Analysis of Pharmacological Activities and Mechanisms of Essential Oil in Leaves of C. grandis 'Tomentosa' by GC-MS/MS and Network Pharmacology.

BACKGROUND: Citrus grandis 'Tomentosa,' a fruit epicarp of C. grandis 'Tomentosa' or C. grandis (L.) Osbeck is widely used in health food and medicine. Based on our survey results, there are also rich essential oils with bioactivities in leaves, but the chemical compounds in this part and relevant pharmacological activities have never been studied systematically. Therefore, this study was to preliminarily decipher the pharmacological activities and mechanisms of the essential oil in leaves of C. grandis 'Tomentosa' by an integrated network pharmacology approach. METHODS: Essential oil compositions from leaves ofC. grandis 'Tomentosa' were identified using GC-MS/MS. And then, the targets of these oil compositions were predicted and screened from TCMSP, SwissTargetPrediction, STITCH and SEA databases. STRING database was used to construct the protein-protein interaction networks, and the eligible protein targets were input into WebGestalt 2019 to carry out GO enrichment and KEGG pathway enrichment analysis. Based on the potential targets, disease enrichment information was obtained by TTD databases. Cytoscape software was used to construct the component-target-disease network diagrams. RESULTS: Finally, 61 essential oil chemical components were identified by GC-MS/MS, which correspond to 679 potential targets. Biological function analysis showed 12, 19, and 12 GO entries related to biological processes, cell components and molecular functions, respectively. 43 KEGG pathways were identified, of which the most significant categories were terpenoid backbone biosynthesis, TNF signaling pathway and leishmaniasis. The component-target-disease network diagram revealed that the essential oil compositions in leaves of C. grandis 'Tomentosa' could treat tumors, immune diseases, neurodegenerative diseases and respiratory diseases, which were highly related to CHRM1, PTGS2, CASP3, MAP2K1 and CDC25B. CONCLUSION: This study may provide new insight into C. grandis 'Tomentosa' or C. grandis (L.) Osbeck and may provide useful information for future utilization and development.

Downregulation of lncRNA NEAT1 Ameliorates LPS-Induced Inflammatory Responses by Promoting Macrophage M2 Polarization via miR-125a-5p/TRAF6/TAK1 Axis.

The lncRNA nuclear enriched abundant transcript 1 (NEAT1) promotes sepsis-inflammatory responses and acute kidney injury (AKI), but little known about the underlying mechanisms. This study aims to investigate the roles of NEAT1 in regulating macrophage polarization and its potential for alleviating inflammatory responses during sepsis pathogenesis. Mouse RAW264.7 macrophages were treated with lipopolysaccharide (LPS) as a cellular inflammatory model. NEAT1 shRNA, miR-125a-5p mimics, and TRAF6-overexpressing vector were used to transfect RAW264.7 cells. NEAT1, miR-125a-5p, and mRNA levels of functional genes were detected by quantitative RT-PCR. Protein abundances were analyzed by western blotting. Macrophage polarization was evaluated by flow cytometry. The bindings of miR-125a-5p with NEAT1 or TRAF6 gene were validated by dual luciferase reporter assay. LPS treatment promoted NEAT1 and suppressed miR-125a-5p expression in mouse macrophage cells. NEAT1 silencing by shRNAs promoted macrophage M2 polarization under LPS treatment, which upregulated miR-125a-5p expression, repressed TRAF6 expression and TAK1 protein phosphorylation in macrophages. These cellular and molecular changes induced by NEAT1 shRNAs were abrogated by miR-125a-5p inhibitors. Moreover, miR-125a-5p mimics suppressed TRAF6 expression and TAK1 protein phosphorylation in LPS-treated macrophages, thus causing macrophage M2 polarization under LPS treatment. TRAF6 overexpression abrogated the miR-125a-5p mimics-induced macrophage M2 polarization. miR-125a-5p could directly bind to NEAT1 or TRAF6 gene in macrophages. lncRNA NEAT1 knockdown ameliorates LPS-induced inflammation by promoting macrophage M2 polarization via miR-125a-5p/TRAF6/TAK1 axis.

Alpha-Lipoic Acid Suppresses Extracellular Histone-Induced Release of the Infammatory Mediator Tumor Necrosis Factor-α by Macrophages.

