnosed DLBCL group and the newly-treated remission DLBCL group [(79±16)% and (84±12)% vs. (71±11)%,(72±16)% and (76±14)% vs. (62±13)%, all P < 0.05], and the proportion of CD127 + memory Tcon and CD8 + T cell subsets was reduced [(73±14)% and (66±20)% vs. (85±8)%,(39±15)% and (25±21)% vs. (62±16)%, all P < 0.05]. In newly-diagnosed DLBCL group, the absolute counts of CD3 + T and CD4 + T cells were negatively correlated with the proportion of effector Treg ( r = -0.379, P = 0.049; r = -0.384, P = 0.040, respectively). IPI score of DLBCL patients was correlated with the proportion of CD8 + T cells ( Eta2 = 0.15, P = 0.038). The proportion of CD127 + memory Tcon in patients with non-complete remission was increased compared with that in patients with complete remission after treatment ( P = 0.020). Conclusions:The proportion and absolute count of T lymphocyte cells in peripheral blood of newly-diagnosed DLBCL patients is decreased, and the differentiation state of T lymphocyte cells shows change trend, which is related to the clinical characteristics and treatment response of DLBCL patients. Even if DLBCL patients have achieved treatment remission, T lymphocyte cells are not completely return to the normal.
ABSTRACT
Objective:To investigate the correlation of peripheral blood 25-hydroxyvitamin D3 [25 (OH) D3] level with T cell subsets in multiple myeloma (MM).Methods:The clinical data of 11 newly diagnosed MM patients hospitalized in Heze Municipal Hospital and the First People's Hospital of Jining from June 2019 to June 2021 were retrospectively analyzed, and 8 healthy people were selected as the healthy control group. The patients achieved disease remission after 4 courses of BD (bortezomib + dexamethasone) regimen. High performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) was used to measure the peripheral blood 25-(OH) D3 level in MM patients at initial diagnosis and after 4 courses of treatment, as well as people in the healthy control group. The proportion of peripheral blood helper T cell (Th) 1, Th2, Th17, and regulatory T cells (Treg cells) in CD3 + CD8 + T cells was measured by using flow cytometry. IgA, IgG, IgM, lactic dehydrogenase (LDH), β 2-microglobulin (β 2-MG) levels were analyzed by using fully automatic biochemical analyser. Cytoanalyze was used to detect the hemoglobin level in the peripheral blood. The differences of all indicators in MM patients at initial diagnosis, remission after treatment and the healthy control group were compared. Pearson method was used to analyze the correlation of the peripheral blood 25-(OH) D3 level with T cell subsets and other biochemical indicators in MM patients at initial diagnosis and remission after treatment. Results:Compared with the healthy control group, the peripheral blood 25-(OH) D3 level, Th1-to-Th2 ratio (Th1/Th2), the proportion of Treg cells were all decreased (all P < 0.01), and Th17-to-Treg cells ratio (Th17/Treg) was increased ( P = 0.002). The proportion of Th17 and Th17/Treg in MM patients achieving remission after treatment was higher than that in the healthy control group (all P < 0.05); the proportion of Treg cells in MM patients achieving remission after treatment was lower than that in the healthy control group ( P = 0.010); 25-(OH) D3 level in MM patients achieving remission after treatment was lower than that in the healthy control group, while the difference was not statistically significant ( P = 0.060). The peripheral blood IgM in MM patients at initial diagnosis and those achieving remission after treatment was lower than that in the healthy control group (all P < 0.01); the levels of LDH and β 2-MG in MM patients at initial diagnosis and those achieving remission after treatment was higher than that in the healthy control group (all P < 0.05). The peripheral blood 25-(OH) D3 level in MM patients at initial diagnosis was positively correlated with the proportion of Th1, Th1/Th2 ( r values 0.89, 0.60, all P < 0.05), and negatively correlated with the proportion of Th17 and Th17/Treg ( r values -0.61, -0.75, all P < 0.05). After treatment, there was no correlation of the proportion of Th1, Th2, Th17, Treg, Th1/Th2, Th17/Treg with peripheral blood 25- (OH) D3 level for patients achieving remission ( r values were -0.36, -0.45, -0.10, 0.10, 0.19, 0.03, all P > 0.05). IgM, LDH, β 2-MG was negatively correlated with 25- (OH) D3 level in the peripheral blood of MM patients at initial diagnosis ( r values were -0.76, -0.71, -0.62, all P < 0.05); while there was no correlation of 25-(OH) D3 level with IgA, IgG, IgM, LDH, β 2-MG, hemoglobin for patients achieving remission after treatment ( r values were -0.36, 0.19, -0.09, 0.47, 0.47, -0.11, all P > 0.05). Conclusions:MM patients show the decreased peripheral blood 25-(OH) D3 level, the increased Th17 and the decreased Treg cells; 25-(OH) D3 level is related to the imbalance of Th1/Th2/Th17/Treg, which suggests that 25-(OH) D3 may be related to the development, progression, prognosis and abnormal immune responses in the body of MM.
