Objective To investigate the effect of high mobility group box-1 protein (HMGB1) on inter-leukin-2 (1L-2) and interleukin-2 receptor α (IL-2α) expressions in human T lymphocytes and its potential regulat-ing mechanism in vitro. Method Human T lymphocytes were isolated and suspended, the cells were cultured with 20 μg/mL phytohemagglutinin (PHA) in 5% CO_2 at 37 ℃, recombinant human HMGB1 (rhHMGB1, 0, 10, 100, 1000 ng/mL) was added with the PHA and cultures were centrifuged at 12 and 48 hours for cell collect-ing. Reverse transcription polymerase chain reaction (RT-PCR) amplification was perfomed to determine gene ex-pressions of IL-2, IL-2Rα. IL-2, sIL-2R protein levels in cell culture supematants were measured by ELIZA. Re-sults After coincubated with rhHMGB1 (10, 100, and 1000 ng/mL) for 12 hours, IL-2 levels in cell culture su-pernatants respectively were 0 . 064 ± 0. 017 μg/L, 0.076±0.033 μg/L, and 0.061 ±0.02 μg/L, which were significantly higher compared with the untreated cells (0.045±0.011 μg/L, P < 0.05 or P < 0.01). Mean-while, IL-2 mRNA expression was markedly up-regulated following rhHMGB1 stimulation in various doses (F = 4.6872, P < 0.01). At 48 bourn, however, both IL-2 mRNA expression and protein production tended to de-crease along with an increased dose of dd-IMGB1 stimulationn. IL-2/sIL-2R ratio in 1000 ng/mL rhHMGB1 was markedly lower than that in 10 ng/ml rhHMGB1 (0.036±0.015 vs.0.055±0.017, P <0.05), together with down-regulation of IL-2Rα mRNA expression(P <0.01). Conclusions These data indieated that HMGBI could marked influence the IL-2/IL-2R expression in human T lymphocytes. With the increase in stimulating doses and prolongation of time, HMGBI might down-regulate T cell-mediated immune response of human lymphocytes.

Child ; Chondrosarcoma, Mesenchymal ; diagnosis ; drug therapy ; pathology ; Diagnosis, Differential ; Humans ; Male ; Pelvic Neoplasms ; diagnosis ; drug therapy ; pathology ; Tomography, X-Ray Computed ; Treatment Outcome

Objective To investigate the immunological activity change of regulatory T cells (Treg) and discuss its significance in the outcomes of patients with multiple organ dysfunction syndrome (MODS) and severe burn. Methods A total of 106 patients with total burn surface area (TBSA) larger than 30% were included in the study and randomly divided into three groups according to the burn area: Group Ⅰ (TBSA of 30%-49%, n = 41), Group Ⅱ (TBSA of 50% -69%, n = 34) and Group Ⅲ (TBSA of 70%-99%, n = 31). According to the development of MODS, patients were divided into MODS group (n =21) and non-MODS group (n =85). The patients with MODS were further divided into non-survival group (n = 16) and survival group (n = 5) based on their outcomes. Healthy volunteers were served as normal control (n = 25). Peripheral blood samples were collected at days 1,3,7, 14 and 21 postburn. The immunomagnetic separation technique was applied to separate and purify CD4+ CD25+Tregs in peripheral blood, and phenotypes (CTLA-4) were analyzed by flow cytometry and the contents of interleukin-10 released in the supernatants were determined by ELISA. Results Expression of CTLA-4 and level of IL-10 were significantly increased in burn patients compared with normal control group, with statistical differences. The expression of CTLA-4 and level of IL-10 were significantly increased in patients with severe burns at all time points. The expression of CTLA-4 and level of IL-10 in MODS group were much higher than those in non-MODS group at days 3-21 postburn (P ＜ 0.01). Among the MODS patients, the expression of CTLA-4 and level of IL-10 in the survival group were obviously lower than those in the non-survival group at days 3-21 postburn (P ＜ 0.05 or P ＜ 0.01). Conclusions After severe burn injury, expressions of the markers on CD4 + CD25 + Treg surface and secretion of cytokines produced by CD4 + CD25 + Tregs show significant difference in patients with different born areas, MODS development and survival state. CD4 + CD25 + Treg may play an important role in the pathogenesis of immunoregulation, MODS and mortality of burn patients through secretion of inhibitory cytokines.

