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.

Objective:To analyze the results of lymphocyte subsets and 12 plasma cytokines in patients with tuberculosis by flow cytometry and to evaluate their diagnostic efficacy in these patients.Methods:This is a retrospective case-control study. A total of 128 patients with evidence of tuberculosis disease or clinically confirmed tuberculosis who were admitted to the 8th Medical Center of PLA General Hospital from January 2022 to December 2023 were included. According to the location of mycobacterium tuberculosis infection, the patients were divided into the pulmonary tuberculosis group (83 cases) and the extrapulmonary tuberculosis group (45 cases), and 100 healthy age-and sex matched people who underwent health check up during the study period were selected as the control group. Flow cytometry was used to detect peripheral blood lymphocyte subsets and 12 plasma cytokines [including 10 pro-inflammatory factors: interleukin (IL)-5, interferon (IFN)-α, IL-2, IL-6, IL-1β, IFN-γ, IL-8, IL-17, IL-12P70, Tumor necrosis factor (TNF)-α, and two anti-inflammatory factors: IL-4, IL-10] in participants of all groups. Spearman correlation method was used to analyze the correlation between lymphocyte subsets and cytokines, binary Logistic regression was used to screen the TB related factors, and receiver operating curve (ROC) was used to evaluate the diagnostic efficacy of TB related factors.Results:Compared with the control group, the absolute number of CD3 +T lymphocytes, CD3 +CD8 +T lymphocytes, CD3 +CD4 +T lymphocytes, NK cells and B cells were lower in pulmonary tuberculosis group and extrapulmonary tuberculosis group (all P<0.05). Except for IL-1β, the levels of other 11 cytokines are all significantly higher in the pulmonary tuberculosis group (all P<0.01), and the levels of IL-6, IFN-γ, IL-17, TNF-α, IL-4 and IL-10 were significantly higher in extrapulmonary tuberculosis group (all P<0.05). Compared with extrapulmonary tuberculosis group, the level of IL-8 was higher in pulmonary tuberculosis group ( P=0.026). Spearman correlation analysis showed that IL-6, IFN-γ and IL-8 were negatively correlated with the absolute numbers of CD3 +T lymphocytes, CD3 +CD8 +T lymphocytes, CD3 +CD4 +T lymphocytes, NK cells and B cells (IL-6: R2=-0.30, -0.28, -0.32, -0.26, -0.28; IFN-γ: R2=-0.36, -0.31, -0.37, -0.25, -0.36; IL-8: R2=-0.14, -0.13, -0.16, -0.14, -0.22; all P<0.05), IL-10 was negatively correlated with the absolute number of CD3 +CD4 +T lymphocytes, NK cells and B cells ( R 2=-0.14, -0.19, -0.21, all P<0.05); Binary Logistic regression analysis showed that IL-6, IFN-γ, IL-8 and IL-10 were the related factors of tuberculosis ( OR=1.809, 1.136, 0.910, 2.218, all P<0.05), ROC curve analysis showed that the AUC of IL-6, IFN-γ, IL-8 and IL-10 in the joint diagnosis of tuberculosis was 0.845, the sensitivity was 0.766, and the specificity was 0.820. Conclusion:The lower absolute number of lymphocyte subsets and cytokine levels in patients with pulmonary tuberculosis and extrapulmonary tuberculosis indicate that their immune function is in a low state, and the higher levels of pro-inflammatory factors (IL-6, IFN-γ, IL-8) and anti-inflammatory factor (IL-10) indicates the higher inflammatory status, and evaluation of these 4 cytokines has satisfactory diagnostic efficacy for tuberculosis.