ABSTRACT
Objective To study the short-term efficacy of combined cervical metastatic lymph nodes hy-perthermia during locally advanced nasopharyngeal carcinoma induction chemotherapy and concurrent chemo-radiotherapy and its effect on peripheral blood lymphocyte subsets.Methods Sixty patients with pathological-ly diagnosed nasopharyngeal carcinoma in this hospital from July 2021 to July 2022 were collected as the study subjects and divided into the observation group(induction chemotherapy+concurrent chemoradiotherapy combined with hyperthermia)and control group(induction chemotherapy+concurrent chemoradiotherapy),30 cases in each group.The general information,short term efficacy,EB virus(EBV)DNA level,adverse reac-tions occurrence,peripheral blood lymph cell subsets and hot shock protein 90α(HSP90α)were recorded and compared between the two groups.Results Compared with the control group,the objective remission rate in the observation group was higher(100.0%vs.90.0%),the EBV positive rate after induction chemotherapy was lower(20.0%vs.46.7%),the occurrence rate of ≥3 grade radiation dermatitis was higher(30.0%vs.6.7%),the level of natural killer(NK)cells after radiotherapy was increased[(25.89±5.53)%vs.(19.18±6.41)%],the HSP90α level after treatment was increased[(91.19±9.18)ng/mL vs.(67.22± 11.02)ng/mL],and the differences were statistically significant(P<0.05).Compared with before treatment,the levels of CD3+,CD4+,CD4+/CD8+after radiotherapy in the observation group were decreased,the levels of CD3+,CD4+,CD4+/CD8+in the control group were decreased,but the differences between the two groups were not statistically significant(P<0.05),while the proportion of peripheral blood NK cells in the experi-mental group was significantly increased compared with the control group(P<0.05).Conclusion Hyper-thermia canimprove the local control of nasopharyngeal carcinoma cervical lymph node metastasis and improve the immune function.
ABSTRACT
Objective:To analyze the correlation between peripheral blood lymphocyte subsets and unex-plained repeated implantation failure(URIF),and to investigate its predictive value for the diagnosis of URIF.Methods:A total of 156 patients with URIF who underwent treatment in the Department of Reproductive Medi-cine,The Second Affiliated Hospital of Kunming Medical University from October 1,2019 to December 1,2021 was selected as the URIF group.Meanwhile,age-matched 41 women with a history of healthy delivery in the past one year were selected as the control group.The percentages of peripheral blood lymphocyte subsets in the two groups were measured by flow cytometry,and the results were statistically analyzed.Receiver operating charac-teristic(ROC)curve was used to evaluate the predictive effect of lymphocyte subsets on URIF.Results:Com-pared with the control group,the percentage of CD3 + CD4 +[(34.03±7.26)%vs.(36.79±6.35)%,P = 0.017]、CD3 +HLA-DR +[(2.60±2.28)%vs.(3.60±2.39)%,P =0.017]、CD3 + CD16 + CD56 +[(1.24±1.04)%vs.(2.62±2.57)%,P<0.000]and CD4 +/CD8 +(1.37±0.48 vs.1.57±0.51,P =0.023)were sig-nificantly increased in URIF group,and were related to the increase of previous failure times to a certain extent.ROC analysis showed that CD3 + CD4 +>35.35%(AUC 0.624),CD3 + HLA-DR +>2.35%(AUC 0.669),CD3 +CD16 +CD56 +>1.86%(AUC 0.660)and CD4 +/CD8 +>1.26(AUC 0.628)could be used as an inde-pendent predictor for the diagnosis of URIF.Among the pair-wise combined indexes,CD3 + HLA-DR + combined with CD3 + CD16 + CD56 + had the highest prediction accuracy(AUC 0.739,Sensitivity 73.3%,Specificity 68.3%).The combination of the four indicators had the highest accuracy(AUC0.767,Sensitivity 68.6%,Specifici-ty 73.2%).Conclusions:There is immune dysfunction in patients with URIF,and the imbalance of peripheral blood lymphocyte subsets may be an important factor leading to embryo implantation failure.CD3 +CD4 +、CD4 +/CD8 +、CD3 +HLA-DR +and CD3 +CD16 +CD56 +could be used as independent indicators for the diagnosis of abnormal peripheral blood lymphocyte subsets in patients with URIF.The combination of them improves the accuracy of prediction,and it has a positive reference significance for dynamic monitoring and early intervention of URIF pa-tients in the process of assisted reproduction technology.
