1.Study on effect and mechanism of salidroside on cognitive ability of Abeta1-40 -induced Alzheimer's disease model rats.
Jia ZHANG ; Ligang SONG ; Weina KONG ; Tiemei SHAO ; Xin WEN ; Pei LIU ; Xiqing CHAI
China Journal of Chinese Materia Medica 2012;37(14):2122-2126
OBJECTIVETo study the effect and possible impact mechanism of salidroside on cognitive ability of Alzheimer's disease (AD) model rats induced by amyloid beta peptide (Abeta1-40).
METHODAbeta1-40 was injected into bilateral hippocampus to create the AD model. Afterwards, different doses of salidroside (25, 50, 75 mg x kg(-1)) were orally administered for 21 days. Rats' learning and memory abilities were detected by Morris water maze testing system. The levels of the superoxide dismutase (SOD), malondialdehyde (MDA), and the expression of nuclear factor-kappaB (NF-kappaB), inducible nitric oxide synthase (iNOS) and receptor for advanced glycation end products (RAGE) protein in hippocampus were also detected by different methods.
RESULTThe place navigation test showed longer escape latency, low frequency of platform quadrant crossing per unit time, damage in learning capacity, significant decrease in SOD acivity in hippocampus, notable increase in MDA content, NF-kappaB, iNOS and RAGE protein expressions in rats. Salidroside (50, 75 mg x kg(-1)) significantly alleviated the impairments of learning and memory ability. The activity of SOD increased in salidroside (50 droside group compared with that of the Alzheimer's disease group (P < 0.01).
CONCLUSIONSalidroside may treat Alzheimer's disease by inhibiting the oxidative stress.
Alzheimer Disease ; drug therapy ; Amyloid beta-Peptides ; toxicity ; Animals ; Cognition ; drug effects ; Disease Models, Animal ; Glucosides ; pharmacology ; therapeutic use ; Male ; Maze Learning ; drug effects ; NF-kappa B ; metabolism ; Nitric Oxide ; physiology ; Phenols ; pharmacology ; therapeutic use ; Rats ; Receptor for Advanced Glycation End Products ; Receptors, Immunologic ; analysis ; Superoxide Dismutase ; metabolism
2.Production pressures among anaesthesiologists in Singapore.
Jia Xin CHAI ; Shin Yuet CHONG
Singapore medical journal 2018;59(5):271-278
INTRODUCTIONProduction pressure is the pressure on personnel to prioritise production ahead of safety. We assessed the prevalence of production pressures among anaesthesiologists in Singapore.
METHODSA random online survey was conducted among local anaesthesiologists. Questions were asked about attitudes to production pressures in the work environment, occurrence of situations involving unsafe actions, and rating of the intensity of external and internal sources of pressure.
RESULTSDemographically, our respondents were largely similar to all anaesthesiologists in Singapore and were fairly distributed across various tertiary hospitals. Nearly half (44.5%) had witnessed production pressures, with a colleague pressured to conduct anaesthesia in an unsafe manner. Such events included pressure from surgeons to proceed for elective surgery in patients without adequate optimisation, pressure to employ anaesthetic techniques that surgeons wanted, having to source for operating rooms to finish the surgeon's list, and being misled regarding surgical time. Over half (52.3%) made errors in clinical judgement due to excess workload. A heavy elective list workload was significantly associated with proceeding with patients despite lack of appropriate support, making changes to practices to avoid delaying the start of surgery and sourcing for operating rooms to finish the surgeon's list (p < 0.05), and being pressured to proceed with patients that the anaesthesiologist would otherwise have cancelled (p < 0.01). The need to avoid delaying the start of surgery and reduce turnover time between patients were the top-ranked internal and external pressures, respectively.
CONCLUSIONProduction pressure is prevalent among anaesthesiologists in Singapore and is correlated with a heavy workload.