Objective:This study aims to establish an early warning diagnosis model for acute exacerbation of chronic obstructive pulmonary disease (AECOPD) and to provide a simple, rapid, and accurate auxiliary diagnosis basis for clinical practice.Methods:The sample bank of subjects (patients admitted to the Eighth Medical Center of the PLA General Hospital from September 10, 2021, to July 25, 2023) was constructed, including the model establishment cohort [SCOPD group 49, 42 males and 7 females, (69.71±11.16) years old; AECOPD group 53, 49 males and 4 females, (72.60±10.19) years old] and the model validation cohort [SCOPD group 35, 28 males and 7 females, (69.97±10.40) years old; AECOPD group 35, 33 males and 2 females, (71.43±9.67) years old]. Fasting peripheral blood samples were collected, and the expression levels of IL-5, IL-17A, and IFN-α were detected by flow cytometry. Different expression levels were analyzed by Mann-Whitney U test. Binary logistic regression analysis was used to screen the related risk factors of COPD patients in acute exacerbation. The diagnostic efficacy of the model was evaluated by the receiver operating characteristic (ROC) curve.Results:The levels of IL-5 [1.64 (0.60, 2.86) pg/ml], IL-17A [1.42 (0.88, 2.29) pg/ml], and IFN-α [0.91 (0.59, 1.81) pg/ml] in the SCOPD group were significantly decreased compared with the AECOPD group IL-5 [4.68 (2.34, 9.40) pg/ml, Z=-5.033, P<0.001], IL-17A [2.33 (1.59, 4.62) pg/ml, Z=-3.919, P<0.001], IFN-α [2.83 (0.91, 3.75) pg/ml, Z=-4.127, P<0.01] in the cohort of model establishment. The results of binary logistic regression analysis between SCOPD and AECOPD groups showed that IL-5, IL-17A, and IFN-α were independent risk factors for acute exacerbation of patients with COPD ( P<0.05). And the regression equation is Y=-2.861+0.364×IL-5+0.385×IL-17A+0.445×IFN-α. The AUC value of IL-5, IL-17A, IFN-α and combined detection was 0.866 ( P<0.001). Compared to the SCOPD group and the AECOPD group in the cohort of model validation, the receiver operating characteristic (ROC) curve showed that the combined model of three (AUC=0.858, P<0.001) could be used to diagnose the AECOPD. And the Kappa value was 0.773( P<0.05). Conclusion:The combined detection of IL-5, IL-17A, and IFN-α has high diagnostic efficacy for patients with acute exacerbation of COPD. This method provides a new potential tool for the clinical diagnosis of AECOPD and has the value of further exploration and optimization, promotion, and application.

【Objective】 To investigate the current status of incision sites to obtain intact specimens in laparoscopic nephrectomy by urologists in China, so as to provide reference for the standardized procedure. 【Methods】 During Jun.20, 2021 and Jul.4, 2021, more than 20 000 urologists in a WeChat group were surveyed with a questionnaire. The general data, incision sites and related complications were statistically analyzed. 【Results】 A total of 601 valid questionnaires were collected, covering urologists from 31 provinces, autonomous regions and municipalities. Surgical approaches: 68 urologists chose trans-abdominal approach, 432 chose posterior abdominal space approach, 101 chose both surgical approaches. Incision sites: 97 urologists chose lumbar transverse incision, 202 chose dorsal oblique incision of the waist, 119 chose ventral oblique incision, 93 chose the paramedian incision, 112 chose the lower abdominal oblique incision (Gibson), 11 chose the transverse lower abdominal incision (Pfannenstiel), 7 chose the median incision of the lower abdomen, 2 chose the median incision in the upper abdomen, 15 chose axillary midline direct incision; 399 chose to cut off the muscles, and 202 chose not to. Complications: 232 urologists reported pain after 2 weeks, 369 reported no pain; 325 reported numbness after 2 weeks, 276 reported no numbness; 66 reported incisional hernia, 535 reported no hernia. 【Conclusions】 Chinese urologists tend to choose retroperitoneoscopic nephrectomy and waist incision to obtain intact specimens. Transperitoneal laparoscopic nephrectomy has a variety of incisions for intact specimens. There is no standardized incision sites to obtain intact specimens.