ABSTRACT
Objective:To assess the changes of peripheral blood NK cells in patients with systemic lupus erythematosus (SLE) following belimumab and classic therapy.Methods:From January 2020 to March 2022, peripheral lymphocyte subsets were detected by flow cytometry in SLE patients treated with Belimumab and classic therapy. The duration of treatment was 24 weeks. A total of 40 treated SLE patients were enrolled. The lymphocyte subsets in healthy donors were used as normal control group. Paired sample t-test, rank-sum test, Spearman correlation and generalized linear mixed model were used for statistical analysis. Results:In contrast to healthy subjects, the numbers of NK cells in SLE patients before treatment were significantly decreased [276.0 (179.8, 384.0) cells/μl vs. 61.4 (43.0, 105.1) cells/μl; Z=-7.32, P<0.001], although that after treatment was higher than that before treatment [61.4 (43.0, 105.1) cells/μl vs. 107.7 (72.5, 186.5) cells/μl; Z= -3.22, P<0.001]. Generalized linear mixed model results showed that the increase in serum levels of C3 ( t= -2.94, P=0.006) and NK cells ( t=-2.25, P=0.031) were associated with a decrease of anti-dsDNA antibody titers. The cutoff value of elevated counts of NK cells after treatment was equal or more than 38.5 cells/μl with a sensitivity of 61.9% and a specificity of 81.2%. Compared with those with elevated counts of NK cells ≤38.5 cells/μl, patients with elevated counts of NK cells >38.5 cells/μl had a bigger difference anti-dsDNA antibody [49.2 (0.2, 207.2) vs. 156.2 (19.8, 260.7); Z=-1.55, P=0.120] and a bigger difference of SLEDAI-2000[4.5 (0.0, 10.0) vs. 13.0 (4.5, 18.0); Z=-2.52, P=0.012]. Conclusion:The change in the numbers of NK cells may serve as biomarkers for evaluating the therapeutic responses of SLE. Combinatory approaches employing belimumab and classic therapy may control SLE disease by increasing the number of NK cells
ABSTRACT
Objective:By detecting the species and distribution of fecal flora in patients with pSS, we investigated the relationship between the alterations of the gut microbiome and its metabolic characteristics with peripheral lymphocyte subsets, and their potential role in the occurrence and development of pSS.Methods:A total of 101 pSS patients who were hospitalized in the department of rheumatology of the Second Hospital of Shanxi Medical University, and 101 age and sex-matched healthy control (HC) in the health check-up center of Shanxi Provincial People′s Hospital from January 2019 to January 2020 were enrolled for 16s rDNA-Amplicon sequencing. The statistical analysis was performed in R software 4.0.3. The Alpha diversity were compared by Wilcoxon test, and Beta diversity were compared by ANOSIM analysis between pSS patients and HC. The difference flora was analyzed by t test. The levels of peripheral lymphocyte subsets pSS patients were detected by flow cytometry. Then the relationship between characteristic flora and clinical indicators such as lymphocyte subsets, erythrocyte sedimentation rate, C-reactive protein, unstimulated whole saliva and stimulated whole saliva were analyzed using Pearson′s correlation analysis. Results:Patients with pSS exhibited a significant reduction in the richness (Chao1, ACE) and diversity (Shannon,Simpson) of gut microbiota compared with those of HC, and there was statistical significant difference in