An Immune Risk Score Predicts Survival of Patients with Acute Myeloid Leukemia Receiving Chemotherapy.

PURPOSE: Prediction models for acute myeloid leukemia (AML) are useful, but have considerable inaccuracy and imprecision. No current model includes covariates related to immune cells in the AML microenvironment. Here, an immune risk score was explored to predict the survival of patients with AML. EXPERIMENTAL DESIGN: We evaluated the predictive accuracy of several in silico algorithms for immune composition in AML based on a reference of multi-parameter flow cytometry. CIBERSORTx was chosen to enumerate immune cells from public datasets and develop an immune risk score for survival in a training cohort using least absolute shrinkage and selection operator Cox regression model. RESULTS: Six flow cytometry-validated immune cell features were informative. The model had high predictive accuracy in the training and four external validation cohorts. Subjects in the training cohort with low scores had prolonged survival compared with subjects with high scores, with 5-year survival rates of 46% versus 19% (P < 0.001). Parallel survival rates in validation cohorts-1, -2, -3, and -4 were 46% versus 6% (P < 0.001), 44% versus 18% (P = 0.041), 44% versus 24% (P = 0.004), and 62% versus 32% (P < 0.001). Gene set enrichment analysis indicated significant enrichment of immune relation pathways in the low-score cohort. In multivariable analyses, high-risk score independently predicted shorter survival with HRs of 1.45 (P = 0.005), 2.12 (P = 0.004), 2.02 (P = 0.034), 1.66 (P = 0.019), and 1.59 (P = 0.001) in the training and validation cohorts, respectively. CONCLUSIONS: Our immune risk score complements current AML prediction models.

Optimization of hepatobiliary phase delay time of Gd-EOB-DTPA-enhanced magnetic resonance imaging for identification of hepatocellular carcinoma in patients with cirrhosis of different degrees of severity.

AIM: To optimize the hepatobiliary phase delay time (HBP-DT) of Gd-EOB-DTPA-enhanced magnetic resonance imaging (GED-MRI) for more efficient identification of hepatocellular carcinoma (HCC) occurring in different degrees of cirrhosis assessed by Child-Pugh (CP) score. METHODS: The liver parenchyma signal intensity (LPSI), the liver parenchyma (LP)/HCC signal ratios, and the visibility of HCC at HBP-DT of 5, 10, 15, 20, and 25 min (i.e., DT-5, DT-10, DT-15, DT-20, and DT-25 ) after injection of Gd-EOB-DTPA were collected and analyzed in 73 patients with cirrhosis of different degrees of severity (including 42 patients suffering from HCC) and 18 healthy adult controls. RESULTS: The LPSI increased with HBP-DT more significantly in the healthy group than in the cirrhosis group (F = 17.361, P < 0.001). The LP/HCC signal ratios had a significant difference (F = 12.453, P < 0.001) among various HBP-DT points, as well as between CP-A and CP-B/C subgroups (F = 9.761, P < 0.001). The constituent ratios of HCC foci identified as obvious hypointensity (+++), moderate hypointensity (++), and mild hypointensity or isointensity (+/-) kept stable from DT-10 to DT-25: 90.6%, 9.4%, and 0.0% in the CP-A subgroup; 50.0%, 50.0%, and 0.0% in the CP-B subgroup; and 0.0%, 0.0%, and 100.0% in the CP-C subgroup, respectively. CONCLUSION: The severity of liver cirrhosis has significant negative influence on the HCC visualization by GED-MRI. DT-10 is more efficient and practical than other HBP-DT points to identify most of HCC foci emerging in CP-A cirrhosis, as well as in CP-B cirrhosis; but an HBP-DT of 15 min or longer seems more appropriate than DT-10 for visualization of HCC in patients with CP-C cirrhosis.

[Synergistic Killing Effects of Arsenic Trioxide Combined with Itraconazole on KG1a Cells].