Objective To evaluate the effect of bone marrow mesenchymal stem cell (BMSC) on the expression of interleukin (IL)-10 and tumor necrosis factor (TNF)-α in mice with ischemia-reperfusion acute kidney injury (IR-AKI). Methods All mice were randomly divided into the sham operation group (control group), ischemia-reperfusion injury group (IRI group) and BMSC treatment group (BMSC group), with 6 mice in each group, respectively. The renal function and pathological changes of mice were detected. The cell apoptosis of renal tissues of mice was determined. The expression levels of serum IL-10 and TNF-α of mice were quantitatively measured. The mouse BMSC was randomly divided into the control and hypoxia-reoxygenation groups (IRI group), and the expression levels of IL-10 and TNF-α in cell supernatant were determined. Results The renal structure of mice was normal in the control group, severe damage was observed in the IRI group, and mild damage occurred in the BMSC group. Compared with the control group, the renal tissue injury scores were significantly higher in the IRI and BMSC groups (both P < 0.05). Compared with the IRI group, the renal tissue injury score was significantly lower in the BMSC group (P < 0.05). Compared with the control group, the levels of serum creatinine (Scr) and blood urea nitrogen (BUN) were remarkably up-regulated in the IRI group, and the level of BUN was significantly up-regulated in the BMSC group (all P < 0.05). Compared with the IRI group, the levels of Scr and BUN were significantly down-regulated in the BMSC group (both P < 0.05). In the IRI group, the quantity of apoptotic cells in the renal tissues was considerably higher than those in the BMSC and control groups, and the quantity of apoptotic cells in the BMSC group was significantly higher than that in the control group (all P < 0.05). Compared with the control group, the levels of serum IL-10 and TNF-α were significantly up-regulated in the IRI group, whereas the level of serum TNF-α was significantly down-regulated and the level of serum IL-10 was significantly up-regulated in the BMSC group (all P < 0.05). Compared with the IRI group, the levels of serum IL-10 and TNF-α were significantly down-regulated in the BMSC group (both P < 0.05). The levels of IL-10 and TNF-α in the cell supernatant did not significantly differ between the IRI and control groups (P=0.080、0.627). Conclusions BMSC infusion may reduce the incidence of renal IRI and inflammation, probably via the mechanism of down-regulating TNF-α expression rather than up-regulating IL-10 expression.

Objective To investigate the dynamic changes of peripheral blood lymphocyte subsets and their correlation with renal function in recipients with stable graft status after renal transplantation. Methods Forty-five recipients who underwent renal transplantation for the first time and had stable graft function within postoperative 6 months were selected. The proportion and absolute value of lymphocyte subsets were detected by flow cytometry (FCM) in 180 peripheral blood samples from recipients at 15 d, 1, 3 and 6 months after renal transplantation. The dynamic changes of lymphocyte subsets with the extension of postoperative time and their correlation with serum creatinine (Scr) and blood urea nitrogen (BUN) were analyzed. Results The Scr levels did not significantly differ at 4 time points after renal transplantation (all P > 0.05). The BUN levels significantly differed between 15 d and 1 month after renal transplantation, and between 1 and 3 months after renal transplantation (P=0.002, P=0.001). The proportion of CD3⁺CD8⁺T cells, CD3⁺CD4⁺T cells, natural killer (NK) cells and CD4/CD8 ratio at postoperative 15 d significantly differed from those at 1 month after operation (P=0.009, P=0.004, P < 0.001, P=0.004). The proportion of B cells significantly differed between 15 d and 1 month, and between 1 and 3 months after renal transplantation (both P < 0.001). The absolute values of CD3⁺T cells, CD3⁺CD8⁺T cells, CD3⁺CD4⁺T cells and NK cells at postoperative 15 d significantly differed from those at 1 month after renal transplantation (P=0.001, P=0.002, P=0.003, P < 0.001). The absolute values of CD3⁺CD8⁺T cells significantly differed between 3 and 6 months after operation (P=0.015). The absolute value of B cells at 1 month after renal transplantation significantly differed from that at 3 months after renal transplantation (P=0.001). The proportion and absolute value of lymphocyte subsets were not significantly correlated with the Scr level (both P > 0.05). The proportion and absolute value of CD3⁺CD8⁺T cells and NK cells were negatively correlated with BUN (P < 0.001-0.05), whereas the proportion of CD3⁺CD4⁺T cells and B cells was positively correlated with the BUN level (P < 0.001-0.05). The absolute value of CD3⁺T cells was negatively associated with the BUN level (P < 0.05). Conclusions T cells and NK cells in the lymphocyte subsets of stable recipients raise to the stable state within 1 month after renal transplantation, whereas B cells decrease to stable state within 3 months renal transplantation. The dynamic changes of lymphocyte subsets are correlated with the BUN level.