gut microbiota composition (ANOSIM, r=0.09, P=0.001). At the phylum level, the relative abundance of Firmicutes decreased and the relative abundance of Proteobacteria increased in pSS patients. At the genus level, the proportion of Escherichia-shigella ( P<0.001), Lactobacillus ( P<0.001), Bifidobacterium ( P<0.001), Subdoligranulum ( P<0.001), Alistipes ( P<0.001) and [ Eubacterium]_ coprostanoligenes (P=0.002) were increased. The proportion of Faecalibacterium ( P<0.001), Prevotella ( P<0.001), Roseburia ( P<0.001), Megamonas ( P<0.001), Agathobacter ( P<0.001), Lachnospira ( P<0.001), Lachnospira_NK4A136 ( P=0.006), [ Eubacterium] _eligens ( P<0.001) were significantly reduced. PICRUST analysis showed significant enrichment of amino acid metabolism taurine and hypotaurine metabolism ( P<0.001), fatty acid metabolism such as propanoate metabolism ( P<0.001), glutathione metabolism ( P=0.002), lipoic acid metabolism ( P=0.003), lipopolysaccharide biosynthesis and biosynthesis of siderophore group nonribosomal peptides ( P=0.005) and Aminobenzoate degradation ( P=0.002) in patients with pSS. The Pearson correlation results showed that there were significant different in the abundances of the key gut microbiota between the HC and pSS groups, and they were closely related to unstimulated whole saliva, the absolute number of Treg cells and Th17 cells. Conclusion:The dysbiosis and metabolism changes of the pSS intestinal microbiota may contribute to immune homeostasis imbalance, and may be involved in the occurrence and development of pSS.
ABSTRACT
Objective:To investigate the clinical value of dynamic detection of lymphocyte subsets and blood cell counts in management of patients with lupus nephritis (LN).Methods:The clinical data of 65 patients with primary LN admitted in Affiliated Hospital of Qingdao University from January 2015 to April 2021 were retrospectively analyzed. According to the stage of disease progression and medications used,LN patients were classified into primary active phase,post-induction therapy phase,and maintenance therapy phase. The changes in lymphocyte subsets were monitored,and the relationship of lymphocyte subsets and blood cell count ratios with lupus activity and infection events was evaluated.Results:The decrease of CD4 +T lymphocyte and NK cell counts were negatively correlated with the activity of systemic lupus erythematosus (SLE)( r=-0.67,-0.33, P<0.01),while CD8 +T lymphocyte,B cell counts,neutrophil/lymphocyte ratio (NLR),platelet/lymphocyte ratio (PLR),and monocyte/lymphocyte ratio (MLR) were positively correlated with the SLE activity( r=0.38,0.26,0.34,0.26,0.29, P<0.05). The area under ROC curve (AUC) of CD4 +T lymphocyte count in predicting the occurrence of infection in LN patients was the highest (0.89); taking 247.50 cell/μl as cutoff value,the sensitivity and specificity were 81.25% and 87.50%,respectively. The combination of CD4 +T lymphocyte with CRP increased the predicting value for the occurrence of infection. Conclusion:Dynamic detection of blood lymphocyte subsets and blood cell counts can reflect SLE activity and the occurrence of infection in LN patients. Among these indicators the CD4 +T lymphocyte has the highest predictive value for the occurrence of infection,and the combination of the CD4 +T lymphocyte count with CRP level can further improve the predicting value.