BACKGROUND/AIMS: This study investigated signaling pathways via which extracellular histones induce the pro-inflammatory cytokine tumor necrosis factor-α (TNF-α) release from the macrophage cell line RAW 264.7 and the anti-inflammatory efficacy of the antioxidant alpha-lipoic acid (ALA). METHODS: ELISA and western blotting analyses were conducted to detect the release of TNF-α from histone-stimulated RAW 264.7 macrophages and the associated phospho-activation of MAPKs (ERK and p38) and NF-κB p65. The effects of ALA on the release of TNF-α and phospho-activation of the MAPKs and NF-κB p65 were studied. P < 0.05 was considered statistically significant. RESULTS: Extracellular histones dose-dependently induced TNF-α release from RAW 264.7 cells and increased the phosphorylation of p38, ERK, and NF-κB p65. TNF-α release was markedly suppressed by p38, ERK, and NF-kB inhibitors. ALA reduced histone-induced TNF-α release, ERK/p38 MAPK activation, and NF-kB activation without affecting macrophage viability. CONCLUSION: Histones induce TNF-α release from macrophages by activating the MAPK and NF-kB signaling pathways, while ALA suppresses this response by inhibiting ERK, p38 and NF-kB. These findings identify potentially critical inflammatory signaling pathways in sepsis and molecular targets for sepsis treatment.

Procalcitonin levels in patients with positive blood culture, positive body fluid culture, sepsis, and severe sepsis: a cross-sectional study.

BACKGROUND: Numerous investigations on procalcitonin (PCT) have been carried out, although few with large sample size. To deal with the complexity of sepsis, an understanding of PCT in heterogeneous clinical conditions is required. METHODS: Hospitalized patients aged 10-79 years were included in this retrospective and cross-sectional study. PCT tests were assayed within 2 days of blood culture. RESULTS: A total of 2952 cases (from 2538 patients) were enrolled in this study, including 440 cases in the 'positive BC' group, 123 cases in the 'positive body fluid culture' group, and 2389 cases in the 'negative all culture' group. Median PCT values were 4.53 ng/ml, 2.95 ng/ml, and 0.49 ng/ml, respectively. Median PCT values in the gram-negative BC group and gram-positive BC group, respectively, were 6.99 ng/ml and 2.96 ng/ml. Median PCT values in the 'positive hydrothorax culture' group, 'positive ascites culture' group, 'positive bile culture' group, and 'positive cerebrospinal fluid culture' group, respectively, were 1.39 ng/ml, 8.32 ng/ml, 5.98 ng/ml, and 0.46 ng/ml. In all, 357 cases were classified into the 'sepsis' group, 150 of them were classified into the 'severe sepsis' group. Median PCT values were 5.63 ng/ml and 11.06 ng/ml, respectively. CONCLUSIONS: PCT could be used in clinical algorithms to diagnose positive infections and sepsis. Different PCT levels could be related to different kinds of microbemia, different infection sites, and differing severity of sepsis.

[Assessment of the value of acute physiology and chronic health evaluation II/IV prognostic models in elderly patients with sepsis].

OBJECTIVE: To assess and compare the performance of acute physiology and chronic health evaluation II/IV (APACHEII/IV) prognostic models in elderly patients with sepsis. METHODS: A totally of 82 elderly patients with sepsis were retrospectively assessed in geriatric intensive care unit of General Hospital of Guangzhou Military Command between July 2011 and December 2011. APACHEII/IV scores were recorded within 24 hours after admission. The prognosis accuracy of both scores was assessed by area under the receiver operating characteristic curve (AUC). Based on the best cutoff value corresponding with the highest accuracy, patients were divided into the low and high risk of hospital mortality group. The predictive power of APACHEII/IV in total population and subgroups was compared. RESULTS: Patients with severe sepsis constituted 57.3% (47/82) of all patients with sepsis, and hospital mortality was 61.0% (50/82). APACHEII/IV scores of the patients were 17.5±6.3 and 55.8±22.3, and mortality rate was 22.5% (18.4/82) and 17.9% (14.6/82) respectively, with significant differences compared with actual mortality (both P<0.01). Both APACHEII/IV scores showed underestimation of hospital mortality in total population [standardized mortality ratio (SMR) with APACHEII=2.71, 95% confidence interval (95%CI) 1.92-3.48 and SMR with APACHEIV=3.33, 95%CI 2.79-4.37]. APACHEII (AUC 0.664±0.066), and APACHEIV presented poor estimation(AUC 0.716±0.056). There was no difference in accuracy in prognosticating hospital death prognosis between the two APACHE models (Z=0.991, P=0.322). Cutoff values of APACHEII/IV were >11 and >59. According to the value, patients were divided into the low and high risk hospital mortality group. There was no significant difference between actual mortality and prognostic mortality in APACHEII low risk group [0-11, 20.0% (3/15) vs. 1.6% (0.2/15), Z=-1.023, P=0.306]. The actual mortality in high risk group with APACHEII (>11) was significantly higher than prognostic mortality [70.1% (47/67) vs. 27.2% (18.2/67), t=6.989, P=0.000]. In the high risk group, APACHEII underestimated mortality (SMR=2.58, 95%CI 2.22-3.51). The actual mortality of the low (0-59) and high (>59) risk group of APACHEIV were higher than prognostic mortality [lower risk group: 44.0% (22/50) vs. 7.5% (3.8/50), Z=-2.235, P=0.025; higher risk group: 87.5% (28/32) vs. 34.1% (10.9/32), Z=-4.712, P=0.000]. Two groups of patients with APACHEIV score, the mortality was underestimated (low risk group: SMR=5.90, 95%CI 5.19-7.07; high risk group: SMR=2.56, 95%CI 2.07-3.24). Mortality rate of the low risk group with APACHEIV score was prone to be underestimated. CONCLUSIONS: The accuracy of APACHEII/IV are not ideal in foretelling mortality rate. Hospital mortality was underestimated with APACHEII in high risk patients, and it was underestimated with APACHEIV both in low and high risk patients, and it is even more prone to be underestimated in low risk group of APACHEIV. More accurate prognostic modality is in need in elderly patients with sepsis.