Objective To study the induction of integrin molecules mediated by CYR61 in human peripheral blood mononuclear cells(PBMC). Methods A recombinant expression plasmid containing full length of human CYR61 labeled with human IgG Fc fragment was constructed and identified by DNA sequencing.COS7 was used as host cell for identification of secretary CYR61 expression,confirmed by Western blot method.The commercial lipofectin was adopted for recombinant plasmid transfection into PBMC.Real-time PCR was ultilized to analyze expression patterns of CYR61 and integrin molecules in transfected PBMC. Results The insert sequence was correct in the recombinant plasmid.Western blot test showed that CYR61 protein secreted into culture supernatant or was in COS7 cytoplasm.The recombinant plasmid was transfected into PBMC stimulated with phytohemagglutinine(PHA) to induce CYR61 expression and secretion.The results demonstrated that exogenous CYR61 transcribed rapidly after being incubated with PHA and reached the peak after 24 h.But the expression dropped down quickly to a very low level after 48 h.Simultaneously,integrin molecules expressed just after CYR61 transcription.In the set of integrin molecules tested in the study,?v,?M,?3 and ?5 expression were higher than the other integrin molecules(P

Objective To analyze the relationships between the drinking water fluoride and bone mineral density (BMD), and serum osteocalcin (BGP) and to explore the BMD and serum BGP as significant early screening biomarkers for fluorosis especially for early bone damage in endemic fluorosis areas. Methods Wamiao (severe endemic fluorosis area, as fluoride exposed group) and Xinhuai (non endemic fluorosis area, as control group) Village were selected in 2006. One hundred and fouty-six objects were chosen from 2 villages (103 in Wamiao, 43 in Xinhuai). The sex, age, body height, body weight, drinking water fluoride in each object's household well, BMD, and serum BGP were investigated, and the dose-response relationships were analyzed between the drinking water fluoride and BMD, and serum BGP. CurveExpert 1.3 Software was used to fit the dose-response relationships between the rate of abnormal BMD, the rate of abnormal serum BGP, and the drinking water fluoride. Results The levels of drinking water fluoride in males' and females' families in fluoride exposed group were [(2.38±0.68), (2.62±0.91 )mg/L] significant higher than that in control group [(0.35±0.08), (0.36±0.07)mg/L], the difference being statistically significant(t values were 14.27 and 11.08,and P<0.01, respectively). BMD in males in fluoride exposed group [(0.78±0.07)g/cm2] was significant lower than that in control group[(0.83±0.08)g/cm2], the difference being statistically significant (t=2.37,P<0.05). Serum BGP in males and females in fluoride exposed group [(4.17±0.67), (4.11±0.57) μg/L] were significant higher than that in control group [(1.48±0.40), (1.44±0.39)μg/L], the difference being statistically significant (t values were 17.64 and 19.40, and P<0.01, respectively]. BMD in the group with drinking water fluoride≥2.92 mg/L[(0.66±0.15 )g/cm2] was significant lower than that in the group with drinking water fluoride<0.42 mg/L [(0.76±0.12)g/cm2], the difference being statistically significant (P<0.01). The levels of serum BGP in the groups with the drinking water 0.42-,2.05-, ≥.92 mg/L[(3.83±1.07), (4.22±0.72), (3.99±0.63) μg/L] were significant higher than that in the group with the drinking water<0.42 mg/L [(1.44±0.37) μg/L], the difference being statistically significant (P<0.01). The equation for the dose-response relationship between the drinking water fluoride and the rate of abnormal BMD was y=(0.284-0.058x)-1.260, r=0.999 94; and y=100.05/(1+78.62e-4.5x), r=0.999 99 for the drinking water fluoride and the rate of abnormal serum BGP. Conclusions There were significant dose-response relationships between drinking water fluoride and BMD and serum BGP. It indicated that BMD and BGP might be considered as early screening biomarkers for endemic fluorosis, especially for the bone damage.

Objective This study was performed to investigate the effect of high mobility group box-1 protein (HMGB 1) on immune function of human T lymphocytes in vitro and explore its potential role in cell-mediated immune dysfunction.Methods Fresh blood was obtained from healthy adult volunteers and peripheral blood mononuclear cells (PBMCs) were isolated,then rhHMGB 1 was added to PBMCs.Four-color flow cytometric (FCM) analysis was used for the measurement of intracellular cytokine including interleukin Results (1) Different stimulating time and dosage of rhHMGB 1 did not alter the number of IFN-a positive cells (Th 1).rhHMGB 1 stimulation provoked a dose-dependent and time-dependent increase in Th2 subset and decrease in ratio of Th 1 to Th2.(2) Compared with the untreated cells,when the cells were coincubated with rhHMGB 1 (10-100ng/ml) for 12 hrs,protein release of IL-2 and sIL-2R were significantly up-regulated.At 48 hrs,in contrast,protein production was relatively lower in cells after exposure to 100-1000 ng/ml rhHMGBI.Conclusions These findings demonstrated that HMGB1 has a dual influence on immune functions of human T lymphocytes.

OBJECTIVETo investigate the immunological effect of PM2.5 on cytokine production in female Wistar rats.

METHODSFemale Wistar rats were given 0.3 mg, 0.75 mg, 2 mg, 5 mg of PM2.5 per 0.5 mL saline, respectively. Saline was used as the negative control. TNF-alpha and IL-6 levels in the branchoalveolar lavage were measured by ELISA, and mRNA expression levels in lung tissue were detected by RT-PCR. Alveolar macrophages were collected for testing phogacytic function.