OBJECTIVE: To explore the effect of arsenic trioxide combined with itraconazole on proliferation and apoptosis of KG1a cells and its potential mechanism. METHODS: The cell morphology was observed with Wrighe-Giemsa staining; cell survival rate was examined by CCK-8; and colony formation capacity was measured by methylcellulose colony formation test; the flow cytometry was used to analyse the cell apoptosis rate and cell cycle; the protein expressions of BCL-2,caspase-3,BAX,SMO,Gli1 and Gli2 were detected by Western-blot. RESULTS: The arsenic trioxide and itraconazole alone both could inhibit the KG1a cell proliferation in dose-and time-dependent manner. In comparison between single and combined drug-treatment group, both the cell survival rate and the colony number of the single drug-treatment group were significantly lower(P<0.05), and the apoptosis rate was higher in the combined drug-treatment group. In the combined-treatment group, the protein expression of Caspase-3 and BAX was upregulated, while the protein expression of BCL-2,SMO,Gli1 and Gli2 was downregulated. CONCLUSION: Arsenic trioxide combined with itraconazole can inhibit the KG1a cell proliferation and induce apoptosis, which may be related with the inhibition of Hh signaling pathway and upregulation of both Caspase-3 and BAX protein expression, and provided experimental data of arsenic trioxide combined with itraconazole for the treatment of refractory AML.

Augmented PMMA distribution: improvement of mechanical property and reduction of leakage rate of a fenestrated pedicle screw with diameter-tapered perforations.

OBJECTIVE This study investigated the optimum injection volume of polymethylmethacrylate (PMMA) to augment a novel fenestrated pedicle screw (FPS) with diameter-tapered perforations in the osteoporotic vertebral body, and how the distribution characteristics of PMMA affect the biomechanical performance of this screw. METHODS Two types of FPSs were designed (FPS-A, composed of 6 perforations with an equal diameter of 1.2 mm; and FPS-B, composed of 6 perforations each with a tapered diameter of 1.5 mm, 1.2 mm, and 0.9 mm from tip to head. Each of 28 human cadaveric osteoporotic vertebrae were randomly assigned to 1 of 7 groups: FPS-A1.0: FPS-A+1.0 ml PMMA; FPS-A1.5: FPS-A+1.5 ml PMMA; FPS-A2.0: FPS-A+2.0 ml PMMA; FPS-B1.0: FPS-B+1.0 ml PMMA; FPS-B1.5: FPS-B+1.5 ml PMMA; FPS-B2.0: FPS-B+2.0 ml PMMA; and conventional pedicle screws (CPSs) without PMMA. After the augmentation, 3D CT was performed to assess the cement distribution characteristics and the cement leakage rate. Axial pullout tests were performed to compare the maximum pullout force thereafter. RESULTS The CT construction images showed that PMMA bone cement formed a conical mass around FPS-A and a cylindrical mass around FPS-B. When the injection volume was increased from 1.0 ml to 2.0 ml, the distribution region of the PMMA cement was enlarged, the PMMA was distributed more posteriorly, and the risk of leakage was increased. When the injection volume reached 2.0 ml, the risk of cement leakage was lower for screws having diameter-tapered perforations. The pullout strengths of the augmented FPS-A groups and FPS-B groups were higher than that of the CPS group (p < 0.0001). All FPS-B groups had a higher pullout strength than the FPS-A groups. CONCLUSIONS The diameter of the perforations affects the distribution of PMMA cement. The diameter-tapered design enabled PMMA to form larger bone-PMMA interfaces and achieve a relatively higher pullout strength, although statistical significance was not reached. Study results indicated 1.5-ml of PMMA was a conservative volume for PMMA augmentation; more cement injection would significantly increase the risk of cement leakage.

A Comparison of Brain Death Criteria between China and the United States.