Histones-mediated lymphocyte apoptosis during sepsis is dependent on p38 phosphorylation and mitochondrial permeability transition.

Lymphocyte apoptosis is one reason for immunoparalysis seen in sepsis, although the triggers are unknown. We hypothesized that molecules in plasma, which are up-regulated during sepsis, may be responsible for this. In this study, peripheral lymphocyte apoptosis caused by extracellular histones was confirmed both in mouse and human primary lymphocytes, in which histones induced lymphocyte apoptosis dose-dependently and time-dependently. To identify which intracellular signal pathways were activated, phosphorylation of various mitogen-activated protein kinases (MAPKs) were evaluated during this process, and p38 inhibitor (SB203580) was used to confirm the role of p38 in lymphocyte apoptosis induced by histones. To investigate the mitochondrial injury during these processes, we analyzed Bcl2 degradation and Rhodamine 123 to assess mitochondrial-membrane stability, via cyclosporin A as an inhibitor for mitochondrial permeability transition (MPT). Then, caspase 3 activation was also checked by western-blotting. We found that p38 phosphorylation, mitochondrial injury and caspase 3 activation occurred dose-dependently in histones-mediated lymphocyte apoptosis. We also observed that p38 inhibitor SB203580 decreased lymphocyte apoptotic ratio by 49% (P<0.05), and inhibition of MPT protected lymphocytes from apoptosis. Furthermore, to investigate whether histones are responsible for lymphocyte apoptosis, various concentrations of histone H4 neutralization antibodies were co-cultured with human primary lymphocytes and plasma from cecal ligation and puncture (CLP) mice or sham mice. The results showed that H4 neutralization antibody dose-dependently blocked lymphocyte apoptosis caused by septic plasma in vitro. These data demonstrate for the first time that extracellular histones, especially H4, play a vital role in lymphocyte apoptosis during sepsis which is dependent on p38 phosphorylation and mitochondrial permeability transition. Neutralizing H4 can inhibit lymphocyte apoptosis, indicating that it could be a potential target in clinical interventions for sepsis associated immunoparalysis.

Intraoperative microvascular Doppler monitoring in intracranial aneurysm surgery / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Surgical treatment of intracranial aneurysms is often compromised by incomplete exclusion of the aneurysm or stenosis of parent vessels. Intraoperative microvascular Doppler (IMD) is an attractive, noninvasive, and inexpensive tool. The present study aimed to evaluate the usefulness and reliability of IMD for guiding clip placement in aneurysm surgery.</p><p><b>METHODS</b>A total of 92 patients with 101 intracranial aneurysms were included in the study. IMD with a 1.5-mm diameter, 20-MHz microprobe was used before and after clip application to confirm aneurysm obliteration and patency of parent vessels and branching arteries. IMD findings were verified postoperatively with digital subtraction angiography (DSA) or dual energy computed tomography angiography (DE-CTA). Ninety consecutive patients, harboring 108 aneurysms, who underwent surgery without IMD was considered as the control group.</p><p><b>RESULTS</b>The microprobe detected all vessels of the Circle of Willis and their major branches. Clips were repositioned in 24 (23.8%) aneurysms on the basis of the IMD findings consistent with incomplete exclusion and/or stenosis. IMD identified persistent weak blood flow through the aneurismal sac of 11 of the 101 (10.9%) aneurysms requiring clip adjustment. Stenosis or occlusion of the parent or branching arteries as indicated by IMD necessitated immediate clip adjustment in 19 aneurysms (18.8%). The mean duration of the IMD procedure was 4.8 minutes. The frequency of clip adjustment (mean: 1.8 times per case) was associated with the size and location of the aneurysm. There were no complications related to the use of IMD, and postoperative angiograms confirmed complete aneurysm exclusion and parent vessel patency. About 8.3% (9/108) aneurysms were unexpectedly incompletely occluded, and 10.2% (11/108) aneurysms and parent vessel stenosis without IMD were detected by postoperative DSA or DE-CTA. IMD could reduce the rate of residual aneurysm and unanticipated vessel stenosis which demonstrated statistically significant advantages compared with aneurysm surgery without IMD.</p><p><b>CONCLUSION</b>IMD is a safe, easily performed, reliable, and valuable tool that is suitable for routine use in intracranial surgery, especially in complicated, large, and giant aneurysms with wide neck or without neck.</p>