Anesthesia ; methods ; Anesthesiologists ; Anesthesiology ; methods ; Elective Surgical Procedures ; Humans ; Operating Rooms ; Patient Safety ; Prevalence ; Singapore ; Stress, Physiological ; Surveys and Questionnaires ; Tertiary Care Centers ; Treatment Outcome ; Workload
3.Construction and significance of recombinant hF9 minigene and its stable nonsense mutant cell lines.
Gang WANG ; Bo-Wen JIANG ; Lin-Hua YANG ; Xin NIE ; Chen-Liang JIA ; Jing LIU ; Quan SHEN ; Bao-Feng CHAI
Journal of Experimental Hematology 2013;21(2):422-425
This study was purposed to construct the recombinant hF9 minigene and its stable nonsense mutant cell lines, and to investigate its significance. Minigene hF9 was cloned into the mammalian expression vector pCMV-Tag3B; a nonsense mutant containing a premature termination codon (PTC) in the 121(st) amino acid residue was obtained by PCR site-directed mutagenesis; minigene hF9 and nonsense mutant were respectively transfected into HepG2 cells with G418 treatment to get stable HepG2-WT and HepG2-N cell lines. The results confirmed that the minigene hF9 and nonsense mutant were constructed successfully. The gene of interest was amplified by RT-PCR from the stable cell lines, and the minigene hF9 was expressed in the stable cell lines. It is concluded that the recombinant hF9 minigene and its stable nonsense mutant cell lines are constructed successfully. The cell lines can be used to screen the drugs treating the nonsense mutation-caused hemophilia according to PTC read-through approaches.
Cell Line, Tumor
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Codon, Nonsense
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Factor IX
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genetics
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Genetic Vectors
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Hemophilia B
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genetics
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Humans
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Recombinant Proteins
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genetics
4.Evaluation of surgical outcomes using imaging study for patients with slow transit constipation complicated with outlet obstruction.
Xiang-dong LIU ; Wei ZHANG ; Bao-hai YU ; Jia-ning ZHAO ; Bin CHAI ; Xin-yun FAN ; Shi-peng ZHAO ; Fa ZHAO
Chinese Journal of Gastrointestinal Surgery 2010;13(10):737-740
OBJECTIVETo explore the clinical significance of multiple radiography of the pelvis in the evaluation of surgical outcomes for patients with slow transit constipation complicated with outlet obstruction.
METHODSPatients with slow transit constipation complicated with outlet obstruction were diagnosed by multiple radiography of the pelvis after screening using colon transit study. Surgery was performed according to the cause of the obstruction. Anorectal angle and the locations of perineum, pelvic peritoneum, and bladder were assessed by multiple radiography of the pelvis one month after surgery. The changes in locations of pelvic organs were assessed and the imaging appearance after the release of obstruction was observed.
RESULTSA total of 48 patients were included. Rectocele repair, partial mucosectomy with rectopexy, and hysteropexy were performed. All the patients were followed up with a mean length of 19(6-58) months. Excluding 2 patients who had no symptomatic improvement, the mean bowel movements was 1.9 times per day in the remaining 46 patients(95.8%). Preoperative anorectal angle at the squeezing phase was(128.09±13.82) degree and the difference between squeezing and resting phase was (11.14±12.58) degree, while the postoperative angle was (180.26±9.98) degree and the difference(20.01±13.11) degree(P<0.05). Preoperative location of the perineum at the squeezing phase was(-2.05±0.83) cm and the difference was(2.23±0.78) cm, while postoperative location was (-0.50±1.13) cm and the difference was (2.18±1.04) cm(P<0.05). Preoperative location of the pelvic peritoneum at the squeezing phase was(4.91±1.32) cm and the difference was (1.32±0.89) cm, while postoperative location was (2.62±2.53) cm and the difference was (3.28±0.68) cm (P<0.05). Preoperative bladder location at the squeezing phase in patients with urological symptoms was (3.92±2.51) cm and the difference was(1.39±1.27) cm, while postoperative location was (2.15±1.55) cm and the difference was (1.98±1.54) cm(P<0.05).