BACKGROUND: Criteria for determining brain death (BD) vary between China and the United States. We reported the results of an investigation designed to compare procedures to determine BD in two countries. METHODS: The latest criteria in the United states were published in 2010. The latest criteria in China were published in 2009. We used these two types of BD criteria to evaluate patients who were considered to be BD. The time, cost, and accuracy of the diagnosis were compared. RESULTS: From January 1, 2012 to October 8, 2013, there were 37 patients which were applied for BD evaluation in the Neurological Intensive Care Unit of Beijing Tiantan Hospital. The cause of coma were known as subarachnoid hemorrhage (18 patients, 48.6%), intracerebral hemorrhage (8 patients, 21.6%), cerebral ischemia (9 patients, 24.3%), brain stem tumor (1 patient, 2.7%), and intracranial infection (1 patient, 2.7%). The clinical examinations were done for all of the patients except 1 patient who had low blood pressure. Three patients had brainstem reflexes that were excluded from BD. Twenty-five patients had apnea tests, and 20 tests were completed that were all positive. Confirmatory tests were completed differently: Transcranial Doppler (30 patients, positive rate 86.7%), electroencephalogram (25 patients, positive rate 100%), and somatosensory evoked potential (16 patients, positive rate 100%). Thirty-three patients were diagnosed BD by criteria of the United States. Only 9 patients were diagnosed BD by Chinese criteria. The use of time and money in the USA criteria was obviously fewer than those in Chinese criteria (P = 0.000). CONCLUSION: Compared with BD criteria of the United States, Chinese criteria were stricter, lower positive rate, more cost in money and time, and more reliable by families and doctors.

[Case control study on therapeutic effects of dynamic external fixtor combined with limited internal fixation and cross K-wires fixation for the treatment of Pilon fractures of the proximal interphalangeal joint].

OBJECTIVE: To compare the clinical effects and safety of dynamic external fixtor combined with limited internal fixation and cross K-wires fixation for the treatment of close Pilon fractures of the proximal interphalangeal joint. METHODS: From June 2012 to June 2014, totally 41 patients (45 fingers) with close interphalangeal joint Pilon fracture were treated by dynamic external fixtor combined with limited internal fixation or cross K-wires fixation, and all the patients were followed up. In the dynamic external fixtor combined with limited internal fixation group (group A), there were 21 patients with 22 fingers, including 12 males and 9 females, with an average of (30.6±5.6) years old. In the cross K-wires fixation group (group B), there were 20 patients with 23 fingers, including 11 males and 9 females, with an average of (30.1±5.3) years old. Regular re-examination of X-ray was performed to evaluate the active range of joint motion, fracture healing time, infection rate and postoperative joint motion pain. RESULTS: According to the evaluation criteria of upper extremity function issued by the Hand Surgery Society of Chinese Medical Association, the excellent and good cases of group A was up to 19 and 13 for group B. The evaluation results has significant differences (Z=2.558, P=0.011). The excellent and good rate of group A was obviously higher than that of group B. The average bone union time of group A was (7.9±2.1) weeks, and (8.1±2.3) weeks for group B. There was no significant difference on the mean healing time (t=-0.304, P=0.762). The infection fingers of group A was 5, and 1 for group B. The difference between the results was statistically significant (χ2=3.287, P<0.05). The infection rate of group A was higher than that of group B. The postoperative joint motion pain was evaluated by VAS score, the mean score was 0.18±0.50 in group A, and 0.65±0.88 in group B. The difference between the results was statistically significant (t=-2.207, P<0.05). The postoperative joint motion pain was lower than that of group B. CONCLUSION: Dynamic external fixtor combined with limited internal fixation is a reliable and effective method to treat Pilon fractures of the proximal interphalangeal joint. It allows early postoperative functional rehabilitation and restores the joint function.

[Effect of honokiol on proliferation and apoptosis in HL-60 cells and its potential mechanism].

This study was aimed to investigate the effect of Honokiol (HNK) on proliferation and apoptosis of acute myeloid leukemia HL-60 cells and its potential mechanism. Inhibitory effect of HNK on the HL-60 cell proliferation was detected by MTT assay. Flow cytometry was used to detect the change of cell cycle and AnnexinV/PI staining was used to detect apoptosis. Western blot was applied to analyze the cell cycle protein (cyclins), cyclin-dependent kinase (CDK), P53, P21, P27, BCL-2, BCL-XL, Bax, caspase-3/9 and proteins for MAPK signal pathway. The results showed that HNK could inhibit the proliferation of HL-60 cells in time- and dose dependent ways. HNK arrested HL-60 cells in G0/G1 phase, and S phase cells decreased significantly (P < 0.05). The expression of cyclin D1, cyclin A, cyclin E and CDK2/4/6 were significantly down-regulated (P < 0.05), the expression of P53 and P21 was significantly upregulated after treating for 24 h with HNK (P < 0.05). After 24 h treatment with HNK, HL-60 cell apoptosis increased significantly with the upregulation of activated caspase-3, -9, BAX expression and the downregulation of BCL-2, BCL-XL expression. The MAPK subfamily, P38 and JNK were not significantly changed, but the expression of MEK1/2-ERK1/2 was significantly downregulated (P < 0.05). It is concluded that HNK arrestes the cells at G0/G1 phase and induces HL-60 cell apoptosis through the intervention of MEK1/2-ERK1/2 signaling pathway.

[Synergistic killing effect of arsenic trioxide combined with curcumin on KG1a cells].

This study was aimed to explore the effect of arsenic trioxide combined with curcumin on proliferation and apoptosis of KG1a cells and its potential mechanism. The cell survival rate was mesured by MTT; colony formation capacity was examined by methylcellulose colony formation test; flow cytometry was used to analyse the cell surface molecules, cell apoptosis rate and cell cycle; the cell morphology was observed with Wright-Giemsa staining and the protein expression of BCL-2, BAX, PARP was detected by Western blot. The results showed that the phenotype of KG1a cells was CD34(+)CD38(-), while the phenotype of HL-60 cell was CD34(+)CD38(+). The former possessed a stronger colony ability than the latter. Effect of curcumin and arsenic trioxide alone on cell proliferation and inhibition was in dose-dependent manner. Compared with single drug-treatment group, the cell survival rate and colony number were lower, and the apoptosis rate was higher in combined drug-treatment group. Protein expression of BCL-2 and PARP was upregulated, while the protein expression of PARP was downregulated in the combined treatment group. It is concluded that compared with HL-60 cells, KG1a cells are the earlier leukemia stem/progenitor cells. Arsenic trioxide combined with curcumin can effectively inhibit the KG1a cell proliferation and induce apoptosis, which may be associated with the downregulation of BCL-2 and PARP protein expression and the upregulation of BAX protein expression.

Verapamil reverses cardiac iron overload in streptozocin-induced diabetic rats.

Accumulating evidence shows that iron overload is a new risk factor for diabetes mellitus. L-type Ca(2+) channel (LTCC) has been identified as an important mediator for ferrous iron uptake into cardiomyocytes. In this study, we aimed to examine the effects of verapamil, the LTCC blocker, on myocardial iron metabolism in diabetic rats. Diabetes was induced by intraperitoneal injection of streptozocin after intragastric administration of fat emulsion, and then treated by verapamil (5 mg · kg(-1) · day(-1)) for 1 week. The results showed that verapamil did not alter the blood glucose level of diabetic rats. However, elevated levels of superoxide dismutase, malonaldehyde, and serum ferritin in diabetic rats were decreased significantly by verapamil treatment. Moreover, serum, myocardial, and urine iron were elevated remarkably along with a decrease of hepatic iron in diabetic rats. After verapamil administration, serum and myocardial iron in diabetic rats were reduced significantly but urine and hepatic iron were increased. Furthermore, confocal microscopy demonstrated that intracellular-free iron concentration was elevated dramatically in cardiomyocytes of diabetic rats, which was markedly attenuated after verapamil treatment. In summary, our data demonstrated that verapamil prevented myocardial iron overload by inhibiting intracellular iron accumulation in diabetic cardiomyocytes.

[Effects of limb ischemia preconditioning on pulmonary free radicals and cytokine levels in a rabbit model of lung ischemia-reperfusion injury].

OBJECTIVE: To study the effects of limb ischemia preconditioning on pulmonary free radicals and cytokine levels during lung ischemia-reperfusion injury in rabbits. METHODS: Eighteen healthy rabbits were randomly divided into three groups: control group (group C, n = 6), ischemia/reperfusion group (group I/R, n = 6), limb ischemia preconditioning group (group L, n = 6). At the end of experiments, the wet to dry-weight ratio (W/D), activities of superoxide dismutase (SOD) and myeloperoxidase (MPO), levels of malondialdehyde (MDA) and the contents of cytokines (TNF-α, IL-6, IL-8 and IL-10) were determined in lung tissues. Protein levels of bronchoalveolar lavage fluid and serum were measured to calculate the lung permeability index. Pathologic changes of lung tissues were also observed. RESULTS: Compared to the group I/R, the lung tissue W/D ratio, MPO activity, lung permeability index, MDA and the cytokines (TNF-α, IL-6 and IL-8) levels were significantly decreased in group L (P < 0.05), while the SOD activity (P < 0.05) and IL-10 contents were significantly increased (P < 0.01). There was no statistical difference in the changes of the above parameters between group L and group C (P > 0.05). The morphologic damages were significantly reduced in group L than that in group I/R. CONCLUSION: Limb ischemia preconditioning has protective effect against lung ischemia-reperfusion injury, which may at least in part through inhibiting the release of oxygen-derived free radicals and pro-inflammatory cytokines (TNF-α, IL-6, IL-8) and increasing the production of anti-inflammatory cytokine IL-10.

[Study on immunological effect of housefly protein in mice].

OBJECTIVE: To study the immunological effect of the housefly protein in mice. METHODS: The housefly protein was given orally in normal and Cyclophosphamide (CY) immunosuppressive mice, and then the indexes of the clearance rate of carbon particles and the serum hemolysin against SRBC were detected. RESULTS: The housefly protein could increase HC50 significantly in the normal mice, but could not in the CY immunosppressive mice. It could increase phagocytic index significantly in the normal mice, but could not in the CY immunosuppressive mice. CONCLUSION: The results suggest that the housefly protein can promote humoral immunity functions of normal mice. It has distinct preventive and cure effect on the CY immunosuppressive.

[Isolation and purification of antibacterial peptides from the larvae secretion of housefly and the characteristics].

OBJECTIVE: To isolate and purify the antibacterial peptides from larvae secretion of housefly (Musca domestica) and study their partial characteristics. METHODS: Protein isolation and purification were performed by routine process, namely, ultrafiltration, solid phase extraction (SPE) and reversed-phase high-performance liquid chromatography (RP-HPLC). The minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC) of the antibacterial peptides were examined. The antibacterial effect of peptides was studied in nutritive medium with different pH value(pH 5.0-10.0), divalent cations (Mg2+: (0.5 x 10(-3) - 10.0 x 10(-3))mol/L, Na+, K+: (10 x 10(-3) - 100 x 10(-3))mol/L), and serum content (12.5%-75%). RESULTS: Molecular weight of the peptides was about Mr 3 000-30 000 after ultrafiltration. The fractions eluted with 20%, 30%, 70%, and 80% of acetonitrile (ACN) all showed antibacterial activity by solid phase extraction. The fractions eluted with 70% ACN showed strongest and stable st antibacterial activity which was further purified by RP-HPLC. Two sub-fractions appeared at around RT 15.5 min and 18.5 min were obtained with antibacterial activity. The MIC to those standard Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus, and Bacillus subtilis was 32.7380, 16.3688, 65.4750 and 32.7380 microg/ml respectively. In the nutritive medium of pH 6.0-9.0, different divalent cations and serum content, the increment of A570 in experiment groups was less than 0.05, while that of the control group was greater than 0.3 (P<0.01). CONCLUSION: SPE and RP-HPLC have been effective in purifying the antibacterial peptides which show strong activity in different conditions.