Intracerebroventricular transplantation of human amniotic epithelial cells ameliorates spatial memory deficit in the doubly transgenic mice coexpressing APPswe and PS1ΔE9-deleted genes / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Human amniotic epithelial cells (HAECs), which have characteristics of both embryonic and pluripotent stem cells, are therefore a candidate in cell therapy without creating legal or ethical problems. In the present study, we aimed to investigate the effects of intracerebroventricular transplantation of HAECs on doubly transgenic mice of Alzheimer's disease (AD) coexpressing presenilin-1 (PS1) and mutant Sweden amyloid precursor protein (APPswe) genes.</p><p><b>METHODS</b>The offspring mice genotypes were detected using PCR identification of APPswe and PS1 gene. The doubly transgenic (TG) mice (n = 20) and wild-type (WT) mice (n = 20) were randomly divided into two groups respectively: the transplantation group treated with HAECs and the control group with phosphate buffered saline. Six radial arm water maze test was used to assess the spatial memory in the TG and WT mice. Amyloid plaques and neurofibrillary tangles were analyzed using congo red and acid-silver methenamine staining respectively. Immunofluorescence cytochemistry was used to track the survival of HAECs. Immunohistochemistry was used to determine the expression of octamer-binding protein 4 (Oct-4) and Nanog in the HAECs. High performance liquid chromatography was used to measure acetylcholine in hippocampus. The density of cholinergic neurons in basal forebrain and nerve fibers in hippocampus was measured using acetylcholinesterase staining.</p><p><b>RESULTS</b>Amyloid deposition occurred in hippocampus and frontal cortex in the double TG mice aged 8 months, but not in WT mice. The results also showed that transplanted HAECs can survive for at least 8 weeks and migrate to the third ventricle without immune rejection. The graft HAECs can also express the specific marker Oct-4 and Nanog of stem cell. Compared with the control group, transplantation of HAECs can not only significantly improve the spatial memory of the TG mice, but also increase acetylcholine concentration and the number of hippocampal cholinergic neurites.</p><p><b>CONCLUSIONS</b>These results demonstrate that intracerebroventricular transplantation of HAECs can improve the spatial memory of the double TG mice. The higher content of acetylcholine in hippocampus released by more survived cholinergic neurites is one of the causes of this improvement.</p>

[Effect of continuous hemofiltration for treatment of multiple organ dysfunction syndrome after acute myocardial infarction in elderly patients].

OBJECTIVE: To study the effect of continuous hemofiltration (CH) on the prognosis of multiple organ dysfunction syndrome (MODS) after acute myocardial infarction (AMI) in elderly patients. METHODS: Thirty-four elderly patients with MODS after AMI admitted to intensive care unit (ICU) were grouped into continuous hemofiltration (CH) group and non-CH (NCH) group. The Acute Physiology and Chronic Health Evaluation (APACHEII) scores and Marshall scores were assessed upon admission in ICU and 7 days after the admission. The mortality rates of the patients within 28 and 90 days after admission to ICU were calculated, and the changes in APACHEII scores and Marshall scores were compared between the two groups. RESULTS: The APACHEII scores and Marshall scores showed no significant difference between the two upon admission to the ICU, but significantly decreased in CH group 7 days after the admission (P<0.05 and P<0.01 respectively). The APACHEII scores increased significantly in NCH group (P<0.01) 7 days after the admission while the Marshall scores remained unchanged (P>0.05). The overall mortality rates at 28 and 90 days were 41.18% and 61.76%, respectively. A significant difference was noted in the mortality rate at 28 days between the two groups (P<0.05), but not in the rate at 90 days (P>0.05). CONCLUSION: CH can improve the organic functions and the short-term outcome of elderly patients with MODS after AMI, but has no positive effect on their long-term outcomes.