CONCLUSIONMultiple imaging of the pelvis provides objective evidence in the evaluation of surgical outcomes for patients with chronic slow transit constipation complicated with outlet obstruction.
Adult ; Aged ; Constipation ; complications ; diagnosis ; surgery ; Diagnostic Techniques, Digestive System ; Female ; Humans ; Intestinal Obstruction ; complications ; diagnosis ; surgery ; Male ; Middle Aged ; Treatment Outcome ; Young Adult
5.Change in T cell-mediated immunity and its relationship with high mobility group box-1 protein levels in extensively burned patients.
Ning DONG ; Bo-Quan JIN ; Yong-Ming YAO ; Yan YU ; Yu-Jue CAO ; Li-Xin HE ; Jia-Ke CHAI ; Zhi-Yong SHENG
Chinese Journal of Surgery 2008;46(10):759-762
OBJECTIVETo investigate the change in T cell-mediated immunity and its relationship with plasma high mobility group box-1 protein (HMGB1) levels in severely burned patients.
METHODSThirty-five extensively burned patients (> 30% total body surface area) were included in this study, and were divided into MODS group (n = 13) and non-MODS group (n = 22). The blood samples were collected on post burn days 1, 3, 5, 7, 14, 21 and 28. The plasma levels of HMGB1 were measured by using ELISA, and T lymphocyte proliferation response and its IL-2 production ability in peripheral blood were determined too. In addition, the ratio of CD4+/CD8+ T cells were detected by using flow cytometry.
RESULTSPlasma HMGB1 levels were markedly elevated on post burn day 1 in severely burned patients, and HMGB1 level was significantly higher in MODS group than in non-MODS group (P < 0.05). Lymph proliferation response and IL-2 production of T cells in peripheral blood, and the ratio of CD4+/CD8+ T cells in MODS group were markedly lower than those in non-MODS group on post burn days 1, 14, 21 and 28 (all P < 0.05). It indicated that plasma HMGB1 levels were negatively correlated to T cellular immune function parameters, including lymphocyte proliferation response, IL-2 production, and the ratio of CD4+/ CD8+ T cells in extensively burned patients (all P < 0.05).
CONCLUSIONSExtensive burns could lead to T cellular immune dysfunction, which appears to be associated with the development of MODS. HMGB1, as an important late mediators of inflammation, might be involved in the pathogenesis of suppression of T cell-mediated immunity in these patients.
Adolescent ; Adult ; Burns ; blood ; complications ; immunology ; Female ; HMGB1 Protein ; blood ; Humans ; Immunity, Cellular ; immunology ; Male ; Middle Aged ; Multiple Organ Failure ; etiology ; immunology ; T-Lymphocytes ; immunology
6.The clinical significance of changes in immunological function of T lymphocyte in severe burn patients with sepsis.
Ning DONG ; Yong-Ming YAO ; Yu-Jue CAO ; Li-Xin HE ; Yan YU ; Jia-Ke CHAI ; Zhi-Yong SHENG
Chinese Journal of Burns 2007;23(2):84-87
OBJECTIVETo observe the immunological function changes in T lymphocyte in severe burn patients with sepsis, and to explore its relationship with sepsis.
METHODSFifty-nine burn patients with burn surface exceeding 30% TBSA were enrolled in the study, and they were divided into sepsis group (S, n =43) and non-sepsis group (NS, n = 16). The peripheral venous blood samples of the patients in both groups were collected on 1, 3, 5, 7, 14, 21 and 28 post-burn days (PBD). The T lymphocyte proliferation ability and the interleukin-2 (IL-2) level in both groups were observed and the correlation between them were analyzed. The percentage of CD3+/CD4+ T lymphocytes and its apoptosis rate were determined by flow cytometry and the correlation between them was analyzed.