Objective To explore the effect of umbilical cord mesenchymal stem cells with positive human leukocyte antigen(HLA)-G on inducing the production of regulatory T cells(Treg) in vitro.Methods Umbilical cord mesenchymal stem cells were isolated from umbilical cord of neonates. PEGFP-N1-HLA-G plasmid was transfected into the human umbilical cord mesenchymal stem cells by liposome transfection, as PEGFP-N1-HLA-G group. PEGFP-N1 empty vector plasmid was transfected into the human umbilical cord mesenchymal stem cells, as PEGFP-N1 group. The human umbilical cord mesenchymal stem cells without empty vector under the same conditions were set as blank control group. Markers of the umbilical cord mesenchymal stem cells were detected using flow cytometry. The expression of HLA-G protein in each group of cells was identified by Western Blot. After mixed-culturing with CD4+T cells in peripheral blood of healthy subjects for 24 h and 48 h, the proportion of CD4+CD25+Foxp3+Treg in total T cells of each group was detected by flow cytometry. Results CD45, CD34 and HLA-DR presented negative expression on umbilical cord mesenchymal stem cells, while CD29, CD44 and CD105 presented positive expression. HLA-G protein could be expressed in the PEGFP-N1-HLA-G group, which had statistically significant difference compared with the blank control group and PEGFP-N1 group (both P<0.01). After PEGFP-N1-HLA-G group and CD4+T cells were mixed-cultured for 24 h and 48 h, CD4+CD25+Foxp3+Treg accounted for (15.3±1.9)% and (14.3±2.1)% of the total T cells respectively, both of which presented statistically significant difference compared with the blank control group and PEGFP-N1 group (all P<0.05). Conclusions Umbilical cord mesenchymal stem cells with HLA-G gene modified can effectively induce the production of CD4+CD25+Foxp3+Treg in vitro.

Objective To investigate the change rules and its significance of erythrocytes surface molecule CD35 , CD58 and CD59 expression in recipients infected with cytomegalovirus (CMV)after renal transplantation. Methods Eighty-two recipients undergoing allogeneic renal transplantation were selected and divided into the negative (n=21 )and positive CMV groups (n=61 )based on the qualitative detection of CMV-pp65 antigen in peripheral blood. According to the results of CMV-pp65 (+)leucocyte count,all 61 patients in positive CMV group were further divided into low (n=55)and high active infection subgroups (n =6 ). Healthy adults were recruited into the normal control group (n =30 ). The expression levels of CMV-pp65 antigen,erythrocytes surface molecule CD35,CD58 and CD59 were measured by flow cytometry. Results Compared with normal control group,the expression levels of erythrocytes surface molecule CD35 , CD58 and CD59 in the positive CMV group were significantly down-regulated,and the CD35 and CD59 expression in the negative CMV group were considerably down-regulated (all P<0. 05 ). Compared with negative CMV group,the expression levels of CD58 and CD59 in the positive CMV group were significantly down-regulated (both P<0. 05 ). The expression levels of CD35 and CD59 in the high active infection subgroup were significantly lower than those in the low active infection subgroup (both P<0. 05 ). Conclusions The more severe active CMV infection after renal transplantation,the lower expression of erythrocytes surface molecule CD35,CD58 and CD59,hinting that red cell immune dysfunction is probably involved with active CMV infection.