Serum and lung endothelin-1 increased in a canine model of ventilator-induced lung injury.

BACKGROUND: Nitric oxide (NO) plays an important role in acute lung injury (ALI), acute respiratory distress syndrome (ARDS), and in ventilator-induced lung injury (VILI). A change in the balance of endothelin-1 (ET-1) and NO in the ALI/ARDS can also add to these problems. However, the profile of ET-1 and the balance of ET-1 and NO are still unknown in a VILI model. METHODS: Models of oleic acid induced ALI were established in dogs; these models were then randomized into three groups undergone different tidal volume (VT) mechanical ventilation, which included a VT6 group (VT equaled to 6 ml/kg body weight, positive end expiratory pressure (PEEP) equaled to 10 cmH2O, n = 6), a VT10 group (VT equaled to 10 ml/kg body weight, PEEP equaled to 10 cmH2O, n = 4) and a VT20 group (VT equaled to 20 ml/kg body weight, PEEP equaled to 10 cmH2O, n = 6) for 6-hour ventilation. The levels of ET-1 and NO in serum and tissue homogenates of lung were observed throughout the trial. RESULTS: PaO2 was increased after mechanical ventilation, but hypercapnia occurred in the VT6 group. The magnitudes of lung injury in the VT20 group were more severe than those in the VT6 group and the VT10 group. Serum levels of ET-1 and NO increased after ALI models were established and slightly decreased after a 6-hour ventilation in both the VT6 group and the VT20 group. The serum ET-1 level in the VT20 group was higher than that in the VT6 group and the VT10 group after the 6-hour ventilation (P < 0.05) while the serum NO levels were similar in the three groups (all P > 0.05). There was no significant difference in serum ratio of ET-1/NO between any two out of three groups (P > 0.05), although there was a significant positive relationship between serum ET-1 and serum NO (r = 0.80, P < 0.01). The levels of ET-1 and NO in the lung were increased after ventilation. The lung ET-1 level in the VT20 group was significantly higher than that in the VT6 group and VT10 group (both P < 0.05) while there was no significant difference in lung NO levels between two groups (P > 0.05). In the lung tissue, the ratio of ET-1/NO was significantly higher in the VT20 group than in the VT6 group and VT10 group after the 6-hour ventilation (P < 0.05) as there was a significant positive relationship between ET-1 and NO in the lung (r = 0.54, P < 0.05). CONCLUSIONS: The production of ET-1 and NO was increased in serum and lung tissue in a VILI model. But the ET-1 levels increased much more than the NO levels in the lung, though there was a significant positive relationship between levels of ET-1 and NO. These results showed that there was an interaction between ET-1 and NO in a VILI model and changing the balance of ET-1 and NO levels might contribute to the pathophysiologic process of VILI.

[Changes of thyroxin and monocyte human leukocyte antigen-DR expression in senior patients with sepsis].

OBJECTIVE: To investigate the changes of thyroxin and monocyte human leukocyte antigen-DR expression in senior patients with sepsis and explore their clinical significance. METHODS: According to the 2001 SCCM/ESICM/ACCP/ATS/SIS international sepsis definitions, 125 senior patients with sepsis free of thyroid conditions were divided into non-severe sepsis group (n=86) and severe sepsis group (n=39), with another 30 healthy subjects as the control. Thyroid function was assayed by chemoluminescence method in these patients and monocyte HLA-DR expression was determined by flow cytometry. RESULTS: Compared with the control group and non-severe sepsis cases, the levels of free T3 (FT3), free T4 (FT4), T3, T4 and monocyte HLA-DR expression were significantly lower in severe sepsis cases (P<0.05), but the levels of thyroid stimulating hormone (TSH) were comparable between the 3 groups (P>0.05). The non-severe sepsis cases showed significantly lower levels of FT3, FT4, T3, T4, TSH and monocyte HLA-DR expression than the control group (P<0.05). In severe sepsis group, the levels of FT3, FT4, T3, T4 and monocyte HLA-DR expression showed significant differences between the fatal cases and surviving cases (P<0.05). CONCLUSION: The levels of thyroxin and monocyte human leukocyte antigen-DR expression are obviously lower in senior patients with severe sepsis, and their detection may well indicate the severity of the condition and help make prognostic judgment.

[Role of assessment and monitoring of human leucocyte antigen-DR on CD14+ monocyte in postoperative infection in patients after orthotopic liver transplantation].