ABSTRACT
ObjectiveTo explore the effect of Tongxie Yaofang on the immune microenvironment of colorectal cancer in mice under chronic stress and the underlying mechanism. MethodA total of 40 male SPF BABL/C mice were randomized into normal group, stress group, Tongxie Yaofang group (13.65 g·kg-1), and Tongxie Yaofang-stress group (13.65 g·kg-1), with 10 in each group. Chronic restraint stress was induced in mice and administration (ig) of Tongxie Yaofang began after 7 days of stress. On the 14th day, forced swim and tail suspension tests were used to examine the behavioral changes of mice after stress and the subcutaneous colorectal tumor was implanted in each group of mice. The effect of this prescription on the body mass and tumor volume of mice was observed. After the last administration, mouse serum and tumor samples were collected. The content of T lymphocytes (CD3+, CD4+, CD8+, and CD4+/CD8+) in tumor was detected by immunohistochemistry and flow cytometry and levels of corticosterone (CORT) in peripheral blood, and interleukin (IL)-2, interferon-γ (IFN-γ), IL-6, and IL-10 in the serum were determined by enzyme-linked immunosorbent assay (ELISA). The protein expression of inhibitor of nuclear factor-κB(IκB) kinase α/β (IKKα/β), nuclear factor-κB (NF-κB)α (IκBα), NF-κB p65, and phosphorylated (p)-NF-κB p65 was measured by Western blot. ResultCompared with the normal group, the stress group had large tumor volume (P<0.05), low content of CD3+, CD4+, and CD4+/CD8+ (P<0.05, P<0.01), high content of CD8+, low content of T helper 1 (Th1)-secreted IFN-γ (P<0.05), high content of T helper 2 (Th2)-secreted IL-10 (P<0.05) and CORT (P<0.05), high protein expression of p-NF-κB p65, NF-κB p65, and IKKα/β (P<0.05), and low protein expression of IκBα (P<0.05). Compared with the normal group, the Tongxie Yaofang group showed slow tumor growth, high content of CD3+, CD4+, and CD4+/CD8+ (P<0.01), low content of CD8+ (P<0.05), high content of Th1-secreted IL-2 and IFN-γ (P<0.05), low content of Th2-secreted IL-6 and IL-10 (P<0.05), low content of CORT, low protein expression of p-NF-κB p65, NF-κB p65, and IKKα/β (P<0.05), and high protein expression of IκBα (P<0.01). Tongxie Yaofang-stress group demonstrated slower tumor growth, higher content of CD3+, CD4+, and CD4+/CD8+ (P<0.01), smaller content of CD8+ (P<0.05), higher content of IL-2 and IFN-γ (P<0.05), lower content of IL-6, IL-10 (P<0.05), and CORT (P<0.05), lower protein expression of p-NF-κB p65, NF-κB p65, and IKKα/β (P<0.05,P<0.01), and higher protein expression of IκBα (P<0.01) than the stress group. ConclusionTongxie Yaofang can delay the growth of colorectal cancer under chronic stress and alleviate the deterioration of the immune microenvironment, possibly by inhibiting NF-κB signaling pathway, regulating the function of T lymphocyte subsets, and thus suppressing the secretion of pro-inflammatory factors.
ABSTRACT
OBJECTIVE@#To observe the clinical efficacy of bamboo-based medicinal moxibustion for chronic fatigue syndrome (CFS), and to preliminarily explore its action mechanism.@*METHODS@#Sixty-four patients with CFS were randomly divided into a moxibustion group (32 cases, 1 case dropped off, 1 case excluded) and an acupuncture group (32 cases, 2 cases dropped off). The patients in the moxibustion group were treated with bamboo-based medicinal moxibustion, while the patients in the acupuncture group were treated with routine acupuncture. Both groups were treated once a day, 6 days as a course of treatment with 1 day interval, for a total of 2 courses of treatment. Before treatment, 1 and 2 courses into treatment and in the follow-up of 14 days after treatment, the fatigue scale-14 (FS-14) and somatic and psychological health report (SPHERE) scores were observed in the two groups. Before and after treatment, the contents of CD+3, CD+4, CD+8 of peripheral blood T lymphocyte subsets were measured and CD+4/CD+8 ratio was calculated; the clinical efficacy of the two groups was compared.@*RESULTS@#Compared before treatment, the FS-14 and SPHERE scores in the two groups were decreased 1 and 2 courses into treatment and in the follow-up (P<0.01), and the FS-14 and SPHERE scores in the moxibustion group were lower than those in the acupuncture group (P<0.01, P<0.05). Compared before treatment, the contents of CD+3, CD+4 and CD+4/CD+8 ratio in the moxibustion group were increased after treatment (P<0.01). There was no significant difference of CD+3, CD+4, CD+8 and CD+4/CD+8 ratio between before and after treatment in the acupuncture group (P>0.05). After treatment, the contents of CD+3 and CD+4 in the moxibustion group were higher than those in the acupuncture group (P<0.05). The total effective rate was 93.3% (28/30) in the moxibustion group, which was higher than 73.3% (22/30) in the acupuncture group (P<0.05).@*CONCLUSION@#Bamboo-based medicinal moxibustion could improve the physical and mental fatigue symptoms and psychological status in patients with CFS. Its effect may be related to regulating the contents of CD+3, CD+4 of peripheral blood T lymphocyte subsets and CD+4/CD+8 ratio.