RESULTSCompared with that in NS group, the proliferation ability of T lymphocyte and the level of IL-2 were significantly decreased in patients in S group on 1, 14, 21, and 28 PBD (P < 0.05 or P < 0.01). The inhibition of T lymphocyte proliferation was positively correlated to the low level of IL-2 production in burn patients (r = 0.82, P < 0.01). The percentage of CD3+/CD4+ T lymphocytes in S group were obviously lower than that in NS group on 1, 5, 14, 21, 28 PBD, whereas on opposite tendency in the apoptosis rate of CD3+ CD4+ T lymphocytes were found at the same time (P < 0.05 or P < 0.01). The percentage of CD3+/CD4+ T lymphocytes was negatively correlated to apoptosis rate of T lymphocytes (r = -0.66, P < 0.05).
CONCLUSIONThe immunological function of T lymphocyte in severely burn patients with sepsis is depressed persistently. Apoptosis of T lymphocyte may participate in the pathological process of cell immunological disorder induced by sepsis.
Adolescent ; Adult ; Burns ; blood ; immunology ; pathology ; Cell Proliferation ; Female ; Humans ; Interleukin-2 ; blood ; Lymphocyte Count ; Male ; Middle Aged ; Sepsis ; blood ; immunology ; T-Lymphocytes ; cytology ; immunology ; Young Adult
7.Clinical significance of changes in quantitative expression of human leukocyte antigen DR in severely burned patients.
Ning DONG ; Yong-ming YAO ; Yu-jue CAO ; Li-xin HE ; Jia-ke CHAI ; Shan XU ; Zhi-yong SHENG
Chinese Journal of Surgery 2007;45(11):766-769
OBJECTIVETo investigate the clinical significance of kinetic changes in quantitative expression of human leukocyte antigen DR (HLA-DR) in severely burned patients.
METHODSThe blood samples of 77 extensively burned patients (>30% of total body surface area) were serially collected in the present study. The expression of HLA-DR on CD14(+) mononuclear cell surface in burned patients were quantified by flow cytometry (using monoclonal antibody, QuantiBRITETM Anti-HLA-DR PE(*)/Anti-Monocyte PerCP-Cy5.5) on days 1, 3, 5, 7, 14, 21 and 28 post burn.
RESULTSThe expressions of HLA-DR on CD14(+) mononuclear cell surface in severely burned patients were significantly lower than those in healthy volunteers from the first day post burn (P < 0.05), and the value of HLA-DR expression was negatively correlated with the burned area (r = -0.7232, P < 0.05). The expression of HLA-DR in patients complicated with multiple organ dysfunction syndrome (MODS) was persistently decreased following major burns, and it was significantly lower than that of non-MODS patients on days 3, 14, 21 and 28 post burn (P < 0.05). The incidence rate of MODS rose markedly along with the lowering of HLA-DR expression, accompanied with poorer prognosis.
CONCLUSIONSExtensive burns could result in marked damage in expression of HLA-DR on CD14(+) mononuclear cell surface and immunologic dysfunction. Quantitative measurement of HLA-DR expression might be of significance in forecasting the development of MODS and prognosis in extensively burned patients.