OBJECTIVE: To investigate the changes in expression level of human leucocyte antigen-DR (HLA-DR) on CD14(+) monocyte (CD14(+)/HLA-DR) in the patients after orthotopic liver transplantation, and its role in monitoring postoperative infection. METHODS: Sixty-three patients with liver transplantation were divided into three groups, non-infection group with 47 cases, infection group with 10 cases and septic shock group with 6 cases [according to the definition of septic shock of American College of Chest Physicians/Society for Critical Care Medicine (ACCP/SCCM)]. CD14(+)/HLA-DR expression ratio was assessed with flow cytometer, and its clinical implication was evaluated by receiver operating characteristic (ROC) curve assay. RESULTS: CD14(+)/HLA-DR expression ratio in infection group [(29.6+/-7.2)%] and septic shock group [(16.3+/-10.5)%] were significantly lower than that in non-infection group [(62.3+/-18.3)%, both P<0.01], but no significant difference of CD14(+)/HLA-DR expression ratio was found between infection group and septic shock group (P=0.128). Total area under ROC curve of CD14(+)/HLA-DR expression ratio for the infection was 0.965, its sensitivity and specificity at 36.35% cut off were 100.0% and 93.6%, respectively. Total area under ROC curve of CD14(+)/HLA-DR expression ratio to predict septic shock was 0.968, its sensitivity and specificity at 31.97% cut off were 100.0% and 87.7%, respectively. Comparing the change of CD14(+)/HLA-DR expression, it was lower in the infection group and septic shock group (P<0.05 and P<0.01), and the expression rate was lowest during period of serious infection in the two groups [infection group: (29.6+/-7.2)%, septic shock group: (16.3+/-0.5)%, all P<0.01]. CONCLUSION: For the patients with possible infection after liver transplantation, sequential assessment of CD14(+)/HLA-DR expression ratio would be a good marker for the judgment of patient's conditions and outcome. CD14(+)/HLA-DR expression ratio below 36.35% could be used as the prewarning value for the diagnosis of postoperative infection, and 31.97% could be used as the critical value for the diagnosis of septic shock.

[Early bronchoscopic sputum suction in elderly patients with aspiration pneumonia-induced acute heart and lung failure].

OBJECTIVE: To observe the effect of early bronchoscopic sputum suction in elderly patients with acute heart and lung failure due to aspiration pneumonia. METHODS: Comprehensive treatments were administered for 52 elderly patients with acute heart and lung failure resulting from aspiration pneumonia, and in 27 of the patients, bronchoscopic sputum suction was performed with the other 25 serving as the control group. The indices of the heart and lung functions (central venous pressure, left ventricular ejection fraction, arterial blood partial pressures of oxygen and carbon dioxide) were measured after the treatment and compared between the two groups. RESULTS: Patients receiving bronchoscopic suction showed faster recovery of normal central venous pressure and left ventricular ejection fraction and more rapid increment of arterial partial pressure of oxygen and reduction of carbon dioxide partial pressure than those without the suction (P<0.01). CONCLUSION: Early bronchoscopic sputum suction can be one of the effective emergency measures for rescuing acute heart and lung failure due to aspiration pneumonia.

[Therapeutic effect of low-dose thyroxin in elderly patients with refractory heart failure and euthyroid sick syndrome].

OBJECTIVE: To investigate the therapeutic effect of low-dose thyroxin in elderly patients with refractory heart failure (RHF) and euthyroid sick syndrome (ESS). METHODS: Fifty-four elderly patients with RHF and ESS were randomized into conventional treatment group (n=32) and L-thyroxine group with additional oral L-thyroxine at the daily dose of 6.25-25 microg (n=22). The changes in the plasma levels of brain natriuretic peptides (BNP), left ventricular ejection fraction (LVEF), and cardiac function (NYHA level) of the two groups were compared after 1 month of treatment. RESULTS: Five patients receiving conventional treatment died due to severe arrhythmia during the treatment, and in the other 27 patients, the levels of plasma BNP, LVEF, and cardiac function showed no significant improvements after 1 month of treatment (P>0.01). In L-thyroxine group, no death or severe arrhythmia occurred, and the levels of plasma BNP, LVEF, and cardiac function were significantly improved after the treatment (P<0.01). No thyrotoxicosis occurred during the administration of L-thyroxine in the latter group. CONCLUSION: Low-dose L-thyroxine in addition to the conventional treatments may enhance the therapeutic effect in elderly patients with RHF and ESS.

[Effect of C5a on expression of thrombomodulin in endothelial cells in vitro].