BACKGROUND:Studies have reported that taurine has a certain therapeutic effect on the disease of various systems, such as nervous system, cardiovascular system, immune system and digestive system. The liver is the main place, also the important target organ, of taurine metabolism. Therefore, the relationship between taurine and hepatopathy has become a hot topic in recent years. OBJECTIVE:To investigate the influence of taurine on superoxide dismutase and malondialdehyde expression in the liver tissue of rat models of liver fibrosis induced by carbon tetrachloride. METHODS:Thirty male C57B/L rats of SPF grade were randomly and evenly divided into blank control, model and taurine groups. Rats in the blank control group were intraperitonealy injected with 100% peanut oil of 1 mL/kg, twice a week, in total 10 weeks. Rats in the model group were intraperitonealy injected with peanut oil of 1 mL/kg containing 20% carbon tetrachloride, twice a week, in total 10 weeks. Rats in the taurine group were intraperitonealy injected with peanut oil of 1mL/kg containing 20% carbon tetrachloride, twice a week, in total 10 weeks, and were intragastricaly administered taurine of 500 mg/kg per day starting from the 3rd week til the 10th week. RESULTS AND CONCLUSION:Compared with the blank control group, the serum levels of hyaluronic acid, laminin, typeⅢ procolagen, typeⅣ colagen, alanine aminotransferase, aspartate aminotransferase were significantly increased (P < 0.05), the level of superoxide dismutase in the liver tissue was lowered (P < 0.05), the level of malondialdehyde in liver tissue was significantly increased (P < 0.05), and liver index was increased (P < 0.05) in the model group. Pathological examination showed that there were necrosis of liver cels, fat vacuoles, fibrous tissue hyperplasia and inflammatory cel infiltration in the rats of the model group. Compared with the model group, the serum levels of hyaluronic acid, laminin, typeⅢ procolagen, typeⅣ colagen, alanine aminotransferase, aspartate aminotransferase were significantly lowered (P < 0.05), the level of superoxide dismutase in the liver tissue was significantly increased (P < 0.05), the level of malondialdehyde in the liver tissue was significantly lowered (P < 0.05), and liver index was significantly decreased (P < 0.05) in the taurine group. Pathological examination showed that there were no inflammatory cel infiltration, fat vacuoles, and fibrous tissue deposition in the liver tissue. The results indicate that taurine can decrease the contents of superoxide dismutase and malondialdehyde, and relieve the degree of liver fibrosis induced by carbon tetrachloridevia exerting its antioxidative effects.

Objective To observe the changes of serum homocysteine (Hcy)level of renal transplant recipients before and after renal transplantation,and assess the correlation between serum Hcy level and graft function.Methods Thirty-three recipients were included into the transplantation group,who underwent renal allograft transplantation in the Organ Transplant Institute of the 309 th Hospital of People's Liberation Army and had renal function recovered stably from January 2013 to June 2014.And 65 patients who were confirmed as chronic renal failure (CRF)by clinical examinations were included into the CRF group and 30 healthy people were included into the control group.A retrospective cross-sectional study was conducted on all of these subjects.Serum Hcy,serum creatinine (Scr)and blood urea nitrogen (BUN)levels of these three groups were compared.Serum Hcy and Scr levels of the transplantation group were continuously monitored before transplantation and at 3,7,14 and 21d after transplantation.The correlation between the changes of serum Hcy levels and the renal function before and after transplantation was assessed.Results Serum Hcy level of the CRF group was (25 ±10)μmol/L,which was significantly higher than (9 ±4)μmol/L of the control group and (15 ±9)μmol/L of the transplantation group in stable period,with statistical significance (all in P <0.001).Serum Hcy level of the transplantation group was significantly higher than that of the control group(P <0.001).Scr level of the CRF group,the transplantation group and the control group was(708 ±302)μmol/L, (98 ±23)μmol/L and (72 ±18)μmol/L,respectively.Scr level of the CRF group was significantly higher than those of the transplantation group and the control group (all in P <0.001).BUN level of the CRF group, the transplantation group and the control group was (18.1 ±5.9)mmol/L,(10.9 ±5.3)mmol/L and (4.9 ± 1.3) mmol/L, respectively. BUN level of the CRF group was significantly higher than that of the transplantation group and the control group (all in P <0.001),and BUN level of the transplantation group was significantly higher than that of the control group (P <0.001).With the improvement in renal function after transplantation,Scr and serum Hcy levels of the transplantation group deceased gradually.At 14 d after transplantation,Hcy level decreased to the minimum of (15 ±5)μmol/L.Compared with (25 ±10)μmol/L before transplantation,the difference had statistical significance (P <0.05).Within 14 d after transplantation, serum Hcy level of the transplantation group was positively correlated with Scr level (r =0.761,P <0.05). Conclusions Serum Hcy level of the renal transplant recipients is correlated with the graft function.The combined detection of serum Hcy and renal function index has certain guiding significance in the prevention of hyperhomocysteinemia and the early assessment of graft function.