Adolescent ; Adult ; Burns ; blood ; complications ; immunology ; Female ; Flow Cytometry ; HLA-DR Antigens ; blood ; Humans ; Lipopolysaccharide Receptors ; blood ; Male ; Middle Aged ; Monocytes ; metabolism ; Multiple Organ Failure ; blood ; etiology ; immunology ; Prognosis
8.Hydrogen sulfide in cartilage and its inhibitory effect on matrix metalloproteinase 13 expression in chondrocytes induced by interlukin-1β
Liping PAN ; Yongping CAO ; Licheng WEN ; Weibing CHAI ; Junbao DU ; Hongfang JIN ; Jia LIU ; Xin YANG ; Zhichao MENG ; Heng LIU ; Yunpeng CUI ; Rui WANG ; Hao WU ; Xingtong ZHOU ; Xiang LI ; Zhuoyang LI ; Maimaitijuma TALATIBAIKE
Journal of Peking University(Health Sciences) 2016;48(2):194-202
Objective:To investigate whether endogenous hydrogen sulfide (H2 S)was involved in the pathogenesis of osteoarthritis (OA)and its underlying mechanism,to detect H2 S and its synthases ex-pression in knee cartilage in patients diagnosed with different severity of OA,and to explore the transcrip-tion and expression of gene MMP-13 in chondrocytes treated with IL-1βor H2S.Methods:Synovial fluids of the in-patients with different severity of OA hospitalized in Peking University First Hospital were collected for measurement of H2 S content using methylene blue assay.Articular cartilages of the patients who underwent knee arthroplasty were collected for the cell culture of relatively normal chondrocytes.The chondrocytes were cultured to the P3 generation and H2 S molecular probes were used for detection of endogenous H2 S generation in the chondrocytes.Immunocytochemistry was used to detect the localization of H2 S synthases including cystathionine β-synthase (CBS),cystathionine-γ-lyase (CSE),and mercap-topyruvate sulfurtransferase (MPST)in OA chondrocytes.Western blot was used to quantify the protein expressions of CSE,MPST,and CBS in cartilage tissues of the patients who were diagnosed with OA and underwent knee arthroplasty.The relatively normal human chondrocytes were cultured to passage 3 and then divided into 4 groups for different treatments:(1 )the normal control group,no reagent was added;(2)the IL-1βgroup,5 μg/L of IL-1βwas added;(3)the IL-1β+H2S group,200 μmol/L of NaHS was added 30 min before adding 5 μg/L of IL-1β;(4)the H2 S group,200 μmol/L of NaHS was added. The transcription and expression of gene MMP-13 in chondrocytes of each group were determined with Real-time PCR and Western blot,respectively.And the total NF-κB p65 and phosphorylated NF-κB p65 in chondrocytes were detected with Western blot.Results:The content of H2 S in the synovial fluid of degenerative knee was (14.3 ±3.3)μmol/L.Expressions of endogenous H2 S and its synthases including CBS,CSE and MPST were present in the cytoplasm of chondrocytes.CSE protein expression in Grade 3 (defined by outerbridge grading)cartilage tissues was significantly increased as compared with that of Grade 1 cartilage tissues (1.67 ±0.09 vs.1.26 ±0.11,P<0.05).However,no significant difference of CBS or MPST expression among the different groups was observed.The expression of MMP-13 protein in the IL-1βgroup was significantly higher than that in the normal chondrocytes (1 .87 ±0.67 vs.0.22 ± 0.10,P<0.05 ),and that in the IL-1β+H2 S group was significantly decreased than that in the IL-1βgroup (0.55 ±0.11 vs.1.87 ±0.67,P<0.05),and that in the H2S group had no significant difference compared with that in the normal control group.The transcription of MMP-13 protein in the IL-1βgroup was significantly higher than that in the normal chondrocytes (31.40 ±0.31 vs.1.00 ±0.00,P<0.05), and that in the IL-1β+H2 S group was significantly decreased than that in the IL-1βgroup (24.41 ± 1.28 vs.31.40 ±0.31,P<0.05),and that in the H2S group had no significant difference compared with that in the normal control group.The total NF-κB p65 in the IL-1βgroup was significantly higher than that in the normal chondrocytes (2.13 ±0.08 vs.0.73 ±0.08,P<0.05),and that in the IL-1β+H2S group was significantly decreased than that in the IL-1βgroup (1 .24 ±0.13 vs.2.13 ±0.08,P<0.05 ),and that in the H2 S group had no significant difference compared with that in the normal control group.The phosphorylated NF-κB p65 in IL-1βgroup was significantly higher than that in the normal chondrocytes (1.30 ±0.13 vs.0.19 ±0.04,P<0.05),and that in IL-1β+H2S group was significantly decreased than that in the IL-1βgroup (0.92 ±0.26 vs.1.30 ±0.13,P<0.05),and that in the H2S group had no significant difference compared with that in the normal control group.Conclusion:H2 S affected the cartilage degeneration by partly inhibiting the degradation of extracellular matrix.
9.Construction, expression and bioactivity characterization of targeting toxin DT-VEGF.