OBJECTIVE: To investigate the effect of C5a on the expression of thrombomodulin (TM) in human umbilical vein endothelial cells (HUVECs). METHODS: HUVECs cultured in vitro were stimulated with C5a for 8, 12, 16, 20 hours in a concentration of 200 microg/L, and also with different concentrations of 100, 200, 300 microg/L for 12 hours respectively. The levels of both mRNA and protein expression of TM was detected by real-time polymerase chain reaction (PCR) and Western blotting respectively, and the dose and time dependent effects of C5a on the expression of TM were evaluated. RESULTS: C5a down-regulated the TM expression at both protein and mRNA level. The down-regulation was time-dependent [(93.11+/- 1.57) x10(-2) , (71.05+/-3.39)x10(-2), (65.48+/-4.28)x10(-2), (62.69+/-4.03)x10(-2)at protein level and (301.71+/-80.40)x10(-6), (38.29+/-20.24)x10(-6), (8.82+/-2.66)x10(-6), (7.05+/-0.80)x10(-6) at mRNA level, all P<0.05] and dose-dependent [(113.25+/-3.97)x10(-2), (80.18+/-2.56)x10(-2), (73.22+/- 4.36)x10(-2) at protein level and (401.77+/-20.46)x10(-6), (31.12+/-3.51)x10(-6), (18.19+/-1.46)x10(-6) at mRNA level , all P<0.05]. When concentration of C5a at 300 microg/L was used to stimulate HUVECs for longer than 12 hours, the lowering of TM at protein level was slowed down obviously. And concentration of C5a at 200 microg/L was used to stimulate HUVECs for 12 hours, the lowering of TM at mRNA level was slowed down obviously. CONCLUSION: C5a can depress the gene expression of TM, and then affect the protein's translation. By this means, C5a can lead to hypercoagulability and inflammatory injuries in sepsis.

[Effect of mechanical ventilation with different tidal volume on small intestine in dogs suffering from acute lung injury].

OBJECTIVE: To compare the effect of different tidal volume (VT) on intestinal tissue in oleic acid-induced acute lung injury (ALI) dogs undergoing mechanical ventilation (MV). METHODS: ALI was induced with oleic acid in dogs. While all of them were undergoing MV, they were randomized into two groups: low VT group (n=6), with VT 6 ml/kg, positive end-expiratory pressure (PEEP) 10 cm H2O (1 cm H2O= 0.098 kPa), and large VT group (n=6), with VT 20 ml/kg, PEEP 10 cm H2O. MV with different VT was maintained for 6 hours. After 6 hours, dogs were sacrificed by exsanguination. Pathological changes in small bowel tissues were observed with hematoxylin and eosin (HE) staining, and cellular apoptosis detected with terminal deoxynucleotidyl transferase-mediated dUTP biotin nick end labeling (TUNEL) in situ apoptosis determination. RESULTS: After 6-hour MV, the degree of flatulence was severe in the large VT group, but no flatulence was observed in the low VT group. Bowel injury score was lower in the low VT group than in large VT group [(3.17+/-0.75) scores vs. (2.00+/-0.89) scores]. There was significant difference between two groups (P<0.01). Apoptosis-positive cells were rare in the small bowel tissues in both groups. CONCLUSION: MV with large VT can induce bowel dysfunction, in contrast low VT MV does not produce small bowel dysfunction.

[Monocyte human leukocyte antigen-DR expression in elderly patients with sepsis].

OBJECTIVE: To investigate the changes of human leukocyte antigen DR (HLA-DR) expression in the monocytes of elderly patients with sepsis. METHODS: A total of 213 elderly patients were divided into 4 groups according to the criteria defined by the American College of Chest Physicians/ Society of Critical Care Medicine (ACCP/SCCM), namely the non-sepsis group (group A), sepsis group (group B), severe sepsis group (group C) and septic shock group (group D). Another 60 healthy subjects were recruited as the control group. HLA-DR expression in the monocytes from each subject was determined by flow cytometry, and the HLA-DR levels were compared between the groups. RESULTS: Compared with that of the healthy control, HLA-DR expression was significantly increased in group A and lowered in group D. No significant differences were found between the control group and groups B and C. HLA-DR expression was significantly different between groups A, B, C, and D (P<0.01), increasing in the order of group D (32.74-/+13.4)%, group C (61.9-/+14.29)%, group B (69.99-/+12.8)%, and group A (85.06-/+15.37)%. CONCLUSIONS: Although with multiple organ diseases and repeated or long-term hospitalization, the non-septic elderly patients do not show lowered expression of HLA-DR. In the elderly patients with sepsis and severe sepsis, HLA-DR expression is not a sensitive index, but obviously lowered HLA-DR expression may serve as a specific indicator in elderly patients with septic shock.