Subject(s)
Humans , Moxibustion , Fatigue Syndrome, Chronic/therapy , Acupuncture Therapy , Physical ExaminationABSTRACT
Objective:To investigate the difference of lymphocyte subsets between elderly patients with rheumatoid arthritis and non-elderly patients and its clinical significance.Methods:A total of 124 patients with rheumatoid arthritis in Affiliated Hospital of Nantong University from January, 2017 to December, 2019 were enrolled.The patients were divided into elderly group(≥60 years old, 34 cases)and non-elderly group(<60 years old, 90 cases). Rheumatoid arthritis activity(DAS-28)scoring was performed for each patient.Peripheral blood mononuclear cells(PBMCs)were extracted by Ficoll density centrifugation.Lymphocytes were labeled and detected by 18-color flowcytometry with more than 30 fluorescent antibodies.Results:DAS-28 scoring showed that the disease activity score of the elderly group(4.56±1.89)was higher than that of the non-elderly group(3.37±1.49)( t=3.633, P<0.001). Flow cytometry showed that MAIL%T(mucus-associated lymphoid tissue T cell subset)( Z=-2.798, P=0.005), Tn%CD8 T cells(initial CD8 T cells)( Z=-2.179, P=0.029), VD2% T(Vδ2+ T, γδT cell subtype)( Z=-2.806, P=0.005), PD1-CD28-%Th( Z=-2.050, P=0.040)and IGM+ D-%B( Z=-2.376, P=0.017)were lower in the elderly group than in the non-elderly group.While, CD45+ CD27+ %CD8 T cells( Z=-3.069, P=0.002), abT%T cell(αβT cells)( Z=-2.103, P=0.035), CD27-CD28+ %T cells( Z=-2.341, P=0.019), ASC%PBMC( Z=-2.341, P=0.019)and ASC%CD19+ ( Z=-2.000, P=0.046)subgroup expression were higher in the elderly group than in the non-elderly group. Conclusions:The disease activity of elderly patients with rheumatoid arthritis is significantly higher than that of younger patients.The expressions of abT%T and CD4% abT in effector T cells of elderly patients with rheumatoid arthritis are higher than those of younger patients, while the expression of VD2% T is lower.The expression level of CD45RA+ CD27+ %CD8 T with cytotoxic effect is higher; However, the expression level of Tn%CD8 T in naive cells is lower.
ABSTRACT
Objective:To probe into the potential prognostic value of lymphocyte subsets in gastric cancer.Methods:This study included patients who underwent radical gastrectomy for gastric cancer from Aug 2014 to Dec 2016. The immunological differences was analyzed in different infiltration patterns. The overall survival of patients was analyzed by Kaplan-Meier method and Log-rank test. COX regression was performed to assess independent prognostic factors of the patients, and finally constructed nomogram.Results:The median number of peripheral CD4 and CD19 cells in infiltration pattern c was 750 (94-2 504) cells/μl and 186 (17-820) cells/μl; the median number of peripheral CD4 and CD19 cells in infiltration pattern a was 802 (203-2 071) cells/μl and 213 (5-948) cells/μl, the number of peripheral CD4,CD19 cells in infiltration pattern c was lower than that in infiltration pattern a, with statistically significant differences (CD4: Z=-3.061, P=0.002; CD19: Z=-2.016 , P=0.044). CD19 lymphocytes ( P=0.023) were associated with infiltration pattern a, CD8 lymphocytes ( P=0.027) were associated with infiltration pattern b, and CD4 lymphocytes ( P=0.026) were independent risk factors associated with the prognosis of infiltration pattern c. A nomogram can be constructed to evaluate the prognosis of patients. Conclusion:There are differences in the number of peripheral lymphocyte subsets in patients with different INF types. A nomogram can be constructed from lymphocyte subsets and clinicopathological features to assess patient prognosis.