Ying-Shuang CHAI ; Xin CHENG ; Li-Hong YAO ; Ai-Jun CHEN ; Hong YU ; Xin-Rui YAN ; Run-Qing JIA ; Guo-Jin HUANG ; Zhi-Qing ZHANG
Chinese Journal of Biotechnology 2004;20(2):192-196
Tumor rapid growth depends on neovascularization. Vascular endothelial growth factor, the main mediator during the occurrence and formation of vascularization, has specific receptors whose expression rate shows difference of orders of magnitude between tumor and the normal tissue, so it can be used to transport toxin molecules to the proliferative tumor endothelial and kill cancer cells. In our experiment, we constructed fusion protein DT-VEGF by linking diphtheria toxin's forward 389 amino acids's gene and VEGF165 via a linker. DT-VEGF is expressed in E. coli and purified. Our experiment proves in can kill vascular endothelial cells specifically, and the inhibition of neovascularization of chicken chorionic membrane is also confirmed.
Angiogenesis Inhibitors
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biosynthesis
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Diphtheria Toxin
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biosynthesis
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genetics
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Escherichia coli
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genetics
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metabolism
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Genetic Vectors
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Humans
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Immunotoxins
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genetics
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Recombinant Fusion Proteins
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biosynthesis
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genetics
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Vascular Endothelial Growth Factors
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biosynthesis
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genetics
10.Study on the relationship between the human leucocyte antigen-DR expression on CD14+ monocytes and sepsis.
Hong-ming YANG ; Yan YU ; Jia-ke CHAI ; Sen HU ; Shu-hong LEI ; Dong-qing ZHANG ; Bai-jiang TAO ; Guo-an ZHANG ; Li-xin HE ; Yong-ming YAO
Chinese Journal of Burns 2007;23(4):272-275
OBJECTIVETo investigate the changes in the expression of HLA-DR on CD14+ monocytes of burn patients with delayed resuscitation, and to analyze the relationship between it and sepsis.
METHODSTwenty-five patients with total burn surface area over 30% TBSA and delayed resuscitation were enrolled in the study, among which 7 were complicated by sepsis during hospitalization. Peripheral blood was collected on 1, 3, 7, 14 and 28 post-burn days (PBD), and the blood of the patients with sepsis were also collected on the 1 and 2 days after the occurrence of sepsis. Twenty healthy volunteers were enrolled as controls. Expression rate of HLA-DR on CD14+ monocytes was determined by flow cytometry. The level of TNF-alpha and IL-10 were measured by ELISA.
RESULTSExpression rate of HLA-DR antigen on CD14+ monocytes in burn patients without sepsis on 1, 3, 7, 14, 28 PBD were (15 +/- 6)%, (7 +/- 5)%, (26 +/- 17)%, (28 +/- 16)% and (47 +/- 16)%, respectively, which were obviously lower than that of healthy people [(92 +/- 10)%, P < 0.01], and it was also markedly lower on 1 and 2 days after the occurrence of sepsis than that of controls and those of patients without sepsis on 1, 7, 14, 28 PBD (P < 0.01). The positive rate and concentration of TNF-alpha in patients with sepsis were obviously higher than that of healthy people and patients without sepsis (P < 0.05 or P < 0.01). There was a negative correlation between the expression rate of HLA-DR on CD14+ monocytes and IL-10 levels, and it showed significant difference on 1, 7, and 28 PBD (r = -0.9963, -0.7459, -0.8474, respectively, P < 0.01).
CONCLUSIONImmune function is suppressed and proinflammatory mediators are excessively released in severely burn patients after delayed resuscitation, especially when complicated with sepsis. Expression of HLA-DR on CD14+ monocytes may be an useful parameter for monitoring the immune function of burn patients.
Burns ; immunology ; metabolism ; HLA-DR Antigens ; metabolism ; Humans ; Interleukin-10 ; blood ; Lipopolysaccharide Receptors ; metabolism ; Monocytes ; immunology ; metabolism ; Sepsis ; immunology ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism