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

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

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 ; Codon, Nonsense ; Factor IX ; genetics ; Genetic Vectors ; Hemophilia B ; genetics ; Humans ; Recombinant Proteins ; genetics

Objective:To explore whether peripheral blood CD8 +T lymphocyte dysfunction is correlated with the programmed death receptor-1 (PD-1) expression in patients with acute-on-chronic liver failure (HBV-ACLF). Methods:Peripheral blood mononuclear cells (PBMC) were collected from patients with HBV-ACLF and healthy controls. CD8 +T lymphocytes number and PD-1 expression condition in CD8 +T lymphocytes were detected by flow cytometry. CD8 +T lymphocytes isolated from peripheral blood of HBV-ACLF patients were further cultured in vitro. One group was added with PD-L1-IgG fusion protein (ACLF+PD-1 group), and the other group was added with IgG fusion protein (ACLF group). Proliferation ability (ki67), cell viability (CD69), and secretion ability of effector cytokines (IL-2, IFN-γ, TNF-α) were analyzed. Results:30 cases with HBV-ACLF and healthy controls were enrolled. CD8 +T lymphocytes absolute number was significantly lower in the peripheral blood of patients with ACLF group (333.88 ± 147.74)/μl than healthy controls (872.50 ± 206.64)/μl ( P < 0.001). PD-1 expression in peripheral blood CD8 +T lymphocytes were significantly increased in ACLF group (13.33% ± 2.52%), ( P = 0.027) than healthy controls (7.02% ± 2.12%). In in vitro culture, compared with healthy controls, the peripheral blood CD8 +T lymphocytes cell viability (CD69), proliferation ability (ki67) (all P ??< 0.001), and the level of cytokine production (IL-2, IFN-γ, TNF-α) (all P < 0.05) were equally weakened in patients with ACLF group. Compared with ACLF group, CD8 +T cell viability (CD69), proliferation ability (KI67) (all P < 0.05), and the level of cytokine production were weakened in ACLF+PD-1 group (all P < 0.05). Conclusion:HBV-ACLF patients have CD8 +T lymphocyte dysfunction. Therefore, PD-1 may have correlation in the regulation of CD8 +T lymphocyte dysfunction in ACLF patients.

Background and Objective：Glutathione S-transferase M1 (GSTM 1)deficiency may increase the risk of nasopharyngeal carcinoma (NPC).This study was to evaluate the correlation of the single nucleotide polymorphism (SNP) in the coding region of GSTMl gene to NPC susceptibility in southern China population.Methods：In total 239 NPC patients and 286 age-matched healthy controls were entered into the study.Among them.225 out of 239 NPC patients and 273 out of 286 controls were used for statisticaI analysis.SNP screening of all exons.relevant intron-exon boundaries.and the promoter region of GSTM 1，in total 4739bp，was performed by PCR direcl sequencing.The loci T1270533G and C1256088Cwere selected for the case-controI study using the tetra-Primer ARMS-PCR.as well as the sequencing method.Results：In totaf 29 SNPs of GSTM 1 were identified by sequencing.Missense mutation occurred in the polymorphic loci of T1270533G and C1256088C.However.no evident relationships between the variant of T1270533G and clinicaI phenotypes of NPC were obsewed in the NPC group and healthy control group(OR=0.1 70，95%CI=0.95-0.306for homozygote TT).The deletion frequency of C1256088C was 45%(45/100)for NPC patients and 42%(42/100)for controls.Conclusions：The polymorphism of T1270533G does not affect the detoxification function of GSTM1.The T1270533G JOCUS has no apparent association with genetic susceptibility to NPC in the southern China population.The IOSS rate of C1256088C is high in this study.

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

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.

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

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

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