ABSTRACT
Objective:To provide data reference for using Chinese rhesus macaques as research model by studying the immunophenotype and function of peripheral blood lymphocytes in Chinese rhesus macaques.Methods:By optimizing antibody clones and fluorescent colors, the lymphocyte subset assay and T cell function assay panels were determined. Then the panels were used to analyze the proportion of T, B, NK and other cell subsets in peripheral blood mononuclear cells (PBMCs) in 15 healthy Chinese rhesus monkeys, and the ability of T cells to secrete cytokines after non-specific stimulation.Results:Two multi-color flow cytometry analytic panels were established. Panel 1 could simultaneously detect a variety of lymphocyte subsets, including cytotoxic T lymphocytes, follicular helper T cells, regulatory T cells, B cells and NK cells. Panel 2 could detect the functions of multiple T cell subsets and the expression of immune checkpoint moleculars. The mean percentages of T, B, NK, Tfh, Treg, CD16 + NK and CD56 + NK cells in PBMCs of the Chinese rhesus macaques were (75.32±7.73)%, (13.22±7.50)%, (0.88±0.48)%, (0.73±0.27)%, (0.75±0.43)%, (47.87±22.35)% and (10.69±12.41)%. After non-specific stimulation, the proportion of CD4 + T cells secreting IL-2 and TNF-α was higher than that of CD8 + T cells, and the proportion of CD8 + T cells secreting CD107a and IFN-γ was higher than that of CD4 + T cells, while the proportion of CD4 + and CD8 + T cells secreting IL-17A was low. Conclusions:This study established a multi-color flow detection scheme that could simultaneously detect multiple cellular surface molecules and cytokines at the single cell level and could accurately and comprehensively analyze the immune cell subsets, functions and the immune checkpoint molecules in peripheral blood of Chinese rhesus macaques, providing a new experimental method and basic data for the development of vaccines and drugs against infectious diseases.
ABSTRACT
ObjectiveTo study the effect of Bazi Bushen capsules on delaying the aging process of naturally aging mice and its mechanism. MethodThe mice were randomly divided into four groups according to their body weight, namely, aging group, low-dose Bazi Bushen capsules group (1 g·kg-1), high-dose Bazi Bushen capsules group (2 g·kg-1), and rapamycin group (0.002 g·kg-1). The debilitating signs were detected by behavioral tests and the weakness index was measured. The percentages of spleen T and B lymphocytes, effector T cells (TE), memory T cells (TM), naive T cells (TN), helper T cells (Th), cytotoxic T cells (Tc) ,Th1 cells, Th2 cells, and regulatory T cells (Treg) were determined by flow cytometry. Cell proliferation and the cell counting kit-8 (CCK-8) assay was used to detect the proliferation of lymphocytes in mice. The hematoxylin-eosin (HE) staining was used to observe the histopathological changes in the mouse spleen. The expression of cyclin-dependent kinase inhibitor 2A (p16) and cyclin-dependent kinase inhibitor 1A (p21) was detected by immunohistochemistry (IHC). The mRNA expression of senescence-related proteins p16 and p21 was determined by real-time fluorescence quantitative polymerase chain reaction (Real-time PCR), and the inflammatory cytokines, including tumor necrosis factor-α (TNF-α), interferon-γ (IFN-γ), interleukin (IL)-1β, IL-2, IL-4, IL-10, and IL-12 p70, in peripheral blood of mice were detected by Luminex. ResultAs compared with the aging group, mice in the Bazi Bushen capsules and rapamycin groups showed significantly improved debilitating signs and reduced weakness index scores (P<0.05, P<0.01), increased proportions of T cells, TN cells, Tc cells, Th2 cells, and Treg cells in the spleen, decreased proportions of TE cells, TM cells, Th cells, Th1 cells (P<0.05, P<0.01), and increased proliferation of splenic lymphocytes (P<0.05, P<0.01). In the Bazi Bushen capsules and rapamycin groups, clear structure of the red and white marrow marginal zone was observed in the spleen of mice, the area of the white marrow was increased, and the area of the red marrow was correspondingly decreased. The protein and mRNA expression of aging-related proteins p16 and p21 in the spleen was decreased (P<0.01), the levels of serum pro-inflammatory cytokines IL-1β, IL-2, IL-12 p70, IFN-γ, and TNF-α levels were decreased, and the levels of IL-4 and IL-10 were increased (P<0.05, P<0.01) in the Bazi Bushen capsules and rapamycin groups as compared with the aging group. ConclusionBazi Bushen capsules have the effect of regulating the debilitating signs of natural aging mice, regulating the immune homeostasis and inflammation level of the body, and reducing cell aging.