RESULTSExposure to PM2.5 stimulated TNF-alpha production in a dose-dependent manner (P < 0.05), However, no statistically significant difference was found. No time-dependent change in TNF-a and IL-6 production was found. TNF-alpha and IL-6 mRNA expressions were induced by PM2.5-exposure. The phagocytic rate (PR) was significantly decreased by PM2.5 treatment.

CONCLUSIONPM2.5 exposure increases inflammation response of the lung in a dose-dependent manner. Moreover, tissue injury induced by PM2.5 may be related to altered production of cytokines. PMz2.5 may impair the phagocytic activity of alveolar macrophages.

Animals ; Bronchoalveolar Lavage Fluid ; Cytokines ; biosynthesis ; genetics ; metabolism ; Enzyme-Linked Immunosorbent Assay ; Female ; Macrophages, Alveolar ; metabolism ; Particle Size ; Phagocytosis ; RNA, Messenger ; genetics ; Rats ; Rats, Wistar ; Reverse Transcriptase Polymerase Chain Reaction

Objective:To evaluate the assistant role of manifestations under tracheoscopy in the diagnosis of invasive pulmonary aspergillosis (IPA) in severe patients.Methods:A retrospective study was conducted. The patients with suspected IPA admitted to intensive care unit (ICU) of Affiliated Hospital of Binzhou Medical College from January 2015 to December 2019 were enrolled. The diagnosis, clinical diagnosis and suspected diagnosis were made according to the grading criteria of Guidelines for the diagnosis and treatment of invasive fungal infection in severe patients (2007). Those who met the criteria were enrolled in the IPA group, and those who did not meet the criteria or other pathogens were enrolled in the non-IPA group. The general data of the patients were collected, and the changes of tracheal and bronchial mucosa under tracheal microscope before and after treatment were recorded, as well as the results of galactomannan (GM) test and aetiology culture of bronchoalveolar lavage fluid (BALF). The baseline, bronchoscopy and pulmonary CT manifestations and their dynamic changes were compared in each group. Results:A total of 142 patients with suspected IPA were finally enrolled. Among them, 12 were pathologically proven IPA, 77 were probable IPA, 22 were possible IPA, and 31 were undefined IPA. Of the 142 patients, 60 had typical manifestations of mucosal injury under bronchoscopy, including 7 proven IPA patients (58.3%), 52 probable IPA patients (67.5%), and 1 possible IPA patient (4.5%), but none undefined IPA patient. The patients undergoing lung CT scan were 12 proven IPA patients (100%), 73 probable IPA patients (94.8%), and 21 possible IPA patients (95.5%), respectively. Most of the Chest CT showed patchy or strip density increasing and other non-specific manifestations. There were 3 proven IPA patients (25.0%), 7 probable IPA patients (9.0%), and 0 possible IPA patient (0%) who had typical IPA CT manifestations (halo sign and cavity or crescent sign). Among the patients of proven IPA and probable IPA (89 cases), there were a total of 35 cases with endoscopic airway mucosal injury and tracheoscopy reexamination ≥ 3 times. All the 35 patients received anti-aspergillus treatment, among which 16 survived and 19 died. Among the 16 patients who survived, the microscopic appearance of mucosal injury was gradually reduced and the clinical manifestations were gradually improved. Of the 19 patients who died, 16 had deteriorated endoscopic airway mucosal injury.Conclusions:The specific manifestations of severe patients with bronchial mucosal injury are of great significance in the diagnosis of IPA. In the case of severe patients who cannot receive pathological examination or chest CT in time, dynamic observation of the changes of airway mucosal injury is a simple auxiliary method to discover the changes of patients' condition in time, evaluate the effect of antifungal therapy and the prognosis of IPA.

OBJECTIVETo observe the short-term efficacy and radiotherapy induced acute reactions of sarcandra glabra (SG) combined chemoradiotherapy for treating patients with local advanced nasopharyngeal carcinoma.

METHODSTotally 100 nasopharyngeal carcinoma phase III - IVa patients were randomly assigned to the control group and the observation group, 50 patients in each group. All patients received chemoradiotherapy. Patients in the observation group took SG decoction at the daily dose of 20 g.

RESULTSTotally 98 patients completed the treatment. There was no statistical difference in the complete remission rate, the partial remission rate, 1-year and 2-year total survival rates, or disease free progression rate between the two groups. The occurrence rate of II - III degree radioactive oral mucositis and dry mouth were obviously lower in the observation group than in the control group, showing statistical difference (P <0.05).

CONCLUSIONSG combined chemoradiotherapy could attenuate radiotherapy induced acute adverse reactions in treating advanced nasopharyngeal carcinoma.

Adult ; Aged ; Carcinoma ; Chemoradiotherapy ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Male ; Middle Aged ; Nasopharyngeal Neoplasms ; pathology ; therapy ; Neoplasm Staging ; Phytotherapy

Adult ; Female ; Humans ; Sarcoma, Myeloid ; diagnosis ; Vulvar Neoplasms ; diagnosis