[Clinical value of the model for end-stage liver disease score in predicting the prognosis of liver transplantation in patients with end-stage liver disease].

OBJECTIVE: To assess the value of the model for end-stage liver disease (MELD) in predicting the early-stage outcome of liver transplantation in patients with end-stage liver disease. METHODS: The MELD scores of 87 liver transplantation recipients with end-stage liver disease were calculated, and their early-stage complications and mortality were analyzed. RESULTS: The incidence of severe complications was 20.7%; in these recipients, with the 28-day and 3-month survival rates of 89.7%; and 88.5%;, respectively. The mean MELD scores showed significant differences between the complication-free group and survival group (14.6 vs 12.9, P<0.05), and also between the complication group and death group (21.6 vs 29.4, P<0.05). Compared to patients with MELD no greater than 15, patients with MELD between 16 and 24 showed significantly increased complication rate but had comparable survival rate (P>0.05); but in patients with MELD no less than 25, the survival rate was significantly decreased with also increased complication rate. CONCLUSIONS: A higher MELD score before liver transplantation is associated with greater likeliness of early-stage complication rate and mortality. High MELD score (over 25) can be a useful index in predicting severe complications and death in patients undergoing liver transplantation.

A novel full-length gene of human ribosomal protein L14.22 related to human glioma / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>This study was undertaken to obtain differentially expressed genes related to human glioma by cDNA microarray and the characterization of a novel full-length gene.</p><p><b>METHODS</b>Total RNA was extracted form human glioma and normal brain tissue, and mRNA was used as a probe. The results of hybridization procedure were scanned with the computer system. The gene named 507E08 cone was subsequently analyzed by northern blot, bioinformatic approach, and protein expression.</p><p><b>RESULTS</b>Fifteen differentially expressed genes were obtained from human glioma by hybridization and scanning for four times. Northern blot analysis confirmed that the 507E08 clone was low expressed in human brain tissue and over expressed in human glioma tissues. The analysis of BLASTn and BLASTx showed that the 507E08 clone was a novel full-length gene, which codes 203 amino acid of protein and is called human ribosomal protein 14.22 gene. The nucleotide sequence had been submitted to the GenBank with the accession number of AF329277. After expression in E. coli., protein yielded a major band of apparent molecular mass 22 kDa on an SDS-PAGE gel.</p><p><b>CONCLUSIONS</b>cDNA microarray technology can be successfully used to identify differentially expressed genes. The novel full-length gene of human ribosomal protein 13.22 may be correlated with the development of human glioma.</p>

Transplantation of human amniotic epithelial cells improves hindlimb function in rats with spinal cord injury / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Human amniotic epithelial cells (HAECs), which have several characteristics similar to stem cells, therefore could possibly be used in cell therapy without creating legal or ethical problems. In this study, we transplanted HEACs into the injured spinal cord of rats to investigate if the cells can improve the rats' hindlimb motor function.</p><p><b>METHODS</b>HAECs were obtained from a piece of fresh amnion, labeled with Hoechst33342, and transplanted into the site of complete midthoracic spinal transections in adult rats. The rats (n = 21) were randomly divided into three groups: Sham-operation group (n = 7), cells-graft group (n = 7), and PBS group (n = 7). One rat of each group was killed for histological analysis at the second week after the transplantation. The other six rats of each group were killed for histological analysis after an 8-week behavioral testing. Hindlimb motor function was assessed by using the open-field BBB scoring system. Survival rate of the graft cells was observed at second and eighth weeks after the transplantation. We also detected the myelin sheath fibers around the lesions and the size of the axotomized red nucleus. A one-way ANOVA was used to compare the means among the groups. The significance level was set at P < 0.05.</p><p><b>RESULTS</b>The graft HAECs survived for a long time (8 weeks) and integrated into the host spinal cord without immune rejection. Compared with the control group, HAECs can promote the regeneration and sprouting of the axons, improve the hindlimb motor function of the rats (BBB score: cells-graft group 9.0 +/- 0.89 vs PBS group 3.7 +/- 1.03, P < 0.01), and inhibit the atrophy of axotomized red nucleus [cells-graft group (526.47 +/- 148.42) microm(2) vs PBS group (473.69 +/- 164.73) microm(2), P < 0.01].</p><p><b>CONCLUSION</b>Transplantation of HAECs can improve the hindlimb motor function of rats with spinal cord injury.</p>